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The Role of Circulating Tumor Cells in the Prognosis of Local Recurrence and Local Residual Nasopharyngeal Carcinoma Undergoing Endoscopic Resection

The Role of Circulating Tumor Cells in the Prognosis of Local Recurrence and Local Residual... Hindawi Journal of Oncology Volume 2022, Article ID 1453792, 8 pages https://doi.org/10.1155/2022/1453792 Research Article The Role of Circulating Tumor Cells in the Prognosis of Local Recurrence and Local Residual Nasopharyngeal Carcinoma Undergoing Endoscopic Resection 1,2,3 4 5 Liang Tang, Zhifeng Guan , and Mengdi Xu Department of Otorhinolaryngology, Minzu Hospital of Guangxi Zhuang Autonomous Region, Guangxi, China Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor (Guangxi Medical University), Ministry of Education, Guangxi, China Guangxi Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor, Guangxi, China Department of Radiotherapy, Tumor Hospital Aˆliated to Nantong University, Nantong Tumor Hospital, Nantong 226000, China Zhejiang Baiyining Medical Laboratory Co,.Ltd, Zhejiang, China Correspondence should be addressed to Zhifeng Guan; 5300671@ntu.edu.cn Received 14 July 2022; Revised 5 August 2022; Accepted 13 August 2022; Published 12 September 2022 Academic Editor: Dong-Hua Yang Copyright © 2022 Liang Tang et al. „is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Purpose. To investigate the role of circulating tumor cells in the prognosis of local recurrence and local residual nasopharyngeal carcinoma undergoing endoscopic surgery. Methods. A total of 56 patients with locally residual nasopharyngeal carcinoma (NPC) who underwent nasal endoscopic surgery from August 2018 to December 2021 were included. „e status of circulating tumor cells (CTC) before and after surgery was detected, and its relationship with clinical characteristics and postoperative survival was analyzed. Results. After nasal endoscopy, the positive rates of CTC and mesenchymal CTC (MCTC) detected in patients with nasopharyngeal carcinoma were signi“cantly lower than those before treatment (P  0.0376; P  0.0212). Before nasal endoscopy, the status of CTC and MCTC was signi“cantly correlated with the Tstage (P < 0.05). After nasal endoscopy, the status of CTC and MCTC was signi“cantly correlated with the TNM stage, T stage, and “rst radiotherapy mode (P < 0.05). „e PFS of patients with di—erent clinical characteristics was analyzed, and the results showed that the PFS of NPC patients with CTC (+) was signi“cantly shorter than that of CTC (−) patients (18.71 vs. 22.47, P < 0.05) and the PFS of NPC patients with MCTC (+) was signi“cantly shorter than that of MCTC (−) patients (18.22 vs. 22.30, P < 0.05). „e PFS of NPC patients in TNM stage (I-II) was signi“cantly longer than that in TNM stage (III) patients (22.53 vs. 18.57, P < 0.05). „e PFS of NPC patients whose “rst radiotherapy mode was conventional was signi“cantly longer than that of patients whose “rst radiotherapy mode was enhanced (22.14 vs. 16.85, P < 0.05). „e COX analysis showed that MCTC and TNM stages were independent risk factors a—ecting the prognosis of local recurrence or local residual nasopharyngeal carcinoma after endoscopic resection (P < 0.05). Conclusion. „e detection of CTC is helpful for the prognosis evaluation of local recurrence or local residual NPC after endoscopic resection of NPC. „e MCTC is an important factor a—ecting the prognosis of NPC patients. radiotherapy. A recent study reported that 14% of 3328 NPC 1. Introduction patients who received radiotherapy as the primary treatment Nasopharyngeal carcinoma (NPC) is one of the most modality had local residual or local recurrence [2]. In recent common malignant tumors in China. „e current treatment years, with the deepening research on endoscopic skull base for NPC is radiotherapy, with or without chemotherapy. anatomy, the increasingly sophisticated endoscopic equip- „is treatment mode has achieved ideal ežcacy [1]. How- ment and instruments, and the sophisticated surgical ever, there are cases with local residual tumor cells after techniques, transnasal endoscopic resection has become an 2 Journal of Oncology important alternative option for patients with locally re- residual NPC in an anticoagulation tube, it was shaken sidual or locally recurring NPC [3]. However, transnasal gently to prevent blood coagulation. Blood samples were endoscopic is a local treatment, and a comprehensive collected one week before surgery and one month after treatment (radiotherapy or chemotherapy) using various surgery. /e sample was centrifuged (2500 r/min 5 min) to methods is still required after surgery according to different remove the supernatant and was fixed for 8 min. After histological types and clinical stages to achieve the best centrifugation, the pellet was mixed in 100 ml of 10∗ PBS for curative effect. /erefore, the prognostic evaluation of 10 seconds. . /en, 900 ml of water was added and centri- transnasal endoscopic resection is very important for clinical fuged at 1000 rpm for 5 min until no red blood cells are seen decisions. in the pellet. /e filter was connected to a vacuum pump Circulating tumor cells (CTCs) are the general term for through a vacuum manifold, and the liquid in the sample various types of tumor cells shed from solid tumors into the was transferred to the filter and filtrated through an 8 μm peripheral circulation [4]. It is believed that CD45−/ pore size filter. CTCs and some leukocytes are trapped in the EpCAM+/CK+ cells in peripheral blood are CTCs, and their filter. /e filtration membrane was fixed with formaldehyde status is closely related to the clinical stage, lymph node solution at room temperature for 60 min. metastasis, degree of differentiation, and prognosis of tumor patients and plays an important role in tumor recurrence, 3.2. Incubation and Staining. /e samples were incubated metastasis, and prognosis evaluation [5]. Some studies have with a permeabilizer for 5 min and washed three times with indicated that the progression-free survival and overall PBS solution 0.1 × SSC (Sigma, St. Louis, USA). /e samples survival of cancer patients with CTCs in peripheral blood are were incubated with digestive enzymes for 60 min and significantly shortened [6]. At present, CTC has been used in washed three times with PBS. After adding the probe the prognostic evaluation of various tumors [7, 8], but it is working solution (specific capture probes: epithelial bio- rarely used to evaluate the prognosis of patients with locally marker probes EpCAM, CK8/18/19, mesenchymal bio- residual or locally recurrent NPC in patients undergoing marker probes vimentin and twist, and leukocyte marker endoscopic resection. /is study evaluated the role of CTC CD45), the sample was placed in a 40 C biochemical in- in the prognosis of NPC. cubator for 3 h and washed three times with PBS. /en, the preamplification working solution (30% horse serum, 1.5% 2. Materials and Methods sodium dodecyl sulfate, 3 mMI Tris-HCl (pH 8.0), and 0.5 fmol preamplification probe) was added to the sample 2.1. Research Subject. A total of 56 patients with local re- and incubated for 30 min in a 40 C biochemical incubator. currence and local residual NPC who underwent nasal /e sample was washed with PBS three times before adding endoscopic treatment in Nantong Tumor Hospital from amplification working solution (30% horse serum, 1.5% August 2018 to December 2021 were included. Inclusion sodium dodecyl sulfate, 3 mMI Tris-HCl (pH 8.0), and criteria were as follows: (1) Patients with locally