Detection And Clinical Significance Of Circulatingtumor Cells In Peripheral Blood Of The Patients With Esophageal Squamous Cell Carcinoma

Objective：Esophageal carcinoma (EC) is the common alimentary canalcancer with poor prognosis and low5-year survival rate. Most esophagealcancer patients die as a result of tumor metastasis to distant organs.Circulating tumor cells (CTC) of blood may be an important reason of its highrecurrence and metastasis rate. The CTC detection of the peripheral blood isimportant for the judgement of the recurrence and the direction of clinictherapy in esophageal cancer patients. Epithelial cell adhesion molecule（Ep-CAM），which is also know as human epithelial antigen （HEA），isbroadly expressed on cells of epithelial origin and on epithelial derived tumorcells，and it not anly play a very important role in cell signaling、cellcycleregulation、adhesion、proliferation and differentiation，and in the organs whenthey formation and ma intenance，but also have a variety of function with thetumor occurrence development，transfer.In this study，the expression of the tumor cells which is showed theEP-CAM positive in peripheral blood of the patients with esophagealsquamous cell carcinoma was detected by lymphocyte separation and flowcytometry (FCM)。We appraisal the tumor cells by immune cells chemicalmethod.The purpose of this study is to explore the different expression ofEP-CAM positive circulating tumor cells before and after surgery or Beforeand after chemotherapy in patients with esophageal patients，and to analysisthe relationship between the expression of EP-CAM positive circulating tumorcells in patients with esophageal patients and clinical pathology characteristic，and explore the clinical significance of its detection.Methods:30surgical patients with esophageal squamous cell carcinoma came from Hebei Province People’s Hospital and HeBei Medical University4th Hospitalat the time in Sep2010-May2011. The5ml blood was obtained beforesurgery a day and one hour and ten days after operation.The result of patientsconfirmed by pathology were squamous cell carcinoma；30non-operativepatients with esophageal squamous cell carcinoma came from Hebei ProvincePeople’s Hospital Thoracic Surgery Department and Medical OncologyDepartment at the time in Sep2010-May2011. The5ml blood was obtainedbefore chemotherapy a day and five days after chemotherapy. The result ofpatients confirmed by pathology were squamous cell carcinoma.10bloodcame from healthy volunteers as normal control group. The peripheral bloodmonoclonal cells were separated from blood of all subjects by lymphocyteseparation. The expression of CD45~－EP-CAM~＋circulating tumor cells weredetected by the flow cytometry (FCM)and were evaluated by immune cellschemical method.We measured the expression of CD45~－EP-CAM~＋circulating tumor cells in the peripheral blood of all experimental subjects. Weanalyse difference expression of CD45~－Ep-CAM~＋circulating tumor cells inthe peripheral blood before and after surgery or Before and afterchemotherapy， and discuss the relationship between the expression ofEP-CAM positive cycle tumor cells in patients with esophageal patients andclinical pathology characteristic.Statistical Methods: all data were to analyzed by SPSS13.0statisticalsoftware for Х~2test and t tests and analysis of variance and nonparametrictest,α=0.05was differences significant standards， P value <0.05wasstatistically significant.Results:1The expression of circulating tumor cells in the peripheral blood ofhealthy volunteers and the patients with esophageal squamous cell carcinoma：The expression of CD45~－EP-CAM~＋circulating tumor cells in peripheralblood of the patients with esophageal squamous cell carcinoma wereSuccessful detected by the flow cytometry (FCM)，but cell numbers were less.The cytoplasm of EP-CAM~＋circulating tumor cells was tan cytoplasm byimmune cells chemical method.We did not detect EP-CAM~＋circulating tumorcells in10cases healthy volunteers，and no EP-CAM~＋circulating tumor cellsin5cases of60Patients with esophageal squamous cell carcinoma beforesurgery or chemotherapy，The positive rate of EP-CAM~＋circulating tumorcells was significantly differences between the two groups (X~2=37.507，P=0.000).2The expression of circulating tumor cells in the patients with esophagealsquamous cell carcinoma before surgery and one hour after surgery and tendays after surgery：The expression of circulating tumor cells in three groupswas difference(F=21.573，P=0.000). The expression of EP-CAM~＋circulatingtumor cells in three groups were：12.067±6.943（before surgery） and12.100±6.630（one hour after surgery）and8.433±4.576（ten days aftersurgery），the expression of circulating tumor cells was no significant betweenbefore surgery group and one hour after surgery group（P=0.983）,and wassignificant between before surgery group and ten days after surgery group（P=0.024），and was significant between one hour after surgery group and tendays after surgery group（P=0.023).3The expression of circulating tumor cells in the peripheral blood of30non-operative patients with esophageal squamous cell carcinoma before andafter chemotherapy：The expression of Ep-CAM~＋circulating tumor cells in theperipheral blood before chemotherapy was significantly higher than afterchemotherapy.the two groups expression of EP-CAM~＋circulating tumor cellsin the peripheral blood were：18.300±3.385（before chemotherapy）and11.067±2.518（after chemotherapy），there is significant differences at theexpression level,(t=19.366，P=0.000).4The relationship between the expression of circulating tumor cells inpatients with esophageal patients before surgery and clinical pathologycharacteristic：Divided the patients into different groups according to thepathological features， there was no significant correlation between theexpression level of EP-CAM positive circulating tumor cells and the degree of differentiation (P>0.05); but there were significantly correlated with tumorsize, invasion depth of tumor and the tumor lymph node metastasis (P <0.001).5The relationship between the expression of circulating tumor cells inpatients with esophageal patients before surgery and tumor stage：Divided thepatients into different groups according to the tumor stage，The expression ofEp-CAM＋circulating tumor cells was significantly between two groups：7.938±6.126（stageⅡ）and16.786±4.388（stageⅢ），there is significantdifferences at the expression level,(Z=-3.655，P=0.000).Conclusions:1This study can find circulating tumor cells in peripheral blood of patientswith esophageal squamous cell carcinoma by the detection of EP-CAM.2The number of circulating tumor cells in peripheral blood of patientswith esophageal squamous cell carcinoma was no difference before and aftersurgery at short time，but the number of circulating tumor cells decreased atlong time.3Chemotherapy can reduce the number of circulating tumor cells inblood.4There is significant correlation between the number of circulating tumorcells and tumor stage，tumor stage the later, the number of circulating tumorcells the more.