干扰RN181基因的表达抑制SMMC7721肝癌细胞的凋亡

Interferencing RN181 gene inhibits the apoptosis of hepatocellular carcinoma SMMC7721 cells

[目的]探讨稳定干扰RN181基因对顺铂诱导的SMMC7721肝癌细胞凋亡的影响。[方法]采用RN181干扰慢病毒感染SMMC7721,流式分选出带绿色荧光的细胞,Western Blot鉴定RN181干扰情况;采用DAPI染色法、Western Blot检测RN181对顺铂诱导SMMC7721肝癌细胞凋亡的变化情况。[结果]重组细胞株荧光表达良好,7721KD中RN181蛋白表达量明显低于其对照组。DAPI染色法观察到在顺铂作用下,7721KD的细胞凋亡阳性率低于其对照组(P<0.05)。WB检测到7721KD activated caspase-3、cleaved PARP的表达量低于对照组,而pro-caspase-6、pro-caspase-8的表达量与对照组相比升高。[结论]稳定干扰RN181的表达能够抑制顺铂诱导的SMMC7721细胞凋亡,其机制可能是通过参与死亡受体凋亡途径。

[ Objective]To study the effects of RN181 gene on the apoptosis of the SMMC7721 cell line induced by cisplatin. [ Methods] The SMMC7721 cells infected by the RN181 knocking- down letivirus were sorted through FACS and identified by Western Blot . The effects of RN181 on cisplatin - induced apoptosis were detected by DAPI staining and Western Blot. [ Results] We verified protein expression of RN181 in the recombinant cell lines by microscope and WB ; Under the treatment of cisplatin, the ratio of cell apoptosis was decreased in recombinant cell line 7721KD （ P 〈 0. 05 ） ; Compared with the control, the expression of activated easpase - 3 and cleaved PARP were lower but pro - caspase - 5 and pro - caspase - 8 were higher in 7721KD. [ Conclusion] Interferencing RN181 expression can inhibit the cisplatin- induced apoptosis of SMMC7721 cell line. Its mechanism may be through participating in death receptor apoptosis pathway.