recurrent 1 fmol amplification probe) and placed in a 40 C bio- NPC, which is defined by local tumor recurrence observed chemical incubator for 30 min and again washed three times by nasal endoscopy after 6 months of radiotherapy. Tissues with PBS. were collected and confirmed by pathological examination. We used a fluorophore-labeled probe to hybridize with (2) Patients with local residual disease, which was defined by the amplification probe to generate a fluorescent signal. Add a residual mass in the nasopharynx and/or parapharyngeal the fluorescent dyes Alexa Fluor 488 (interstitial biomarker space, were found by nasal endoscopy and MRI after probe vimentin and twist) and Alexa Fluor 750 (for labeling 3 months of radiotherapy. Tumor tissue was collected and leukocytes) (Marker CD45) to the sample and incubate for confirmed by pathological examination. (3) Patients were 30 min in a 40 C biochemical incubator. /e sample was treated with radiotherapy for nasopharyngeal carcinoma. washed three times with PBS before Antiquencher (con- Reradiotherapy in the short term may cause serious com- taining DAPI for nuclear fluorescent staining) was added to plications such as bone necrosis, radiation-induced brain the sample. /e results were observed directly after the cell damage, internal carotid artery rupture, and hemor- samples were placed for 5 min. rhage, and they cannot continue to receive radiotherapy. (4) /e patient’s general condition is good and can tolerate general anesthesia surgery. Exclusion criteria were as fol- 3.3. Interpretation of Results. After hybridization and lows: (1) Patients with intracranial metastatic lesions staining with different specific probes, different fluorescent revealed by CT or MRI examinations of the cranial brain, signals can be identified. Interstitial CTC (MCTC) is a type nasopharynx, and paranasal sinuses. (2) Patients with distant of CTC. /is study mainly explored the status of CTC and metastases. (3) Patients with surgical contraindications, such MCTC. /e peripheral blood CTC≥ 3/5 ml of patients with as patients with severe liver, kidney, cardiac insufficiency, nasopharyngeal carcinoma was judged as positive, and <3/ and severe coagulation dysfunction. 5 ml was judged as CTC negative. MCTC≥ 1/5 ml was judged as positive, and <1/5 ml was judged as CTC negative. 3. Detection of CTCs in Peripheral Blood 3.1.EnrichmentofCTCs. After collecting 15 ml of peripheral 3.4. Surgical Procedure. We kept the patient in a supine venous blood from patients with local recurrence or local position, inserted a special trachea into the patient’s trachea Journal of Oncology 3 through the mouth, and performed general anesthesia. /en, gender, N staging, M staging, and the tumor state (P> 0.05) we used cotton piece to take 0.1% of adrenalin to make nasal (Tables 1 and 2). mucosa of the patient fully contracted. /e mucosa 5 mm away from the residual lesion is cut under the nasal en- 4.3. PFS Analysis. During the follow-up period, no patients doscope using a high-frequency electric knife or a low- died. /e disease progressed in 16 patients. Five patients had temperature plasma knife until they reach the bone. /e a second recurrence in the nasopharynx. Six patients had mucosa and the bone surface are separated with a nerve a first recurrence in the nasopharynx. Primary second tu- dissection stick close to the incision, thereby excising the mors developed in 5 patients, 3 of the tongue and 2 of the lesions. After the operation, the nostrils were stuffed and palate. /e PFS of patients with different clinical charac- washed using a large amount of distilled water to completely teristics was analyzed, and the results showed that the PFS of clean the cavity and compress the bleeding. When special NPC patients with CTC (+) was significantly shorter than circumstances are encountered during the operation, such as that of CTC (−) patients (18.71 vs. 22.47, P< 0.05). /e PFS the soft tissue of the nasopharynx cannot be cut at one time, of NPC patients with MCTC (+) was significantly shorter the method of partial excision is adopted. Meanwhile, the than that of MCTC (−) patients (18.22 vs. 22.30, P< 0.05). tissue 1 cm behind the turbinate around the malignant tu- /e PFS of NPC patients in TNM stage (I-II) was signifi- mor was treated with hemostasis. Postoperative adjuvant cantly longer than that in TNM stage (III) patients (22.53 vs. chemotherapy was given to the patient. Adjuvant chemo- 18.57, P< 0.05). /e PFS of NPC patients was significantly therapy was a TP regimen: docetaxel 75 mg/m , d1. Cis- 2 longer in patients whose first radiotherapy mode was platin25 mg/m , d1–3. One cycle lasted for 21 days, and the conventional than in patients whose first radiotherapy mode patient received chemotherapy for 2-3 consecutive cycles. was enhanced (22.14 vs. 16.85, P< 0.05). /ere was no significant difference in PFS between the CTC (+) group, the 3.5. Postoperative Follow-Up. All patients were followed up CTC (−) group, the MCTC (+) group, and the MCTC (−) by telephone after endoscopic nasal resection. Follow-up group D (P> 0.05) (Tables 3 and 4, Figure 2). was terminated when the patient’s disease progressed, died, or did not respond at the last follow-up. /e end of follow-up 4.4. COX Univariate and Multivariate Analyses. in this study was January 1, 2022. /e median follow-up time According to COX analysis, MCTC and TNM staging were was 20 months. PFS is the time from the first day after independent risk factors affecting the prognosis of local surgery to the patient’s first disease progression or death recurrence or local residual nasopharyngeal carcinoma after from any cause. /e interval between recurrences was the endoscopic resection (P< 0.05) (Tables 5 and 6). time from the end of radiotherapy to the time when re- currences were confirmed by examination. 5. Discussion 4. Results For NPC patients with residual or locally recurrent lesions, given the high incidence of severe toxicity associated with 4.1. Changes in CTC Status before and after Treatment. reirradiation, surgery should be considered for locally re- Before undergoing transnasal endoscopic resection, 34 out curring and resectable cases. Transnasal endoscopic surgical of 56 patients (60.71%) had positive CTCs with a median resection is often used clinically. Compared with traditional count of 6.00 CTCs (range, 0–58 per 5 mL of blood). 29 surgical methods, transnasal endoscopic resection results in (51.79%) patients were positive for MCTCs with a median less trauma, less damage to surrounding tissues, and less count of 5.00 CTCs (range, 0–64 per 5 mL of blood). At bleeding. In addition, transnasal endoscopic resection has 1 month after transnasal endoscopy, CTCs were detected in a clearer vision, a thin mirror body, and lesions can be 23 (41.07%) patients with a median count of 5.00 CTCs observed from multiple angles, which improves the com- (range, 0–48 per 5 mL of blood); and 17 (51.79%) patients plete removal of the tumor [9]. After resection, patients with were positive for MCTCs, with a median count of 3.00 CTCs NPC still need to receive reradiotherapy or adjuvant che- (range, 0–39 per 5 mL of blood). After nasal endoscopy, the motherapy to consolidate the curative effect. /erefore, positive rates of CTC and MCTC detected in patients with a reasonable evaluation of the postoperative prognosis of nasopharyngeal carcinoma were significantly lower than NPC patients is crucial for determining the follow-up those before treatment (c2 � 4.323, P � 0.0376; c2 � 5.312, treatment [10]. TNM staging is considered to be the most P � 0.0212) (Figure 1). valuable prognostic factor affecting the treatment of naso- pharyngeal carcinoma. However, TNM staging only con- 4.2. CTC Status and Clinical Features. Before nasal endos- siders the anatomical location of the tumor and does not take copy, the status of CTC and MCTC was significantly cor- into account the heterogeneity of the tumor. /us, in clinical related with T stage (P< 0.05), but had no significant practice, the same treatment might lead to a completely correlation with age, gender, TNM stage, N stage, M stage, different effect on patients with the same TNM staging. tumor state, and the first course of radiotherapy (P> 0.05). /erefore, it is of great value to screen out economic, ob- After nasal endoscopy, the status of CTC and MCTC was jective, and easily detectable indicators to supplement TNM significantly correlated with TNM staging, Tstaging, and the staging to predict the prognosis of patients with nasopha- first course of radiotherapy (P< 0.05), but not with age, ryngeal carcinoma. 