上皮间质转化相关表型的循环肿瘤细胞在胰腺癌中的检测及相关性研究

发布时间：2018-11-21 11:26

【摘要】：[目的]1.研究循环肿瘤细胞(Circulating tumor cell,CTCs)在健康人和胰腺癌患者中的差异。2.评估CTCs与胰腺癌临床病理和预后相关性。3.探讨上皮-间质转化(Epithelial-Mesenchymal Transition,EMT)相关表型的CTCs随治疗过程的动态变化。4.通过比较胰腺癌原位肿瘤、外周血CTCs和转移淋巴结中上皮标志物和间质标志物的表达情况对胰腺癌EMT进行研究。[方法]1.采用芯片法对胰腺癌患者外周血CTCs进行上皮标志物E-cadherin和间质标志物Vimentin的染色,分析不同表型下CTCs与临床病理预后的相关性,并探讨手术、化疗过程中不同表型CTCs的动态变化。2.应用免疫组化法对确诊胰腺导管腺癌患者的原位肿瘤及转移淋巴结中上皮标志物E-cadherin和间质标志物Vimentin进行染色及评分,分析EMT与临床病理及预后的相关性;并结合芯片法中CTCs的上皮间质标志物表达情况对胰腺癌侵袭转移过程中EMT的变化进行研究。[结果]1.第一部分研究共纳入56名研究对象,包括46名胰腺导管腺癌患者和10名健康人,研究发现健康人和胰腺癌患者外周血CTCs数目的临界值为8.5/2ml,未转移胰腺癌和转胰腺癌患者外周血CTCs数目的临界值为49.5/2ml;CTCs总数和M表型CTCs与胰腺癌的分化程度、脉管瘤栓和病理分期具有显著的相关性;当CTCs数目超过47/2ml(总数)、11/2ml(E表型)、17/2ml(E+M表型)和35/2ml(M表型)时,提示患者预后不良。CTCs的数目与手术没有相关性,化疗会导致CTCs计数的显著下降,但是在化疗2个周期后,CTCs计数无明显改变。2.第二部分研究共纳入35名研究对象,通过免疫组化分析发现,相较于癌旁组织,胰腺癌原位组织中E-cadherin表达下降,Vimentin表达升高,二者相比具有统计学意义(P=0.000和P=0.003)。胰腺癌原位组织中E-cadherin和Vimentin的表达情况与临床病理预后无相关性。胰腺癌原位组织中E-cadherin高表达比例为68.6%,Vimentin高表达比例为37.1%,而CTCs中表达Vimentin的比例为78.6%。间质表型的CTCs随着化疗进行而出现比例的升高。[结论]1.CTCs与胰腺癌的分化程度、脉管瘤栓和病理分期具有相关性,可以反映病情的严重程度,并与患者的预后具有密切关联。2.胰腺癌在转移过程中上皮表型和间质表型呈现动态变化,循环肿瘤细胞的上皮表型和间质表型随着化疗呈现动态变化。
[Abstract]:[objective] 1. To study the difference of circulating tumor cell (Circulating tumor cell,CTCs between healthy and pancreatic cancer patients. 2. 2. To evaluate the correlation between CTCs and clinicopathology and prognosis of pancreatic cancer. To investigate the dynamic changes of CTCs associated with epithelial-mesenchymal transformation (Epithelial-Mesenchymal Transition,EMT) with the course of treatment. 4. The expression of epithelial markers and interstitial markers in pancreatic carcinoma in situ tumor, peripheral blood CTCs and metastatic lymph nodes were compared to study the expression of EMT in pancreatic carcinoma. [methods] 1. The epithelial marker E-cadherin and interstitial marker Vimentin were stained with microarray in peripheral blood of patients with pancreatic cancer. The correlation between CTCs and clinicopathological prognosis was analyzed under different phenotypes. Dynamic changes of CTCs in different phenotypes during chemotherapy. 2. The epithelial marker E-cadherin and interstitial marker Vimentin in situ tumor and metastatic lymph nodes of patients with pancreatic ductal adenocarcinoma were stained and scored by immunohistochemical method. The correlation between EMT and clinicopathology and prognosis was analyzed. The changes of EMT during invasion and metastasis of pancreatic carcinoma were studied by using the expression of epithelial interstitial markers of CTCs in microarray method. [result] 1. The first part of the study included 56 subjects, including 46 patients with pancreatic ductal adenocarcinoma and 10 healthy people. The critical value of peripheral blood CTCs in healthy and pancreatic cancer patients was 8.5 / 2ml. The critical value of peripheral blood CTCs was 49.5 / 2 ml in patients with non-metastatic and metastatic pancreatic cancer. The total number of CTCs and M-phenotypic CTCs were significantly correlated with the degree of differentiation, vascular embolus and pathological stage of pancreatic cancer. When the number of CTCs exceeded that of 47/2ml (total), 11/2ml (E phenotype), 17/2ml (E M phenotype (M phenotype) and 35/2ml (M phenotype), it suggested that the prognosis of patients was poor. There was no correlation between the number of CTCs and surgery. Chemotherapy resulted in a significant decrease in CTCs count, but no significant change in CTCs count after 2 cycles of chemotherapy. In the second part of the study, 35 subjects were studied. Immunohistochemical analysis showed that the expression of E-cadherin decreased and the expression of Vimentin increased in the in situ tissues of pancreatic cancer compared with the adjacent tissues. There was significant difference between the two groups (P 0. 000 and P 0. 003). There was no correlation between the expression of E-cadherin and Vimentin and the clinicopathologic prognosis of pancreatic carcinoma in situ. The proportion of high expression of E-cadherin in pancreatic carcinoma was 68.6% and that of Vimentin was 37.1%, while the proportion of Vimentin expression in CTCs was 78.6%. The proportion of interstitial phenotypic CTCs increased with chemotherapy. [conclusion] 1.CTCs is correlated with the degree of differentiation, vascular embolus and pathological stage of pancreatic cancer, which can reflect the severity of the disease and is closely related to the prognosis of the patients. 2. The epithelial phenotype and interstitial phenotype of pancreatic carcinoma showed dynamic changes during metastasis, and the epithelial phenotype and interstitial phenotype of circulating tumor cells showed dynamic changes with chemotherapy.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.9

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