MBD2过表达对子宫肌瘤生物学功能的影响及其基因启动子区甲基化分析  

Overexpression of MBD2 on biological function of uterine leiomyoma cell and promoter methylation of MBD2 in human uterine leiomyoma

作　　者：梁美蓉[1,2] 曾泱 曾四元[1,2] 张俊玟 邹阳 黄欧平 LIANG Meirong;ZENG Yang;ZENG Siyuan;ZHANG Junwen;ZOU Yang;HUANG Ouping(Medical College of Nanchang University,Nanchang,Jiangxi 330006,China)

机构地区：[1]南昌大学医学院,330006 [2]江西省妇幼保健院肿瘤科 [3]江西省妇幼保健院妇科 [4]江西省女性生殖健康重点实验室

出　　处：《中国妇产科临床杂志》2019年第6期519-522,共4页Chinese Journal of Clinical Obstetrics and Gynecology

基　　金：江西省科技厅应用研究培育计划（20181BBG78016）;江西省卫生计生委普通科技计划（20185391）

摘　　要：目的前期研究发现子宫肌瘤组织中存在甲基化CpG结合蛋白2(methyl-CpG binding domain protein 2,MBD2)基因和蛋白的差异性表达下调,本研究探讨MBD2过表达对子宫肌瘤生物学功能的影响,并分析子宫肌瘤组织中MBD2基因启动子区甲基化状态。方法采用慢病毒包装MBD2真核过表达载体,通过转染至293T细胞,检测293T细胞的增殖、凋亡及周期情况;利用飞行时间质谱技术检测子宫肌瘤(病例组)及瘤旁肌层组织(对照组)中MBD2的基因启动子区甲基化水平,分析MBD2基因CpG位点的甲基化率与MBD2mRNA水平的关系。结果①MBD2真核过表达载体成功转染293T细胞,上调293T细胞MBD2的表达对其增殖、凋亡和周期均无显著影响。②共46对标本的MBD2基因启动子区的10个CpG位点进行甲基化分析法发现,单点甲基化水平分析中,所有CpG位点上,MBD2 mRNA与MBD2甲基化的表达在病例组与对照组比较,差异均无统计学意义(P均>0.05)。结论过表达MBD2对子宫肌瘤细胞生物学功能并不产生影响,子宫肌瘤组织中MBD2基因的表达下调与其基因启动子区甲基化状可能无关。Objective In this study,we explored the effects of MBD2 overexpressionon biological function of uterine leiomyoma cell and promoter methylation of MBD2 in human uterine leiomyoma(UL)based on our previous results that both the mRNA and protein expression of MBD2 significantly decreased in ULs compared to adjacent myometrium.Methods To over-expression MBD2,lentiviral vectors expressing MBD2(pLV[Exp]-Puro-CMV>hMBD2)were transfected into 293 T cell lines,and then the proliferation,cell cycle and apoptosis were observed.Sequenom Mass Array platform was performed to examine the DNA methylation level of MBD2 in UL and paired adjacent myometrium tissues,and to explore the potential association between MBD2 differential expression and the DNA methylation status changes in MBD2 gene promoter region.Results pLV[Exp]-Puro-CMV>hMBD2 were transfected into 293T cell lines successfully,however,over-expression MBD2 had no effect on proliferation,cell cycle and apoptosis of 293T cell lines.Furthermore,the methylation status of a total of 10 CpG sites within the MBD2 gene promoter in 46 UL tissues and paired adjacent myometrium were analyzed.The expression of MBD2 mRNA and MBD2 methylation at all CpG sites was compared between the case group and the control group.The differences were not statistically significant.Conclusions Overexpression of MBD2 has no effects on the biological function of uterine leiomyoma cell.Furthermore,the DNA methylation status in the promoter region of the MBD2 gene might have no correlation with the decreased expression of MBD2 in UL tissues.

关 键 词：子宫肌瘤 发病机制 MBD2 DNA甲基化 

分 类 号：R73[医药卫生—肿瘤]