4 Journal of Oncology 80 80 P=0.0212 P=0.0376 60 60 40 40 20 20 0 0 Before treatment After treatment Before treatment After treatment CTC (-) CTC (-) CTC (+) CTC (+) (a) (b) Figure 1: Changes in CTC expression before and after treatment. Table 1: Correlation between preoperative CTC status and clinical characteristics. CTC MCTC Items P P N (n  22) P (n  34) N (n  27) P (n  29) Age ≤60 12 20 0.7520 14 18 0.4401 >60 10 14 13 11 Gender Male 14 23 0.7569 17 20 0.6355 Female 8 11 10 9 TNM stage I-II 12 18 0.9064 15 11 0.1864 III 10 16 12 18 T stage (in“ltration depth) T1-T2 15 12 0.0162 20 7 0.0002 T3-T4 7 22 7 22 N stage (lymphatic metastasis) N0-1 12 21 0.5917 17 16 0.5538 N2 10 13 10 13 Tumor state Local recurrence 12 21 0.8876 15 18 0.6206 Local remnants 10 13 12 11 First course of radiotherapy Normal 17 19 0.1028 20 16 0.1402 Adjust the strong 5 15 7 13 Time between recurrence (years) ≤2 4 8 0.7844 5 7 0.7411 >2 8 13 10 11 Note: N means negative. P means positive. CTC detection has become the representative of the stronger migration and invasion ability than other pheno- emerging liquid biopsy nowadays. Compared with tradi- types of CTC, which is also closely related to the clinical tional histopathological biopsy, CTC detection has the ad- characteristics of tumor patients. Si et al. [11] reported that vantages of convenience, noninvasiveness, and real-time CTC levels were closely related to NPC staging, and total results and has been widely used in the diagnosis of various CTC counts were signi“cantly positively correlated with tumor diseases and the evaluation of patient prognosis. NPC clinical staging. „e results of this study also showed CTCs are derived from clones at the primary tumor site that the status of CTC and MCTC was signi“cantly corre- and have various cellular phenotypes, including epithelial lated with T staging both before and after surgery (P < 0.05). CTCs, mixed epithelial and mesenchymal CTCs, and In addition, the status of CTC and MCTC after operation was also signi“cantly correlated with TNM staging and the mesenchymal CTCs (MCTCs). Cell phenotype can re¥ect the malignancy of tumor cells, among which MCTC has “rst radiotherapy pattern, indicating that CTC detection is Number of cases (n) Number of cases (n) Journal of Oncology 5 Table 2: Correlation between postoperative CTC status and clinical characteristics. CTC MCTC Items P P N (n � 33) P (n � 23) N (n � 39) P (n � 17) Age ≤60 16 16 0.1168 21 11 0.4502 >60 17 7 18 6 Gender Male 23 14 0.4925 26 11 0.8867 Female 10 9 13 6 TNM stage I-II 20 3 0.0004 22 1 0.0004 III 13 20 17 16 T stage (infiltration depth) T1-T2 20 7 0.0262 23 4 0.0147 T3-T4 13 16 16 13 N stage (lymphatic metastasis) N0-1 19 14 0.8053 23 10 0.9916 N2 14 9 16 7 Tumor state Local recurrence 18 15 0.4245 21 12 0.2416 Local remnants 15 8 18 5 First course of radiotherapy Normal 25 11 0.0319 29 7 0.0172 Adjust the strong 8 12 10 10 Time between recurrence (years) ≤2 6 4 0.6782 7 3 0.6163 >2 12 11 14 9 Note: N means negative. P means positive. Table 3: PFS analysis. Subject PFS 95% CI P CTC (+) 18.71± 1.18 16.39–21.04 0.0481 (−) 22.47± 0.75 21.01–23.93 MCTC (+) 18.22± 1.31 15.64–20.81 0.0027 (−) 22.30± 0.72 20.88–23.71 Age ≤60 19.10± 1.15 16.86–21.34 0.0782 >60 21.81± 1.01 19.82–23.79 Gender Male 20.19± 1.01 18.22–22.16 0.9345 Female 20.23± 1.37 17.54–22.92 TNM stage I-II 22.53± 0.83 20.91–24.15 0.0164 III 18.57± 1.16 16.28–20.86 T1-T2 20.99± 1.01 19.02–22.97 0.3587 T3 19.41± 1.23 17.00–21.83 N0-1 21.07± 0.94 19.23–22.91 0.2088 N2 18.98± 1.41 16.22–21.74 Tumor state Local recurrence 20.86± 0.98 18.96–22.78 0.3352 Local residual 19.34± 1.36 16.68–21.99 First course of radiotherapy Normal 22.14± 0.71 20.76–23.52 0.0026 Adjust the strong 16.85± 1.61 13.69–20.00 Time between recurrence (years) >2 21.93± 1.35 19.29–24.58 0.4460 ≤2 20.31± 1.30 17.75–22.86 6 Journal of Oncology Table 4: PFS analysis of di—erent CTCS and MCTC status. Subject PFS 95% CI P CTC (+) Local recurrence 19.56 ± 1.38 16.86–22.25 0.4841 Local residual 17.17 ± 2.07 13.12–21.22 CTC (−) Local recurrence 22.91 ± 1.04 20.86–24.96 0.2257 Local residual 21.91 ± 1.06 19.83–23.97 MCTC (+) Local recurrence 19.51 ± 1.51 16.55–22.47 0.2776 Local residual 16.03 ± 2.27 11.59–20.47 MCTC (−) Local recurrence 22.41 ± 1.04 20.37–24.46 0.5587 Local residual 22.21 ± 0.92 20.42–24.01 1.00 1.00 0.80 0.80 0.60 0.60 0.40 0.40 0.20 0.20 0.00 0.00 .00 5.00 10.00 15.00 20.00 25.00 .00 5.00 10.00 15.00 20.00 25.00 Time (months) Time (months) Group Group MCTC (+) CTC (+) CTC (+) censored MCTC (+) censored MCTC (-) CTC (-) CTC (-) censored MCTC (-) censored (a) (b) 1.00 1.00 0.80 0.80 0.60 0.60 0.40 0.40 0.20 0.20 0.00 0.00 .00 5.00 10.00 15.00 20.00 25.00 5.00 10.00 15.00 20.00 25.00 Time (months) Time (months) Group Group First course of First course of radiotherapy TNM stage I-II TNM stage I-II censored radiotherapy (normal) (normal) censored TNM stage III TNM stage III censored First course of radiotherapy First course of radiotherapy (adjust the strong) (adjust the strong) censored (c) (d) Figure 2: PFS analysis. of great signi“cance for NPC patients regardless of before with TNM staging. „erefore, expanding the sample size, and after operation. However, there is some controversy revalidating the correlation of CTC status with TNM is about the correlation between CTC and TNM staging. „e warranted in future experiments. studies of Li et al. [12, 13] clearly pointed out that the positive CTCs are also related to the prognosis of tumor patients. rate of CTC in NPC patients has no signi“cant correlation Studies have reported that CTC-positive breast cancer, Cumulative survival rate (%) Cumulative survival rate (%) Cumulative survival rate (%) Cumulative survival rate (%) Journal of Oncology 7 Table 5: COX univariate analysis. 95.00% CI Items B SE Wald Df P Exp (B) Upper Lower CTC 1.091 0.5782 3.566 1 0.0594 2.978 0.9608 9.241 MCTC 1.126 0.5363 4.408 1 0.0361 3.083 1.078 8.818 Age −0.969 0.5726 2.871 1 0.0921 0.3791 0.1245 1.164 Gender −0.042 0.5094 0.0071 1 0.9342 0.9597 0.3538 2.601 TNM stage 1.418 0.6383 4.947 1 0.0263 4.130 1.183 14.41 T 0.4501 0.4942 0.8293 1 0.3622 1.568 0.5952 4.130 N 0.6042 0.4871 1.538 1 0.2156 1.829 0.7046 4.746 Tumor state 0.4646 0.4862 0.9120 1 0.3400 1.591 0.6131 4.127 First course of radiotherapy 1.411 0.5082 7.733 1 0.0062 4.099 1.513 11.10 Table 6: COX multifactor analysis. 