RN181过表达影响SMMC7721肝癌细胞凋亡的初步研究

目的构建过表达RN181基因的SMMC7721重组细胞株,研究RN181对顺铂诱导肝癌细胞凋亡的影响,并初步探讨其凋亡机制。方法构建高表达RN181逆转录病毒感染SMMC7721细胞,显微镜观察其细胞株荧光强弱,Western blot鉴定RN181蛋白表达水平;采用DAPI染色法、Western blot检测RN181对顺铂诱导SMMC7721细胞株凋亡的变化情况。结果重组细胞株荧光表达良好,7721RN181细胞中RN181蛋白表达量高于其对照,差异有统计学意义(P0.05)。DAPI染色法观察到在顺铂作用下,7721RN181的细胞凋亡阳性率高于其对照,差异有统计学意义(P0.05);Western blot检测到7721RN181的activated caspase-3、cleaved PARP的表达量高于对照组,procaspase-6,pro-caspase-8的表达量低于对照组,差异均有统计学意义(P0.05)。结论 RN181表达能促进肝癌细胞的凋亡,其可能通过参与死亡受体凋亡途径来促进肝癌细胞凋亡。进而推测RN181基因可能成为肝癌靶向治疗的一个新靶点。

Overexpression of RN181 accelerate cisplatin-induced apoptosis of hepatocellular carcinoma SMMC7721 cells

Objective To establish recombinant cell line with overexpression of RN181 and investigate the effect of RN181 on cisplatin-induced apoptosis of SMMC7721 cells. Methods The SMMC7721 cells, transfected with RN181 over-expressing retrovirus, were identified through their fluorescence and protein levels of RN181. The effects of RN181 on cisplatin-induced apoptosis were evaluated by DAPI staining and Western blot. Results We verified protein expression of RN181 in the recombinant cell lines by microscope and Western blot （P〈0.05）. Under the treatment of cisplatin, cell apoptosis ratio was increased in recombinant cell line 7721RN181 （P〈0.05）. The expression of activated caspase-3, cleaved PARP were higher, pro-caspase-6 and pro-caspase-8 were lower in 7721RN181, compared with control （P〈0.05）. Conclusions RN181 could accelerate cisplatin-induced apoptosis of SMMC-7721 cells through death receptor apoptosis pathway. It could be regarded as a novel target for clinical diagnosis and gene therapy for HCC.