A Study On Detection Of Circulating Tumor Cells In Esophageal Squamous Cell Carcinoma

ObjectiveThe research team has detected 61 cases of peripheral blood CTCs in patients with esophageal squamous cell carcinoma in 2014.Based on this study,the patients were first followed up to analyze the clinical guiding significance of CTCs on the prognosis of patients.Next,the expressions of CK8,CK18,CK19,EpCAM,Vimentin,and E-Cadherin in esophageal squamous cell carcinoma were detected.The expression differences of epithelial markers CK8,CK18,CK19,and EpCAM in esophageal squamous cell carcinoma and CTCs were examined to investigate esophageal scales.ETC in cancer CTCs.Finally,the typical CTCs images of patients with gastric cancer and esophageal squamous cell carcinoma were selected and their morphological(cytological)characteristics were analyzed.MethodsWe followed up the 61 ESCC patients who underwent CTCs testing in 2014 and used statistical methods to analyze the relationship between CTCs and patient outcomes.Immunohistochemistry was used to detect the expression of CK8,CK18,CK19 and EpCAM in 41 of all cases.The expression of CK8,CK18,CK19,and EpCAM in tumor tissues and CTCs was compared to discuss whether CTCs were suffered EMT.The morphological(cytological)characteristics of esophageal squamous cell carcinoma and gastric cancer CTCs isolated from the CTCBIOPSY?system were identified according to the morphological diagnostic criteria for CTCs prepared by our group.ResultsThe PFS in patients with CTCs-positive esophageal squamous cell carcinoma was significantly shorter than that in CTCs-negative patients,and PFS was negatively correlated with the number of CTCs.Multivariate analysis showed that CTCs count >2 was an independent prognostic indicator of tumor recurrence(hazard ratio[HR] 5.63,95% confidence interval [CI] 1.77-17.89;P=0.003).Subgroup analysis showed that in patients with II,I-IIIB,T3,and T3+T4,patients with CTCs >1 or >2had significantly shorter PFS than patients with CTCs≤1 or CTCs≤2;,patients with CTCs >0 had significantly shorter PFS than patients with CTCs ≤0 in phase III and T3+T4 patients.Immunohistochemistry results showed that EpCAM and CK8,CK18,and CK19 were co-expressed in primary tumor tissues and CTCs in 7 of 41 patients.The Celltracks? AutoPrep? system results showed that 7 patients did not express CK8,CK18,CK19,and/or EpCAM,but they were isolated by ISET technique and 3 of 7patients had CTCs in their peripheral blood.This indicateed that 3 patients had EMT-CTCs and were missed by Celltracks? AutoPrep? system.The expressions of CK8,CK18,CK19,EpCAM,Vimentin and E-cadherin were not significantly different in different CTCs count groups(P>0.05).The expression of CK8,CK18,and CK19 in esophageal squamous cell carcinoma was significantly correlated with the decrease of E-cadherin expression(P=0.01),and the decrease of E-cadherin expression was also significantly associated with the expression of Vimentin(P=0.03).The morphological(cytological)characteristics of typical gastric cancer and esophageal cancer CTCs images were initially described,and the accuracy of detected CTCs was verified by immunofluorescence.ConclusionThis is the first report that ISET detection of CTCs is an independent predictor of prognosis in patients with ESCC.CTCs testing may improve the accuracy of preoperative staging of ESCC.EMT in CTCs of ESCC patients may lead to low detection rate of Celltracks? AutoPrep? System CTCs.The morphological diagnostic criteria for CTCs proposed by this group are scientifically feasible;in addition to morphological studies,qualitative analysis of selected CTCs is needed to verify the nature of CTCs and provide a basis for further downstream research.