95.00% CI Subject B SE Wald Df P Exp (B) Upper Lower MCTC 1.673 0.7799 4.600 1 0.0319 5.327 1.155 24.57 TNM stage 1.094 0.4583 5.696 1 0.0169 2.986 1.216 7.332 First course of radiotherapy 0.675 0.5185 1.693 1 0.1932 1.963 0.7110 5.425 colorectal cancer, and liver cancer have a poor prognosis and there are many important tissues and organs around the shorter survival time [14–17]. In nasopharyngeal carcinoma, nasopharynx. Intensive radiotherapy will aggravate the Zhang [18] found that after 2–4 cycles of chemotherapy, the damage to the surrounding tissues and will also have a certain impact on the prognosis. Another reason is the CTC count of NPC was reduced remarkably, and the change in CTC number was correlated with treatment response. small number of cases in this study, the short follow-up time, and the lack of analysis of the long-term adverse reactions of Qian [19] reported that the reduction of CTC was related to the treatment effect of patients with advanced NPC, and the the patients. /erefore, whether the first radiotherapy positive CTC before treatment was an independent risk method affects the prognosis of NPC recurrence needs to be factor for poor prognosis. Yu [20] reported that CTC was further explored. /e COX analysis screened out that MCTC associated with poor survival in NPC patients, which may be and TNM staging were independent risk factors affecting the an independent prognostic factor affecting NPC. In our prognosis of local recurrence or local residual NPC after study, the positive rates of CTC and MCTC detected in endoscopic resection, suggesting that MCTC is an important patients with nasopharyngeal carcinoma after nasal en- factor affecting the prognosis of NPC patients. doscopy were significantly lower than those before treat- In conclusion, the detection of CTC is helpful for the prognosis evaluation after endoscopic nasal resection of ment, suggesting that CTC and MCTC counts changed before and after endoscopic nasal resection. For transnasal local recurrence or local residual NPC. MCTC is an im- endoscopic resection, CTC and MCTC are also potential portant factor affecting the prognosis of patients with NPC. indicators to monitor the prognosis of NPC patients. /e PFS of patients with different clinical characteristics was Data Availability analyzed, and the results showed that the PFS of CTC- positive NPC patients was significantly shorter than that /e data used to support the findings of this study are of CTC-negative patients, and the PFS of MCTC-positive available from the corresponding author upon request. NPC patients was significantly shorter than that of MCTC- negative patients. /e PFS of NPC patients in TNM stage (I- Conflicts of Interest II) was significantly longer than that of TNM stage (III) patients, and the PFS of NPC patients whose first radio- /e authors declare that they have no conflicts of interest. therapy mode was conventional was significantly longer than that of patients whose first radiotherapy mode was Acknowledgments enhanced. 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The Role of Circulating Tumor Cells in the Prognosis of Local Recurrence and Local Residual Nasopharyngeal Carcinoma Undergoing Endoscopic Resection

Journal of Oncology , Volume 2022 – Sep 12, 2022

The Role of Circulating Tumor Cells in the Prognosis of Local Recurrence and Local Residual Nasopharyngeal Carcinoma Undergoing Endoscopic Resection

Abstract

<i>Purpose</i>. To investigate the role of circulating tumor cells in the prognosis of local recurrence and local residual nasopharyngeal carcinoma undergoing endoscopic surgery. <i>Methods</i>. A total of 56 patients with locally residual nasopharyngeal carcinoma (NPC) who underwent nasal endoscopic surgery from August 2018 to December 2021 were included. The status of circulating tumor cells (CTC) before and after surgery was detected, and its relationship with clinical characteristics and postoperative survival was analyzed. <i>Results</i>. After nasal endoscopy, the positive rates of CTC and mesenchymal CTC (MCTC) detected in patients with nasopharyngeal carcinoma were significantly lower than those before treatment (<span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.2064009pt" id="M1" height="8.8423pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 8.8423"><g transform="matrix(.013,0,0,-0.013,0,0)"><path id="g113-81" d="M600 480C600 590 528 650 384 650H143L137 622C222 614 225 607 210 531L130 127C113 41 106 36 23 28L17 0H294L300 28C204 36 195 42 212 127L243 284L314 263C327 263 339 263 352 264C465 271 600 337 600 480ZM508 481C508 351 402 304 329 304C289 304 265 311 250 317L295 559C302 594 310 606 323 611C335 616 350 619 367 619C455 619 508 573 508 481Z"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><path id="g117-34" d="M535 323V373H52V323H535ZM535 138V188H52V138H535Z"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="34.445pt" style="vertical-align:-0.2064009pt" height="8.8423pt" version="1.1" viewBox="22.8711838 -8.6359 34.445 8.8423"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><path id="g113-49" d="M241 635C89 635 35 457 35 312C35 153 89 -12 240 -12C390 -12 443 166 443 312C443 466 390 635 241 635ZM238 602C329 602 354 454 354 312C354 172 330 22 240 22C152 22 124 173 124 313S148 602 238 602Z"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><path id="g113-47" d="M113 -12C146 -12 170 11 170 46C170 78 146 103 114 103S58 78 58 46C58 11 82 -12 113 -12Z"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><path id="g113-52" d="M285 378C315 398 338 416 353 432C373 451 384 474 384 503C384 579 325 635 236 635H235C182 635 136 610 108 579L65 516L85 496C110 533 150 575 205 575C258 575 300 543 300 481C300 407 232 369 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width="19.289pt" style="vertical-align:-0.2064009pt" id="M2" height="8.8423pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 8.8423"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><use xlink:href="#g117-34"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="34.445pt" style="vertical-align:-0.2064009pt" height="8.8423pt" version="1.1" viewBox="22.8711838 -8.6359 34.445 8.8423"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><path id="g113-51" d="M412 140C382 77 369 73 315 73H129L270 222C362 320 402 379 402 466C402 571 322 635 234 635C177 635 130 609 99 576L42 495L64 475C90 514 133 568 201 568C274 568 318 519 318 435C318 349 255 267 193 193C144 135 87 78 32 23V0H405C417 45 427 89 440 131L412 140Z"/></g><g transform="matrix(.013,0,0,-0.013,44.605,0)"><path id="g113-50" d="M384 0V27C293 34 287 42 287 114V635C232 613 172 594 109 583V559L157 557C201 555 205 550 205 499V114C205 42 199 34 109 27V0H384Z"/></g><g transform="matrix(.013,0,0,-0.013,50.845,0)"><use xlink:href="#g113-51"/></g></svg>).</span></span> Before nasal endoscopy, the status of CTC and MCTC was significantly correlated with the <i>T</i> stage (<span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.6370001pt" id="M3" height="9.2729pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 9.2729"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><path id="g117-91" d="M512 -3V55L134 254V256L512 456V514L75 281V230L512 -3Z"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="21.918pt" style="vertical-align:-0.6370001pt" height="9.2729pt" version="1.1" viewBox="22.8711838 -8.6359 21.918 9.2729"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><path id="g113-54" d="M153 550H386L412 615L406 623H120L82 318C104 327 142 338 184 338C294 338 347 275 347 187C347 112 305 39 221 39C160 39 119 71 97 89C88 97 80 96 71 90C59 80 50 67 49 57C48 45 52 36 66 23C80 9 123 -12 169 -12C221 -11 288 15 342 59C403 109 431 165 431 225C431 308 366 395 238 395C212 395 165 379 127 364L153 550Z"/></g></svg>).