特异性阻断Shh信号通路对肝细胞癌细胞系Hep-3B增殖和凋亡的影响

目的 观察阻抑胚胎发育相关信号通路Sonic hedgehog(Shh)对人肝癌细胞系Hep-3B增殖和凋亡的影响.方法 将实验细胞分为转染组、隐性对照组和空白对照组.设计针对Shh信号通路中Smo基因的siRNA,通过Lipedectemin2000将其转染进Hep-3B细胞株,噻唑蓝(Mar)比色法检测沉默Smo基因后第0、8、16、24和48 h的细胞数和MTr的D(入)值;流式细胞术检测沉默Smo基因48 h细胞凋亡.结果 转染组与空白对照组比较,加入siRNA后,8 h时细胞增殖数无明显减少,16、24和48 h时细胞数明显减少(P<0.01),24和48 h时细胞存活率差异有统计学意义.阴性对照组与空白对照组比较,差异无统计学意义(P>0.05);Hep-3B细胞经siRNA作用48 h后,可见亚二倍体核型峰-凋亡峰.其中,转染组凋亡率为30%,阴性对照组与空白对照组未见凋亡峰.结论 Shh信号分子对肝细胞癌的增殖起维持作用;通过特异性阻断该信号通路,可以抑制肝癌细胞增殖,并促进细胞凋亡.

Effect of sonic hedgehog signal pathway blocking on the proliferation and apoptosis of hepatocellular carcinoma cell line Hep-3B

Objective To investigate the effect and mechanism of eyclopamine, the inhibitor of sonic hedgehog (Shh) signaling pathway, on the proliferation and apoptosis of hepatocellular carcinoma cell line Hep-3B. Methods The Hep-3B cells were cultured in vitro,and the MTT assay was used to ex-amine the antiprolfferative effect of siRNA. Flow cytometry (FCM) was used to examine the effects of ap-optosis of Hep-3B cells. Results Compared with the negative and blank control groups, the transfection group showed marked decrease of proliferation at 16,24 and 48 h and about 30% Hep-3B cells were apop-totic at 48 h after Smo was silenced by siRNA. Conclusion Shh signaling pathway is essential for the maintaining of hepatocellular carcinoma cell proliferation. Blocking shh signaling can inhibit the growth of hepatoceUular carcinoma cells and induce apoptosis.