腺病毒介导的p27mt基因转染对肝癌细胞及裸鼠移植瘤的生长抑制作用

Inhibitory Effect of recombinant p27mt Adenovirus on Hepatic Carcinoma Cells and Transplanted Tumor in Nude Mice

目的:研究携带突变型p27的重组腺病毒(Ad-p27mt)基因转染对肝癌的体内、外抑制作用,并探讨其抑癌机制。方法:⑴将Ad-p27mt转染人肝癌细胞SMMC-7721,Western blotting检测P27、Bcl-2、Bax的表达;流式细胞仪(FCM)观察Ad-p27mt对肝癌细胞生长抑制作用。⑵Ad-p27mt直接注射入人肝癌裸鼠移植瘤中,绘制肿瘤生长曲线,计算肿瘤生长抑制率。结果:Ad-p27mt转染肝癌细胞后,细胞增殖明显受抑制,G0/G1期细胞比例增加,P27表达增强,Bax/Bcl-2比例增高,采用Ad-p27mt基因治疗肝癌裸鼠移植瘤生长受抑制,肿瘤生长抑制率达57.09%。结论:Ad-p27mt基因转染对肝癌具有较显著的体内、外抑制作用和诱导凋亡作用,其机制是通过增强P27表达,使细胞产生周期阻滞,诱导凋亡的机理可能是上调Bax/Bcl-2比例所引起。

Objective To investigate the inhibitory effect of Ad-p27mt on hepatic carcinoma in vitro and in vivo and its possible mechanism. Methods （1）Hepatic carcinoma cells SMMC-7721 were infected with Ad-p27mt, the expression of P27, Bcl-2, Bax were determined with Western blot. The Inhibitory effects of Ad-p27mt on the growth of hepatic carcinoma cells were evaluated with flow cytometry. （2）The therapeutic effects were evaluated with growth curves after injection of Ad-p27mt into transplanted tumor in nude mice ,and eaculated the inhibitory rates of tumor growth. Results After infected with Ad-p27mt, the proliferation of SMMC-7721 cells was markedly inhibited. The proportion of cells at G0/G1 phase was increased,the expression of P27 and Bax/Bel-2 protein ratio were increased significantly. The growth of tumors in gene therapy with p27mt were obviously inhibited, the inhibitory rate of tumor growth was up to 57.09%. Conclusion p27mt gene transfer mediated by adenovirus vector has significant inhibitory effect and inducing apoptosis on hepatic carcinoma in vivo and in vitro, its mechanism may be the upregulation of p27mt expression that cause the carcinoma cell cycle arrest, the apoptosis induction may be the upregulation of bax/Bel-2.