</span></span> After nasal endoscopy, the status of CTC and MCTC was significantly correlated with the TNM stage, <i>T</i> stage, and first radiotherapy mode (<span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.6370001pt" id="M4" height="9.2729pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 9.2729"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><use xlink:href="#g117-91"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="21.918pt" style="vertical-align:-0.6370001pt" height="9.2729pt" version="1.1" viewBox="22.8711838 -8.6359 21.918 9.2729"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><use xlink:href="#g113-54"/></g></svg>).</span></span> The PFS of patients with different clinical characteristics was analyzed, and the results showed that the PFS of NPC patients with CTC (+) was significantly shorter than that of CTC (−) patients (18.71 vs. 22.47, <span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.6370001pt" id="M5" height="9.2729pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 9.2729"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><use xlink:href="#g117-91"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="21.918pt" style="vertical-align:-0.6370001pt" height="9.2729pt" version="1.1" viewBox="22.8711838 -8.6359 21.918 9.2729"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><use xlink:href="#g113-54"/></g></svg>)</span></span> and the PFS of NPC patients with MCTC (+) was significantly shorter than that of MCTC (−) patients (18.22 vs. 22.30, <span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.6370001pt" id="M6" height="9.2729pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 9.2729"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><use xlink:href="#g117-91"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="21.918pt" style="vertical-align:-0.6370001pt" height="9.2729pt" version="1.1" viewBox="22.8711838 -8.6359 21.918 9.2729"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><use xlink:href="#g113-54"/></g></svg>).</span></span> The PFS of NPC patients in TNM stage (I-II) was significantly longer than that in TNM stage (III) patients (22.53 vs. 18.57, <span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.6370001pt" id="M7" height="9.2729pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 9.2729"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><use xlink:href="#g117-91"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="21.918pt" style="vertical-align:-0.6370001pt" height="9.2729pt" version="1.1" viewBox="22.8711838 -8.6359 21.918 9.2729"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><use xlink:href="#g113-54"/></g></svg>).</span></span> The PFS of NPC patients whose first radiotherapy mode was conventional was significantly longer than that of patients whose first radiotherapy mode was enhanced (22.14 vs. 16.85, <span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.6370001pt" id="M8" height="9.2729pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 9.2729"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><use xlink:href="#g117-91"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="21.918pt" style="vertical-align:-0.6370001pt" height="9.2729pt" version="1.1" viewBox="22.8711838 -8.6359 21.918 9.2729"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><use xlink:href="#g113-54"/></g></svg>).</span></span> The COX analysis showed that MCTC and TNM stages were independent risk factors affecting the prognosis of local recurrence or local residual nasopharyngeal carcinoma after endoscopic resection (<span class="inline_break"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="19.289pt" style="vertical-align:-0.6370001pt" id="M9" height="9.2729pt" version="1.1" viewBox="-0.0498162 -8.6359 19.289 9.2729"><g transform="matrix(.013,0,0,-0.013,0,0)"><use xlink:href="#g113-81"/></g><g transform="matrix(.013,0,0,-0.013,11.658,0)"><use xlink:href="#g117-91"/></g></svg><span class="irelop"/><span class="nowrap"><svg xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg" width="21.918pt" style="vertical-align:-0.6370001pt" height="9.2729pt" version="1.1" viewBox="22.8711838 -8.6359 21.918 9.2729"><g transform="matrix(.013,0,0,-0.013,22.921,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,29.161,0)"><use xlink:href="#g113-47"/></g><g transform="matrix(.013,0,0,-0.013,32.125,0)"><use xlink:href="#g113-49"/></g><g transform="matrix(.013,0,0,-0.013,38.365,0)"><use xlink:href="#g113-54"/></g></svg>).</span></span> <i>Conclusion</i>. The detection of CTC is helpful for the prognosis evaluation of local recurrence or local residual NPC after endoscopic resection of NPC. The MCTC is an important factor affecting the prognosis of NPC patients.

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References (19)

Publisher
Hindawi Publishing Corporation
ISSN
1687-8450
eISSN
1687-8469
DOI
10.1155/2022/1453792
Publisher site
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Abstract

Hindawi Journal of Oncology Volume 2022, Article ID 1453792, 8 pages https://doi.org/10.1155/2022/1453792 Research Article The Role of Circulating Tumor Cells in the Prognosis of Local Recurrence and Local Residual Nasopharyngeal Carcinoma Undergoing Endoscopic Resection 1,2,3 4 5 Liang Tang, Zhifeng Guan , and Mengdi Xu Department of Otorhinolaryngology, Minzu Hospital of Guangxi Zhuang Autonomous Region, Guangxi, China Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor (Guangxi Medical University), Ministry of Education, Guangxi, China Guangxi Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor, Guangxi, China Department of Radiotherapy, Tumor Hospital Aˆliated to Nantong University, Nantong Tumor Hospital, Nantong 226000, China Zhejiang Baiyining Medical Laboratory Co,.Ltd, Zhejiang, China Correspondence should be addressed to Zhifeng Guan; 5300671@ntu.edu.cn Received 14 July 2022; Revised 5 August 2022; Accepted 13 August 2022; Published 12 September 2022 Academic Editor: Dong-Hua Yang Copyright © 2022 Liang Tang et al. „is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Purpose. To investigate the role of circulating tumor cells in the prognosis of local recurrence and local residual nasopharyngeal carcinoma undergoing endoscopic surgery. Methods. A total of 56 patients with locally residual nasopharyngeal carcinoma (NPC) who underwent nasal endoscopic surgery from August 2018 to December 2021 were included. „e status of circulating tumor cells (CTC) before and after surgery was detected, and its relationship with clinical characteristics and postoperative survival was analyzed. Results. After nasal endoscopy, the positive rates of CTC and mesenchymal CTC (MCTC) detected in patients with nasopharyngeal carcinoma were signi“cantly lower than those before treatment (P  0.0376; P  0.0212). Before nasal endoscopy, the status of CTC and MCTC was signi“cantly correlated with the Tstage (P < 0.05). After nasal endoscopy, the status of CTC and MCTC was signi“cantly correlated with the TNM stage, T stage, and “rst radiotherapy mode (P < 0.05). „e PFS of patients with di—erent clinical characteristics was analyzed, and the results showed that the PFS of NPC patients with CTC (+) was signi“cantly shorter than that of CTC (−) patients (18.71 vs. 22.47, P < 0.05) and the PFS of NPC patients with MCTC (+) was signi“cantly shorter than that of MCTC (−) patients (18.22 vs. 22.30, P < 0.05). „e PFS of NPC patients in TNM stage (I-II) was signi“cantly longer than that in TNM stage (III) patients (22.53 vs. 18.57, P < 0.05). „e PFS of NPC patients whose “rst radiotherapy mode was conventional was signi“cantly longer than that of