阻断Sonic Hedgehog信号对不同人肝癌细胞生长的影响  

Effect of Sonic Hedgehog signaling blockade on growth of hepatocarcinoma cells

作　　者：刘爱梅[1] 余功旺 黄莉霞[1] 孙艳[1] 迟作华[2] 

机构地区：[1]广东药学院生命科学与生物制药学院,广东广州510006 [2]广东药学院临床医学院,广东广州510006

出　　处：《中国病理生理杂志》2016年第2期208-214,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81000923);广东省科技计划(No.2013B021800089)

摘　　要：目的:研究阻断Sonic Hedgehog(Shh)信号对不同人肝癌细胞生长的影响,探讨阻断Shh信号抑制肝癌细胞生长的机制。方法:RT-PCR法检测Shh信号分子在3株人肝癌细胞(BEL-7402、Huh7和HepG2)中的表达,并检测Shh阻断抗体作用后BEL-7402细胞Shh信号效应分子表达变化;MTT法检测人肝癌细胞增殖活性;流式细胞术检测人肝癌细胞凋亡;Western blot检测凋亡相关蛋白表达。结果:Shh信号分子在3株人肝癌细胞中均有表达,Shh阻断抗体可以下调Shh信号效应分子patched(Ptch)、Gli1和Gli2的表达;Shh阻断抗体可以抑制3株肝癌细胞生长,增加G0/G1期细胞,并诱导细胞凋亡;Shh阻断抗体作用后,BEL-7402细胞pro-caspase-3、pro-caspase-8和pro-caspase-9蛋白表达水平下降,cleaved caspase-3、cleaved caspase-8和cleaved caspase-9蛋白表达水平升高。结论:阻断Shh信号可抑制Shh高表达的人肝癌细胞生长,阻滞细胞周期于G0/G1期,并诱导肝癌细胞凋亡。AIM ： To investigate the effect of Sonic Hedgehog （Shh） signaling blockade on the growth of hema- tocarcinoma cells and underlying mechanisms. METHODS： The expression of Shh signaling molecules in hematocarci- noma ceil lines BEL-7402, Huh？ and I4epG2 was detected by RT-PCR. The cell viability was detected by MTY assay. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of apoptosis-related proteins was determined by Western blot. RESULTS： Shh signaling molecules were all expressed in BEL-7402, Huh7 and HepG2 cells. The mRNA expression of Patched （ Ptch）, Glil and Gli2 was down-regulated by anti-Shh antibody. Blockade of Shh signaling pathway inhibited the proliferation of hepatocarcinoma cells with increasing cells in G0/G1 phase and induced the apoptosis of hepa- tocarcinoma ceils. Treatment with anti-Shh antibody down-regulated the protein expression of pro-caspase-3, pro-caspase-8 and pro-caspase-9, while up-regulated the protein levels of cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9 in BEL-7402 cells. CONCLUSION： Blockade of Shh signaling pathway inhibits the growth of hepatocarcinoma at different levels by cell cycle arrest and inducing apoptosis of hematocarcinoma cells.

关 键 词：肝癌细胞 Sonic HEDGEHOG 细胞凋亡 

分 类 号：R730.23[医药卫生—肿瘤]