白杨素及其类似物BrMC对SMMC-7721细胞系球细胞DNMT1活性和球形成的影响

Effects of Chrysin and 8-bromo-7-methoxy chrysin on activity of DNMT1 and sphere forming capability of spheres from SMMC-7721 cell line

目的:检测白杨素(Chrysin,ChR)及其类似物8-溴-7-甲氧基白杨素(8-bromo-7-methoxy chrysin,BrMC)是否抑制SMMC-7721细胞系球细胞DNA甲基转移酶1(DNA Methyltransferase 1,DNMT1)活性和自我更新能力。方法:干细胞条件培养基悬浮培养法获得SMMC-7721细胞系球细胞;DNA甲基转移酶活性检测试剂盒测定DNMT1活性;球形成法检测肝癌球形成率;qRT-PCR分析干细胞表面标志物CD133和CD44 mRNA表达水平。结果:ChR(40μM)以及BrMC(5和10μM)显著抑制SMMC-7721细胞系球细胞DNMT1活性,降低肝癌球形成率,下调CD133和CD44 mRNA表达。结论:ChR和BrMC能有效地抑制SMMC-7721细胞系球细胞DNMT1活性和自我更新能力。

Objective To determine whether or not Chrysin(ChR)and its synthetic analogue 8-bromo-7-methoxy chrysin(BrMC)inhibit self-renewal capability and reduces of DNMT1 activity of spheres from SMMC-7721 cell line.Methods The spheres from SMMC-7721 cells were obtained by sphere-forming cultures.DNA methyltransferase activity assay kit was used to determine the activity of DNMT1.Sphere-forming assay was performed to examine sphere formation rate.Real-time quantitative PCR(qRT-PCR)was used to analyze the levels of CD133 and CD44 mRNAs.Results ChR(40μM)and BrMC(5 and 10μM)significantly repressed activity of DNMT1,reduced sphere formation rate,and downregulated expression of CD133 amd CD44 mRNAs.Conclusion ChR and BrMC can effectively repress DNMT1 activity and the self-renewal capacity of the spheres from SMMC-7721 cells.