patients whose “rst radiotherapy mode was enhanced (22.14 vs. 16.85, P < 0.05). „e COX analysis showed that MCTC and TNM stages were independent risk factors a—ecting the prognosis of local recurrence or local residual nasopharyngeal carcinoma after endoscopic resection (P < 0.05). Conclusion. „e detection of CTC is helpful for the prognosis evaluation of local recurrence or local residual NPC after endoscopic resection of NPC. „e MCTC is an important factor a—ecting the prognosis of NPC patients. radiotherapy. A recent study reported that 14% of 3328 NPC 1. Introduction patients who received radiotherapy as the primary treatment Nasopharyngeal carcinoma (NPC) is one of the most modality had local residual or local recurrence [2]. In recent common malignant tumors in China. „e current treatment years, with the deepening research on endoscopic skull base for NPC is radiotherapy, with or without chemotherapy. anatomy, the increasingly sophisticated endoscopic equip- „is treatment mode has achieved ideal ežcacy [1]. How- ment and instruments, and the sophisticated surgical ever, there are cases with local residual tumor cells after techniques, transnasal endoscopic resection has become an 2 Journal of Oncology important alternative option for patients with locally re- residual NPC in an anticoagulation tube, it was shaken sidual or locally recurring NPC [3]. However, transnasal gently to prevent blood coagulation. Blood samples were endoscopic is a local treatment, and a comprehensive collected one week before surgery and one month after treatment (radiotherapy or chemotherapy) using various surgery. /e sample was centrifuged (2500 r/min 5 min) to methods is still required after surgery according to different remove the supernatant and was fixed for 8 min. After histological types and clinical stages to achieve the best centrifugation, the pellet was mixed in 100 ml of 10∗ PBS for curative effect. /erefore, the prognostic evaluation of 10 seconds. . /en, 900 ml of water was added and centri- transnasal endoscopic resection is very important for clinical fuged at 1000 rpm for 5 min until no red blood cells are seen decisions. in the pellet. /e filter was connected to a vacuum pump Circulating tumor cells (CTCs) are the general term for through a vacuum manifold, and the liquid in the sample various types of tumor cells shed from solid tumors into the was transferred to the filter and filtrated through an 8 μm peripheral circulation [4]. It is believed that CD45−/ pore size filter. CTCs and some leukocytes are trapped in the EpCAM+/CK+ cells in peripheral blood are CTCs, and their filter. /e filtration membrane was fixed with formaldehyde status is closely related to the clinical stage, lymph node solution at room temperature for 60 min. metastasis, degree of differentiation, and prognosis of tumor patients and plays an important role in tumor recurrence, 3.2. Incubation and Staining. /e samples were incubated metastasis, and prognosis evaluation [5]. Some studies have with a permeabilizer for 5 min and washed three times with indicated that the progression-free survival and overall PBS solution 0.1 × SSC (Sigma, St. Louis, USA). /e samples survival of cancer patients with CTCs in peripheral blood are were incubated with digestive enzymes for 60 min and significantly shortened [6]. At present, CTC has been used in washed three times with PBS. After adding the probe the prognostic evaluation of various tumors [7, 8], but it is working solution (specific capture probes: epithelial bio- rarely used to evaluate the prognosis of patients with locally marker probes EpCAM, CK8/18/19, mesenchymal bio- residual or locally recurrent NPC in patients undergoing marker probes vimentin and twist, and leukocyte marker endoscopic resection. /is study evaluated the role of CTC CD45), the sample was placed in a 40 C biochemical in- in the prognosis of NPC. cubator for 3 h and washed three times with PBS. /en, the preamplification working solution (30% horse serum, 1.5% 2. Materials and Methods sodium dodecyl sulfate, 3 mMI Tris-HCl (pH 8.0), and 0.5 fmol preamplification probe) was added to the sample 2.1. Research Subject. A total of 56 patients with local re- and incubated for 30 min in a 40 C biochemical incubator. currence and local residual NPC who underwent nasal /e sample was washed with PBS three times before adding endoscopic treatment in Nantong Tumor Hospital from amplification working solution (30% horse serum, 1.5% August 2018 to December 2021 were included. Inclusion sodium dodecyl sulfate, 3 mMI Tris-HCl (pH 8.0), and criteria were as follows: (1) Patients with locally recurrent 1 fmol amplification probe) and placed in a 40 C bio- NPC, which is defined by local tumor recurrence observed chemical incubator for 30 min and again washed three times by nasal endoscopy after 6 months of radiotherapy. Tissues with PBS. were collected and confirmed by pathological examination. We used a fluorophore-labeled probe to hybridize with (2) Patients with local residual disease, which was defined by the amplification probe to generate a fluorescent signal. Add a residual mass in the nasopharynx and/or parapharyngeal the fluorescent dyes Alexa Fluor 488 (interstitial biomarker space, were found by nasal endoscopy and MRI after probe vimentin and twist) and Alexa Fluor 750 (for labeling 3 months of radiotherapy. Tumor tissue was collected and leukocytes) (Marker CD45) to the sample and incubate for confirmed by pathological examination. (3) Patients were 30 min in a 40 C biochemical incubator. /e sample was treated with radiotherapy for nasopharyngeal carcinoma. washed three times with PBS before Antiquencher (con- Reradiotherapy in the short term may cause serious com- taining DAPI for nuclear fluorescent staining) was added to plications such as bone necrosis, radiation-induced brain the sample. /e results were observed directly after the cell damage, internal carotid artery rupture, and hemor- samples were placed for 5 min. rhage, and they cannot continue to receive radiotherapy. (4) /e patient’s general condition is good and can tolerate general anesthesia surgery. Exclusion criteria were as fol- 3.3. Interpretation of Results. After hybridization and lows: (1) Patients with intracranial metastatic lesions staining with different specific probes, different fluorescent revealed by CT or MRI examinations of the cranial brain, signals can be identified. Interstitial CTC (MCTC) is a type nasopharynx, and paranasal sinuses. (2) Patients with distant of CTC. /is study mainly explored the status of CTC and metastases. (3) Patients with surgical contraindications, such MCTC. /e peripheral blood CTC≥ 3/5 ml of patients with as patients with severe liver, kidney, cardiac insufficiency, nasopharyngeal carcinoma was judged as positive, and <3/ and severe coagulation dysfunction. 5 ml was judged as CTC negative. MCTC≥ 1/5 ml was judged as positive, and <1/5 ml was judged as CTC negative. 