RNA干扰HDAC7表达在肝细胞癌中的作用

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(cyclin E)、基质金属蛋白酶10（matrix metalloproteinases 10, MMP10）、低氧诱导因子-1α（hypoxiainducible factor-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-dimethylthiazol-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。

RNA interference of HDAC7 expression in hepatocellular carcinoma

ObjectiveTo investigate the role of histone deacetylase 7 (HDAC7) in the occurrence and development of hepatocellular carcinoma. MethodsHepG2 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. ResultsCompared 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 (P＜0.05). ConclusionThe 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.