PTEN基因转染联合奥沙利对胆管癌细胞生长的影响

Inhibitory effects of PTEN gene transfection combined with L-OHP on proliferation of human cholangiocarcinoma cells

目的：观察脂质体介导PTEN基因转染人胆管癌细胞（QBC939），联合化疗药物奥沙利铂（L—OHP）对胆管癌细胞生长的影响，探索人类胆管癌的生物治疗方法。方法：将携带PTEN基因的真核表达载体pBP-PTEN和不含该基因的空载体转染胆管癌QBC939细胞，嘌呤霉素抗性筛选克隆、扩增培养，S-P免疫组化法检测转染前后PTEN阳性表达率。实验分组为QBC939、QBC＋L-OHP、PTEN-QBC和PTEN＋L-OHP 4组，以MTT法检测癌细胞生长活性，透射电镜扫描观察转染前后及联合奥沙利铂后细胞的超微结构变化，流式细胞仪分析转染前后细胞周期变化和凋亡情况，体外细胞侵袭力抑制试验观察转染及用药前后细胞侵袭力的变化。结果：PTEN基因转染后QBC939细胞稳定表达、PTEN阳性表达率升高（P＜0．05）；基因转染后肿瘤细胞活性下降（P＜0．05）；细胞周期G1～S期抑制、细胞凋亡率增加（P＜0．01）；透射电镜下显示细胞较成熟、分化好、线粒体增多；细胞侵袭力明显抑制（P＜0．05），其中以PTEN＋L-OHP抑制作用最强，L-OHP抑制作用次之。结论：PTEN基因转染生物治疗联合奥沙利铂化疗对人胆管癌细胞生长具有显著的抑制作用。

Objective： To investigate the inhibitory effects of PTEN gene transfection combined with L-OHP on human cholangiocarcinoma cell line, QBC939, providing a new method for gene therapy of human biliary duct carcinoma. Methods： A eukaryotic expression vector containing PTEN gene was transfected into human QBC939 cells under mediation of lipofectamine and positive cell clones were selected and amplified. Expression of PTEN gene was detected by immunohistochemistry. MTT test was used to determine the in vitro activity of cells, electron microscope was applied to observe cell uhrastructure, and flow cytometry was used for determining the cell cycle and apoptosis. In vitro test was used to study the invasive ability of cells before and after treatment. Results： After transfected with PTEN gene, QBC939 cells had a higher expression of PTEN gene （ P 〈 0.05 ）, a decreased activity （ P 〈 0.05 ）, an arrest of G1 -S phase, and an increased apoptotic rate （P 〈0.01 ）. Electron microscope showed that maturity of cells and increased well-differentiated mitochondria. The aggressiveness of the cells was obviously inhibited （P 〈 0.05 ）. Conclusion ： PTEN gene transfection combined with L-OHP has obvious inhibitory effect on cholangiocarcinoma cell line in vitro.