3. Detection of CTCs in Peripheral Blood 3.1.EnrichmentofCTCs. After collecting 15 ml of peripheral 3.4. Surgical Procedure. We kept the patient in a supine venous blood from patients with local recurrence or local position, inserted a special trachea into the patient’s trachea Journal of Oncology 3 through the mouth, and performed general anesthesia. /en, gender, N staging, M staging, and the tumor state (P> 0.05) we used cotton piece to take 0.1% of adrenalin to make nasal (Tables 1 and 2). mucosa of the patient fully contracted. /e mucosa 5 mm away from the residual lesion is cut under the nasal en- 4.3. PFS Analysis. During the follow-up period, no patients doscope using a high-frequency electric knife or a low- died. /e disease progressed in 16 patients. Five patients had temperature plasma knife until they reach the bone. /e a second recurrence in the nasopharynx. Six patients had mucosa and the bone surface are separated with a nerve a first recurrence in the nasopharynx. Primary second tu- dissection stick close to the incision, thereby excising the mors developed in 5 patients, 3 of the tongue and 2 of the lesions. After the operation, the nostrils were stuffed and palate. /e PFS of patients with different clinical charac- washed using a large amount of distilled water to completely teristics was analyzed, and the results showed that the PFS of clean the cavity and compress the bleeding. When special NPC patients with CTC (+) was significantly shorter than circumstances are encountered during the operation, such as that of CTC (−) patients (18.71 vs. 22.47, P< 0.05). /e PFS the soft tissue of the nasopharynx cannot be cut at one time, of NPC patients with MCTC (+) was significantly shorter the method of partial excision is adopted. Meanwhile, the than that of MCTC (−) patients (18.22 vs. 22.30, P< 0.05). tissue 1 cm behind the turbinate around the malignant tu- /e PFS of NPC patients in TNM stage (I-II) was signifi- mor was treated with hemostasis. Postoperative adjuvant cantly longer than that in TNM stage (III) patients (22.53 vs. chemotherapy was given to the patient. Adjuvant chemo- 18.57, P< 0.05). /e PFS of NPC patients was significantly therapy was a TP regimen: docetaxel 75 mg/m , d1. Cis- 2 longer in patients whose first radiotherapy mode was platin25 mg/m , d1–3. One cycle lasted for 21 days, and the conventional than in patients whose first radiotherapy mode patient received chemotherapy for 2-3 consecutive cycles. was enhanced (22.14 vs. 16.85, P< 0.05). /ere was no significant difference in PFS between the CTC (+) group, the 3.5. Postoperative Follow-Up. All patients were followed up CTC (−) group, the MCTC (+) group, and the MCTC (−) by telephone after endoscopic nasal resection. Follow-up group D (P> 0.05) (Tables 3 and 4, Figure 2). was terminated when the patient’s disease progressed, died, or did not respond at the last follow-up. /e end of follow-up 4.4. COX Univariate and Multivariate Analyses. in this study was January 1, 2022. /e median follow-up time According to COX analysis, MCTC and TNM staging were was 20 months. PFS is the time from the first day after independent risk factors affecting the prognosis of local surgery to the patient’s first disease progression or death recurrence or local residual nasopharyngeal carcinoma after from any cause. /e interval between recurrences was the endoscopic resection (P< 0.05) (Tables 5 and 6). time from the end of radiotherapy to the time when re- currences were confirmed by examination. 5. Discussion 4. Results For NPC patients with residual or locally recurrent lesions, given the high incidence of severe toxicity associated with 4.1. Changes in CTC Status before and after Treatment. reirradiation, surgery should be considered for locally re- Before undergoing transnasal endoscopic resection, 34 out curring and resectable cases. Transnasal endoscopic surgical of 56 patients (60.71%) had positive CTCs with a median resection is often used clinically. Compared with traditional count of 6.00 CTCs (range, 0–58 per 5 mL of blood). 29 surgical methods, transnasal endoscopic resection results in (51.79%) patients were positive for MCTCs with a median less trauma, less damage to surrounding tissues, and less count of 5.00 CTCs (range, 0–64 per 5 mL of blood). At bleeding. In addition, transnasal endoscopic resection has 1 month after transnasal endoscopy, CTCs were detected in a clearer vision, a thin mirror body, and lesions can be 23 (41.07%) patients with a median count of 5.00 CTCs observed from multiple angles, which improves the com- (range, 0–48 per 5 mL of blood); and 17 (51.79%) patients plete removal of the tumor [9]. After resection, patients with were positive for MCTCs, with a median count of 3.00 CTCs NPC still need to receive reradiotherapy or adjuvant che- (range, 0–39 per 5 mL of blood). After nasal endoscopy, the motherapy to consolidate the curative effect. /erefore, positive rates of CTC and MCTC detected in patients with a reasonable evaluation of the postoperative prognosis of nasopharyngeal carcinoma were significantly lower than NPC patients is crucial for determining the follow-up those before treatment (c2 � 4.323, P � 0.0376; c2 � 5.312, treatment [10]. TNM staging is considered to be the most P � 0.0212) (Figure 1). valuable prognostic factor affecting the treatment of naso- pharyngeal carcinoma. However, TNM staging only con- 4.2. CTC Status and Clinical Features. Before nasal endos- siders the anatomical location of the tumor and does not take copy, the status of CTC and MCTC was significantly cor- into account the heterogeneity of the tumor. /us, in clinical related with T stage (P< 0.05), but had no significant practice, the same treatment might lead to a completely correlation with age, gender, TNM stage, N stage, M stage, different effect on patients with the same TNM staging. tumor state, and the first course of radiotherapy (P> 0.05). /erefore, it is of great value to screen out economic, ob- After nasal endoscopy, the status of CTC and MCTC was jective, and easily detectable indicators to supplement TNM significantly correlated with TNM staging, Tstaging, and the staging to predict the prognosis of patients with nasopha- first course of radiotherapy (P< 0.05), but not with age, ryngeal carcinoma. 4 Journal of Oncology 80 80 P=0.0212 P=0.0376 60 60 40 40 20 20 0 0 Before treatment After treatment Before treatment After treatment CTC (-) CTC (-) CTC (+) CTC (+) (a) (b) Figure 1: Changes in CTC expression before and after treatment. Table 1: Correlation between preoperative CTC status and clinical characteristics. CTC MCTC Items P P N (n  22) P (n  34) N (n  27) P (n  29) Age ≤60 12 20 0.7520 14 18 0.4401 >60 10 14 13 11 Gender Male 14 23 0.7569 17 20 0.6355 Female 8 11 10 9 TNM stage I-II 12 18 0.9064 15 11 0.1864 III 10 16 12 18 T stage (in“ltration depth) T1-T2 15 12 0.0162 20 7 0.0002 T3-T4 7 22 7 22 N stage (lymphatic metastasis) N0-1 12 21 0.5917 17 16 0.5538 N2 10 13 10 13 Tumor state Local recurrence 12 21 0.8876 15 18 0.6206 Local remnants 10 13 12 11 First course of radiotherapy Normal 17 19 0.1028 20 16 0.1402 Adjust the strong 5 15 7 13 Time between recurrence (years) ≤2 4 8 0.7844 5 7 0.7411 >2 8 13 10 11 Note: N means negative. P means positive. CTC detection has become the representative of the stronger migration and invasion ability than other pheno- emerging liquid biopsy nowadays. Compared with tradi- types of CTC, which is also closely related to the clinical tional histopathological biopsy, CTC detection has the ad- characteristics of tumor patients. Si et al. [11] reported that vantages of convenience, noninvasiveness, and real-time CTC levels were closely related to NPC staging, and total results and has been widely used in the diagnosis of various CTC counts were signi“cantly positively correlated with tumor diseases and the evaluation of patient prognosis. NPC clinical staging. „e results of this study also showed CTCs are derived from clones at the primary tumor site that the status of CTC and MCTC was signi“cantly corre- and have various cellular phenotypes, including epithelial lated with T staging both before and after surgery (P < 0.05). CTCs, mixed epithelial and mesenchymal