Abstract
目的 探讨脂质体介导PTEN基因转染人胆管癌细胞(QBC939),联合化疗药物奥沙利铂(L-OHP),分别进行人胆管癌细胞体外培养和体内接种生长,观察分析该基因对胆管癌细胞生长的抑制情况,探索人类胆管癌的生物治疗的可行性和方法.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.转胆管癌QBC939细胞,嘌呤霉素抗性筛选克隆、扩增培养.绘制细胞生长曲线及MTT细胞活性观察;利用免疫组化检测转染前后PTEN阳性表达率;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;流式细胞分析细胞的周期变化和细胞凋亡情况;体外细胞侵袭力抑制试验;裸鼠体内肿瘤生长,病理学及电镜扫描的观测.结果PTEN基因转染后QBC939细胞稳定表达,阳性表达率升高(P＜0.05);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);透射电镜细胞较成熟、分化好;细胞侵袭力明显抑制(P＜0.05);裸鼠体内肿瘤未转染组生长早,加入奥沙利铂后生长受抑制(P＜0.05),病理证实为腺癌.结论1.脂质体成功将PTEN基因转染人胆管癌QBC939细胞,并稳定表达.2.PTEN基因转染后细胞生长速度无明显变化;MTT实验活性细胞数有所下降.3.转PTEN基因后的胆管癌细胞超微结构变化显示线粒体增多,细胞较成熟、分化好;流式细胞议分析,细胞周期G1期被抑制,细胞凋亡多,侵袭力受到抑制;4.接种转PTEN基因裸鼠的体内成瘤显著抑制;6.转基因的生物治疗联合奥沙利铂(L-OHP)对人胆管癌细胞生长具有显著的抑制作用。
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