RNA干扰HDAC7表达在肝细胞癌中的作用

RNA interference of HDAC7 expression in hepatocellular carcinoma

目的:探讨组蛋白去乙酰化酶7(histone deacetylase 7,HDAC7)在肝细胞癌发生发展中的作用。方法:体外构建pSUPER-HDAC7反转录病毒干扰质粒,分为pSUPERHDAC7RNAi+组(有效干扰)、pSUPERHDAC7RNAi-组(无效干扰)和对照组(pSUPER空载体),转染人肝癌细胞HepG2和人微血管内皮细胞(human microvascular endothelial cell,HMEC-1)。Western免疫印迹检测HDAC7,p21,细胞周期素E(cyc-lin E)、基质金属蛋白酶10(matrix metalloproteinases 10,MMP10)、低氧诱导因子-1α(hypoxiainducible fac-tor-1α,HIF-1α)蛋白表达,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]、流式细胞术、体内裸鼠皮下种植、体外血管内皮细胞二维成管方法检测HDAC7表达的作用。结果:与pSUPERHDAC7RNAi-组和对照组相比,pSUPERHDAC7RNAi+组细胞HDAC7蛋白表达抑制,细胞生长抑制率和凋亡率明显增加,差异具有统计学意义(P<0.05);且HIF-1α和p21蛋白表达上调,cyclin E蛋白和MMP10表达下调,差异具有统计学意义(P<0.05)。结论:HDAC7蛋白通过上调p21和HIF-1α蛋白表达、下调cyclin E及MMP10蛋白表达,介导肝癌细胞凋亡和血管成管。

Objective To investigate the role of histone deacetylase 7（HDAC7） in the occurrence and development of hepatocellular carcinoma.Methods HepG2 cells and human microvascular endothelial cells（HMEC-1） were divided into 3 groups after transfection pSUPER-HDAC7 retroviral interference plasmid： a pSUPERHDAC7RNAi＋ group,a pSUPERHDAC7RNAi-group,and a pSUPER group as the control group.The expression of HDAC7,p21,cyclin E,matrix metalloproteinases 10（MMP10）,and hypoxiainducible factor-1α（HIF-1α） were detected by Western blot.The expression of HDAC7 in cell lines was determined by 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） and flow cytometry,the nude mice modle,and vascular endothelial cells 2-dimensional tubulogenesis in vitro and in vivo.Results Compared with the control group and the pSUPERHDAC7RNAi-group,the expression of HDAC7 was downregulated,the rate of cell growth inhibition and apoptosis in the pSUPERHDAC7RNAi＋ group increased more significantly;the expression of p21 and HIF-1α was increased significantly,while the expression of cyclin E and MMP10 in the pSUPERHDAC7RNAi＋ group was downregulated（P0.05）.Conclusion The expression of HDAC7 protein plays an important role in the apoptosis and vascular tubulogenesis of hepatocellular carcinoma by the upregulation of p21 and HIF-1α and the downregulation of cyclin E and MMP10.