PARP-1抑制剂对人肝癌细胞HepG2增殖和迁移的抑制作用

Effects of Different PARP-Inhibitors on the Proliferation and Migration of Human Hepatocellular Carcinoma Cell Line HepG

目的 研究证实聚腺苷二磷酸核糖聚合酶-1（poly ADP ribose polymerase,PARP-1）抑制剂对肿瘤细胞的增殖、凋亡和侵袭有影响,但对肝癌细胞生物学特性的影响尚不清楚,此研究的目的是观察3种不同的PARP-1抑制剂对人肝癌细胞株HepG2增殖、凋亡及迁移的影响及可能机制.方法 MTT检测体外不同浓度的PARP-1抑制剂AG014699,BSI-201,AZD-2281处理后HepG2细胞的增殖;随后选择抑制效果明显的抑制剂AG014699和BSI-201处理HepG2;Western印迹法检测HepG2细胞Casepase3、Casepase8、Bax、Bcl-2、PTEN、Timp3、MMP3蛋白的表达水平.Transwell实验检测AG014699和BSI-201对HepG2细胞迁移的影响.结果 AG014699,BSI-201,AZD-2281均具有抑制HepG2细胞增殖的作用,具有时间和浓度依赖性,作用HepG2细胞48 h后的Caspase3、Caspase8、Bax、PTEN、Timp3蛋白的表达水平随药物浓度的增加而增高,而Bcl-2和MMP3蛋白水平随药物浓度的增加而降低且与对照组相比有显著性差异（P〈0.01）.结论 在体外PARP-1抑制剂AG014699、BSI-201、AZD-2281明显抑制HepG2细胞的增殖,但AG014699和BSI-201展示较好的敏感性,同时二者诱导肝癌细胞凋亡和抑制肝癌细胞的迁移,这其中的机制可能与影响凋亡信号的通路以及迁移相关蛋白的表达有关.

Objective It has been confirmed that the inhibitors of poly ADP-ribose polymerase-1 （PARP-1） affect the proliferation,apoptosis and invasion of tumor cells.However,the impact of the inhibitors of PARP-1 on hepatoma cells remains unclear.The purpose of this study was to investigate the effect of three kinds of PARP-1 inhibitors on the proliferation,apoptosis and migration of hepatocellular carcinoma cells in vitro.Methods MTT assay was performed to detect the proliferation of HepG2 cells after treatment with PARP-1 inhibitors,AG014699,BSI-201,and AZD-2281.Thereafter,AG014699 and BSI-201,which had the most suppressive effect,were used to treat HepG2 cells.Western blot assay was used to detect the protein expression of asepase3,casepase8,Bax,Bcl-2,PTEN,tissue inhibitor of metalloproteinase 3 （TIMP3）,and matrix metalloprotease 3 （MMP3）.Transwell assay was performed to detect the effect of AG014699 and BSI-201 on the migration of HepG2 cells.Results AG014699,BSI-201 and AZD-2281 all could inhibit the proliferation of HepG2 cells in a time-and concentration-dependent manner.The protein expression levels of caspase3,caspase8,Bax,PTEN,TIMP3 increased and those of Bcl-2 and MMP3 decreased,with the increasing of the drug concentration,in HepG2 cells treated for 48 h,which showed a significant difference as compared with the control group （P〈0.01）.Conclusion All PARP-1 inhibitors AG014699,BSI-201,and AZD-2281 could significantly inhibit the proliferation of HepG2 cells,with AG014699 and BSI-201 having better sensitivity.AG014699 and BSI-201 could significantly induce the apoptosis and inhibit the migration of hepatoma cells,which may be related to the alteration of apoptosis-related signaling pathway and the expression of migration-related proteins.