TPh对肝癌细胞HepG-2细胞周期、凋亡及相关因子表达的影响  

Effect of TPh on cell HepG-2 cell cycle,apoptosis and related gene expression of hepatocellular carcinoma

作　　者：菅若男 范蕾[1] 刘丽娟[1] 马琛 李凌[1] 

机构地区：[1]内蒙古医科大学药学院,内蒙古呼和浩特010110 [2]广州省珠海市高新区唐家湾镇卫生院,广东珠海519080

出　　处：《中国生化药物杂志》2017年第2期15-19,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：内蒙古自治区自然科学基金

摘　　要：目的研究新化学合成药物2-苯基-3,4-噻二唑[3,2-α]并嘧啶-7酮（TPh）对人肝癌细胞HepG-2增殖活性、细胞周期、凋亡及相关因子表达的影响,探讨其抗肿瘤作用及作用机制。方法采用MTT法检测不同浓度的TPh作用不同时间后对HepG-2细胞增殖抑制作用;以不同浓度的TPh处理HepG-2细胞一定时间后,采用Hoechst 33258染色法观察细胞凋亡形态;采用流式细胞仪检测TPh对人肝癌HepG-2细胞周期的影响;采用Western blot法检测HepG-2细胞Bcl-2、Bax蛋白表达情况。结果 MTT法实验结果表明：TPh对人肝癌HepG-2细胞增殖有抑制作用且呈剂量依赖性,随浓度增加抑制率逐渐加强,浓度为最小值12.5μg/mL,抑制率达50.9%（P〈0.01）。Hoechst 33258染色结果显示：在荧光显微镜下,正常的肝癌细胞呈浅蓝色荧光,凋亡的肝癌细胞呈亮蓝色荧光,且随药物浓度的增加,肝癌细胞凋亡增多。流式细胞仪检测TPh对人肝癌HepG-2细胞周期的结果显示：与空白对照组相比TPh各剂量组G0/G1期细胞比例逐渐增加,S期细胞比例有不同程度的减少,G2/M期细胞比例也有不同程度的减少;随着药物浓度的增加,凋亡的细胞数目也不断增多（P〈0.05）;Western blot结果显示：与空白组相比,TPh以药物浓度依赖性抑制Bcl-2蛋白的表达,并且Bax基因的表达增加,TPh各剂量组的Bcl-2/Bax的比值率明显下降,并呈现剂量依赖性（P〈0.05）。结论 TPh可以抑制HepG-2的增殖,促进其凋亡,诱导肝癌细胞HepG-2 G0/G1期阻滞,其作用机制可能与Bax蛋白表达增加、Bcl-2蛋白表达减少有关。Objective To investigate the effect of different concentrations of TPh on hepatoma cells,cell viability and and its possible mechanisms of anti-tumor.Methods The inhibitory rate of hepatoma cells and cell viability on different concentrations of TPh and time were measured by MTT assay;The morphological changes of apoptosis were observed by Hoechst 33258 straining;Cell cycle distribution was evaluated by flow cytometry（FCM）.The protein expression of Bcl-2 and Bax were detected by Western blot analysis.Results MTT assay showed that TPh inhibited the proliferation of HepG-2cells in a dose-dependent manner,and the inhibitory rate increased with the increase of concentration.The inhibitory rate was 50.9%（P〈0.01）.The results of Hoechst 33258 staining showed that the fluorescence intensity of hepatocellular carcinoma cells was light blue in fluorescence microscopy and bright blue fluorescence in apoptotic hepatocarcinoma cells,and the apoptosis of hepatocarcinoma cells increased with the increase of drug concentration.The percentage of cells in G0/G1 phase increased with the increase of cell cycle,and the ratio of cells in S phase was decreased in G2/M phase compared with blank control group（P〈0.05）;Western blot results showed that compared with the blank control group,TPh inhibited the proliferation of Bcl-2 cells in a concentration-dependent manner（P〈0.05）,and the number of apoptotic cells increased with the increase of drug concentration（P〈0.05）,and the ratio of Bcl-2/Bax in the TPh group decreased significantly（P〈0.05）,and the expression of Bax gene increased.Conclusion TPh inhibits cell proliferation,promotes apoptosis and induces HepG-2 to block G0/G1 phase.Its mechanism may increase the expression of Bax and decrease Bcl-2 protein expression.

关 键 词：TPH HEPG-2 细胞周期 细胞凋亡 BAX BCL-2 

分 类 号：R382.5[医药卫生—医学寄生虫学]