中文摘要：

目的探讨微RNA－26a（miR26a）在肝内胆管癌中的表达情况，分析其作用机制及临床意义。方法收集46例肝内胆管细胞癌及癌旁组织标本，荧光定量逆转录－PCR法检测miR26a的表达，分析其与临床病理特征及预后的关系。肝内胆管癌细胞株HCCC－9810及RBE分别用lipo—fectamine2000转染miR26a仿品及miR26a抑制剂。CCK－8法测生长曲线，划痕及transwell实验分析迁移及侵袭能力，流式细胞仪测细胞周期，蛋白印迹检测分析其可能机制。结果miR－26a在肝内胆管癌组织中的表达较癌旁组织上调（P〈o．05h且有血管侵犯、TNMⅢ～Ⅳ期和有淋巴结转移组上调更为明显（P〈O．05）。miR－26a的变化与性别、年龄、肿瘤数目、肿瘤直径、肿瘤包膜、肿瘤分化的相关性差异无统计学意义（P〉0．05）。肝内胆管癌细胞HCCC-9810的miR-26a表达水平上调可以促进增殖及侵袭，加快G0／G1期到s期转换，抑制PTEN进而激活AKT信号通路。肝内胆管癌细胞RBE的miR-26a表达水平下调可以抑制增殖及侵袭，导致G0／G1期阻滞，PTEN表达增加而AKT激活受抑，PTENmRNA表达水平与miR－26a表达水平负相关（r＝-0．8272，P〈0．01）。miR26a高表达组总体生存率较低表达组差（P〈0．05）。miR26a高表达、多发肿瘤及有淋巴结转移是影响肝内胆管癌患者总体生存的独立风险因素（P〈0．01）。结论miR-26a在肝内胆管癌组织中高表达，可以影响肝内胆管癌的临床病理特征及总体生存率，并可能通过PTEN／AKT通路促进肿瘤的增殖及转移。

英文摘要：

Objective To study the expression and significance of miR 26a in intrahepatic cholangiocarcinoma. Methods The expression of miR-26a in 46 intrahepatic cholangiocarcinoma（ICC） tissues and peritumoral tissues was detected by quantitative real time polymerase chain reaction （qRT- PCR）. The intrahepatie cholangiocarcinoma cell line HCCC-9810 and RBE were transfected with miR- 26a mimics and miR 26a inhibitors, respectively, by lipofectamine 2000. The growth curves were con- structed by the CCK-8 method. The migration and invasion ability was demonstrated by wound healing and transwell assay. The cell cycle was analyzed by flow cytometry. The potential mechanism was il- lustrated by Western blotting. Results For the 46 ICC tissues and peritumoral tissues,miR 26a levels were significantly higher in the tumor tissues than in the peritumoral tissues （P〈0.05）. Vascular in vasion, TNM III~ IV stage and lymph node metastasis were significantly associated with high miR 26a expression levels （P〈0.05）, but gender, age, tumor amounts, tumor encapsulation, tumor di- ameter and tumor differentiation showed no significant association （P〉0.05）. Enhanced cell prolifer ation, migration and invasion ability, accelerated G0/G1 phase to S phase transition, activated AKT by PTEN suppression were observed in HCCC-9810 cells with up regulation of miR 26a. Conversely, cell proliferation, migration and invasion ability was inhibited, G0/G1 phase was blocked and AKT was restrained by PTEN increase with down regulation of miR-26a in RBE cells. PTEN mRNA in- versely correlated with the miR-26a level （r=-0. 8272, P〈0.01）. Patients with a high miR-26a expression had significantly poorer overall survival （P〈0.05）. A high miR-26a expression, multiple tumors and lymph node metastasis were independent prognostic factors of overall survival （P〈0.01）. Conclusion Overexpression of miR 26a in intrahepatic cholangiocarcinoma correlated with clinicopath- ological features and overall survival, and it potentially