阿霉素诱导胰腺癌细胞上皮-间质转化的研究-复旦大学.docVIP

阿霉素诱导胰腺癌细胞上皮-间质转化的研究 复旦大学上海医学院05级临床医学八年制华山班 刘恋 指导教师 复旦大学上海医学院病理学系 刘秀萍 【摘要】 目的 在临床上，化疗药物在除了治疗作用外，还可引起肿瘤恶性程度增高。最近，上皮间质转化（epithelial-mesenchymal transition, EMT）在肿瘤进展过程中的作用引起了人们的注意。我们关心的是短时间应用阿霉素（Adriamycin, ADM）是否会引起肿瘤细胞发生EMT，明确Twist1是否为调控阿霉素诱导胰腺癌细胞EMT的关键转录因子。 方法 采用倒置相差显微镜和Western blotting法分别从细胞形态和标志性蛋白分子（E-cadherin、CK19、α－SMA和β-vimentin）的表达情况，观察加入阿霉素24、36、48和72 h后，胰腺癌细胞株（BxPC-3）发生是否发生EMT及其时间点。采用Western Blot检测阿霉素诱导BxPC-3发生EMT过程中Twist1的表达情况。 结果 治疗剂量阿霉素处理48小时后，BxPC-3细胞形态发生EMT的改变。上皮细胞标志性蛋白分子E-cadherin和CK19表达减少，间质细胞标志性蛋白分子α－SMA和β-vimentin表达增加。Twist1在阿霉素诱导BxPC-3发生EMT变化过程中表达不增高。 结论 治疗剂量阿霉素早期即能诱导BxPC-3发生EMT，Twist1不是关键转录因子。 【关键词】 EMT；胰腺癌； ADM Adriamycin-Induced Epithelial-Mesenchymal Transition in Pancreatic Cancer Cells 【Abstract】 Purpose: Besides its therapeutic effects, chemotherapeutic agents also enhance the malignancy of treated cancers in clinical situations. Recently, epithelial-mesenchymal transition (EMT) has attracted attention in studies of tumor progression. We aimed to test whether transient Adriamycin treatment induces EMT in pancreatic cancer cells, and elucidate the role of Twist1, the master regulator of EMT, in this process. Experimental Design: In BxPC-3 cells, EMT after Adriamycin treatment was examined in vitro by inverted phase contrast microscope and Western blotting at 24, 36, 48, 72hours, to examine morphologic and molecular changes respectively. The Adriamycin-induced Twist1 expression was examined by Western blotting. Results: We showed in vitro that Adriamycin induced EMT at 48h in morphology and 36h in molecular. The expression of Twist1 did not increase in the process of ADM induced EMT. Conclusions: ADM induces EMT of pancreatic cancer cells very early. Twist1 is not the key regulator of ADM induced EMT of pancreatic cancer cells. 【Key Words】 EMT; Pancreatic cancer; ADM 上皮－间质转化（Epithelial-Mesenchymal　Transition，EMT)，胚胎发育中形态生发的一个关键过程，是具有极性的上皮细胞转换成具有活动能力、能够在细胞基质间自由移动的间质细胞的过程。它以上皮细胞极性的丧失及间质特性的获得为重要特征,具体包括: 细胞黏附分子( E－钙黏蛋白)表达减少；角