索拉非尼联合5-氮杂-2'-脱氧胞苷对肝癌SMMC-7721细胞DNMT3B基因表达的影响

Effect of Sorafenib combined with 5-nitrogen impurity-2'-deoxy cytidine on the expression of DNMT3B gene in hepatocellular carcinoma SMMC-7721 cells

目的:探讨索拉非尼联合5-氮杂-2'-脱氧胞苷(5-Aza-CdR)对人肝癌细胞SMMC-7721的作用及其调控DNMT3B基因表达可能的分子机制。方法:单独及联合给药后以MTT法测定SMMC-7721的增殖,Transwell检测其侵袭,流式细胞仪检测细胞凋亡,Western blot法观察DNMT3B、p-Akt-Ser473及ERK蛋白的表达。结果:索拉非尼、5-Aza-CdR对肝癌SMMC-7721细胞半数抑制浓度(IC50)分别为11、6μmol/L,联合用药选取(6+4)μmol/L作为后续用药浓度。与对照组比较单药及联合用药均能抑制细胞侵袭、促进细胞凋亡、减少p-Akt-Ser473和DNMT3B蛋白的表达(P<0.05~P<0.01)。结论:索拉非尼和5-AzaCdR联合用药能有效降低索拉非尼用药浓度,降低肝癌SMMC-7721细胞DNMT3B蛋白表达水平。

Objective： To study the effects of the sorafenib combined with 5-nitrogen impurity-2＇-deoxy cytidine on human hepatocellular carcinoma SMMC-7721 cells, and investigate the possible molecular mechanism of regulating the DNMT3B gene expression. Methods： After separate and combined medication, the proliferation, invasion and apoptosis of SMMC-7721 ceils were detected using MTr method, transwell method and flow cytometry, respectively. The protein expressions of DNMT3B, p-Akt -Ser473 and ERK were measured using Western blotting. Results：The IC50 of sorafenib and 5-nitrogen impurity-2＇-deoxy cytidine to hepatocellular carcinoma SMMC-7721 cells were 11 μmol/L and 6 μmol/L respectively. The 6 μmol/L of sorafenib combined with 4 μmol/L of 5-nitrogen impurity-2＇-deoxy cytidine was used as the subsequent drug concentration. Compared with the control group, the separate and combined medication could inhibit the cell invasion, promote apoptosis and reduce the protein expressions of p-Akt-Ser473 and DNMT3 B （P 〈 0.05 to P 〈 0.01 ）. Conclusions：The combined medication of Sorafenib with 5-nitrogen impurity-2＇-deoxy cytidine can effectively reduce the sorafenib drug concentration and the protein expression levels of DNMT3B in hepatocellular carcinoma SMMC- 7721 cells.