CTCs, and In addition, the status of CTC and MCTC after operation was also signi“cantly correlated with TNM staging and the mesenchymal CTCs (MCTCs). Cell phenotype can re¥ect the malignancy of tumor cells, among which MCTC has “rst radiotherapy pattern, indicating that CTC detection is Number of cases (n) Number of cases (n) Journal of Oncology 5 Table 2: Correlation between postoperative CTC status and clinical characteristics. CTC MCTC Items P P N (n � 33) P (n � 23) N (n � 39) P (n � 17) Age ≤60 16 16 0.1168 21 11 0.4502 >60 17 7 18 6 Gender Male 23 14 0.4925 26 11 0.8867 Female 10 9 13 6 TNM stage I-II 20 3 0.0004 22 1 0.0004 III 13 20 17 16 T stage (infiltration depth) T1-T2 20 7 0.0262 23 4 0.0147 T3-T4 13 16 16 13 N stage (lymphatic metastasis) N0-1 19 14 0.8053 23 10 0.9916 N2 14 9 16 7 Tumor state Local recurrence 18 15 0.4245 21 12 0.2416 Local remnants 15 8 18 5 First course of radiotherapy Normal 25 11 0.0319 29 7 0.0172 Adjust the strong 8 12 10 10 Time between recurrence (years) ≤2 6 4 0.6782 7 3 0.6163 >2 12 11 14 9 Note: N means negative. P means positive. Table 3: PFS analysis. Subject PFS 95% CI P CTC (+) 18.71± 1.18 16.39–21.04 0.0481 (−) 22.47± 0.75 21.01–23.93 MCTC (+) 18.22± 1.31 15.64–20.81 0.0027 (−) 22.30± 0.72 20.88–23.71 Age ≤60 19.10± 1.15 16.86–21.34 0.0782 >60 21.81± 1.01 19.82–23.79 Gender Male 20.19± 1.01 18.22–22.16 0.9345 Female 20.23± 1.37 17.54–22.92 TNM stage I-II 22.53± 0.83 20.91–24.15 0.0164 III 18.57± 1.16 16.28–20.86 T1-T2 20.99± 1.01 19.02–22.97 0.3587 T3 19.41± 1.23 17.00–21.83 N0-1 21.07± 0.94 19.23–22.91 0.2088 N2 18.98± 1.41 16.22–21.74 Tumor state Local recurrence 20.86± 0.98 18.96–22.78 0.3352 Local residual 19.34± 1.36 16.68–21.99 First course of radiotherapy Normal 22.14± 0.71 20.76–23.52 0.0026 Adjust the strong 16.85± 1.61 13.69–20.00 Time between recurrence (years) >2 21.93± 1.35 19.29–24.58 0.4460 ≤2 20.31± 1.30 17.75–22.86 6 Journal of Oncology Table 4: PFS analysis of di—erent CTCS and MCTC status. Subject PFS 95% CI P CTC (+) Local recurrence 19.56 ± 1.38 16.86–22.25 0.4841 Local residual 17.17 ± 2.07 13.12–21.22 CTC (−) Local recurrence 22.91 ± 1.04 20.86–24.96 0.2257 Local residual 21.91 ± 1.06 19.83–23.97 MCTC (+) Local recurrence 19.51 ± 1.51 16.55–22.47 0.2776 Local residual 16.03 ± 2.27 11.59–20.47 MCTC (−) Local recurrence 22.41 ± 1.04 20.37–24.46 0.5587 Local residual 22.21 ± 0.92 20.42–24.01 1.00 1.00 0.80 0.80 0.60 0.60 0.40 0.40 0.20 0.20 0.00 0.00 .00 5.00 10.00 15.00 20.00 25.00 .00 5.00 10.00 15.00 20.00 25.00 Time (months) Time (months) Group Group MCTC (+) CTC (+) CTC (+) censored MCTC (+) censored MCTC (-) CTC (-) CTC (-) censored MCTC (-) censored (a) (b) 1.00 1.00 0.80 0.80 0.60 0.60 0.40 0.40 0.20 0.20 0.00 0.00 .00 5.00 10.00 15.00 20.00 25.00 5.00 10.00 15.00 20.00 25.00 Time (months) Time (months) Group Group First course of First course of radiotherapy TNM stage I-II TNM stage I-II censored radiotherapy (normal) (normal) censored TNM stage III TNM stage III censored First course of radiotherapy First course of radiotherapy (adjust the strong) (adjust the strong) censored (c) (d) Figure 2: PFS analysis. of great signi“cance for NPC patients regardless of before with TNM staging. „erefore, expanding the sample size, and after operation. However, there is some controversy revalidating the correlation of CTC status with TNM is about the correlation between CTC and TNM staging. „e warranted in future experiments. studies of Li et al. [12, 13] clearly pointed out that the positive CTCs are also related to the prognosis of tumor patients. rate of CTC in NPC patients has no signi“cant correlation Studies have reported that CTC-positive breast cancer, Cumulative survival rate (%) Cumulative survival rate (%) Cumulative survival rate (%) Cumulative survival rate (%) Journal of Oncology 7 Table 5: COX univariate analysis. 95.00% CI Items B SE Wald Df P Exp (B) Upper Lower CTC 1.091 0.5782 3.566 1 0.0594 2.978 0.9608 9.241 MCTC 1.126 0.5363 4.408 1 0.0361 3.083 1.078 8.818 Age −0.969 0.5726 2.871 1 0.0921 0.3791 0.1245 1.164 Gender −0.042 0.5094 0.0071 1 0.9342 0.9597 0.3538 2.601 TNM stage 1.418 0.6383 4.947 1 0.0263 4.130 1.183 14.41 T 0.4501 0.4942 0.8293 1 0.3622 1.568 0.5952 4.130 N 0.6042 0.4871 1.538 1 0.2156 1.829 0.7046 4.746 Tumor state 0.4646 0.4862 0.9120 1 0.3400 1.591 0.6131 4.127 First course of radiotherapy 1.411 0.5082 7.733 1 0.0062 4.099 1.513 11.10 Table 6: COX multifactor analysis. 95.00% CI Subject B SE Wald Df P Exp (B) Upper Lower MCTC 1.673 0.7799 4.600 1 0.0319 5.327 1.155 24.57 TNM stage 1.094 0.4583 5.696 1 0.0169 2.986 1.216 7.332 First course of radiotherapy 0.675 0.5185 1.693 1 0.1932 1.963 0.7110 5.425 colorectal cancer, and liver cancer have a poor prognosis and there are many important tissues and organs around the shorter survival time [14–17]. In nasopharyngeal carcinoma, nasopharynx. Intensive radiotherapy will aggravate the Zhang [18] found that after 2–4 cycles of chemotherapy, the damage to the surrounding tissues and will also have a certain impact on the prognosis. Another reason is the CTC count of NPC was reduced remarkably, and the change in CTC number was correlated with treatment response. small number of cases in this study, the short follow-up time, and the lack of analysis of the long-term adverse reactions of Qian [19] reported that the reduction of CTC was related to the treatment effect of patients with advanced NPC, and the the patients. /erefore, whether the first radiotherapy positive CTC before treatment was an independent risk method affects the prognosis of NPC recurrence needs to be factor for poor prognosis. Yu [20] reported that CTC was further explored. /e COX analysis screened out that MCTC associated with poor survival in NPC patients, which may be and TNM staging were independent risk factors affecting the an independent prognostic factor affecting NPC. In our prognosis of local recurrence or local residual NPC after study, the positive rates of CTC and MCTC detected in endoscopic resection, suggesting that MCTC is an important patients with nasopharyngeal carcinoma after nasal en- factor affecting the prognosis of NPC patients. doscopy were significantly lower than those before treat- In conclusion, the detection of CTC is helpful for the prognosis evaluation after endoscopic nasal resection of ment, suggesting that CTC and MCTC counts changed before and after endoscopic nasal resection. For transnasal local recurrence or local residual NPC. MCTC is an im- endoscopic resection, CTC and MCTC are also potential portant factor affecting the prognosis of patients with NPC. indicators to monitor the prognosis of NPC patients. /e PFS of patients with different clinical characteristics was Data Availability analyzed, and the results showed that the PFS of CTC- positive NPC patients was significantly shorter than that /e data used to support the findings of this study are of CTC-negative patients, and the PFS of MCTC-positive available from the corresponding author upon request. NPC patients was significantly shorter than that of MCTC- negative patients. /e PFS of NPC patients in TNM stage (I- Conflicts of Interest II) was significantly longer than that of TNM stage (III) patients, and the PFS of NPC patients whose first radio- /e authors declare that they have no conflicts of interest. therapy mode was conventional was significantly longer than that of patients whose first radiotherapy mode was Acknowledgments enhanced. 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Published: Sep 12, 2022

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