己烯雌酚对OVCAR3细胞株HOXA10基因表达与甲基化影响及其机制的探讨  

Effect of diethylstilbestrol on the expression and methylation of HOXA10 gene in OVCAR3 cell line

作　　者：万一聪[1] 张林[1] 姜旖[1] 马小玲[1] 程文俊[1] 

机构地区：[1]南京医科大学第一附属医院妇科,江苏南京210029

出　　处：《中华肿瘤防治杂志》2012年第22期1681-1684,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金面上项目(30772331);江苏省科技厅自然科学基金面上项目(BK2010576);江苏省六大人才高峰(303070774IB09)

摘　　要：目的:研究己烯雌酚(DES)对卵巢上皮性癌细胞株OVCAR3中HOXA10基因表达及启动子区甲基化状态的影响。方法:体外培养卵巢上皮性癌细胞株OVCAR3,分别加入5×10-6、5×10-7和5×10-8 mol/L的DES培养5d(120h)后收集细胞,采用实时荧光定量聚合酶链反应和蛋白免疫印记技术检测HOXA10基因mRNA和蛋白的表达水平;运用甲基化特异性聚合酶链反应检测HOXA10基因启动子区的甲基化状态。结果:随着DES浓度的升高,OVCAR3细胞中HOXA10基因的表达存在升高的趋势,且各组间的表达差异有统计学意义,HOXA10基因mRNA表达(5×10-8 mol/L DES组与对照组比较,P=0.002;5×10-7 mol/L DES组与5×10-8 mol/L DES组比较,P=0.028;5×10-6 mol/L DES组与5×10-7 mol/L DES组比较,P=0.000 2)、HOXA10基因蛋白表达(5×10-8 mol/L DES组与对照组比较,P=0.032;5×10-7 mol/L DES组与5×10-8 mol/L DES组比较,P=0.049;5×10-6 mol/L DES组与5×10-7 mol/LDES组比较,P=0.009);且DES可诱导HOXA10基因启动子区的甲基化状态发生去甲基化改变。结论:DES可以上调卵巢上皮性癌细胞株OVCAR3细胞中HOXA10基因的表达,这与DES对HOXA10基因启动子区甲基化状态的影响有关。OBJECTIVE： To investigate the effect of diethylstilbestrol （DES） on the expression and promoter region methylation status of HOXA10 gene in ovarian epithelial cancer cell line OVCAR3. METHODS： The ovarian epithelial cancer cell line OVCAR3 was incubated with 5Х10 6,5Х10^-7 and 5Х10^-8mol /L DES for 5 d （120 h）.Then Real Time-PCR and Western blot were used to detect the expression of HOXA10 mRNA and protein and methylation specific polymerase chain reaction （MSP） to detect HOXA10 gene promoter region methylation status. RESULTS： With the in- creased DES concentration, HOXA10 gene expression was elevated in the OVCAR3 cell line, and there was significant difference between each group, HOXA10 mRNA level （5 X 10 8 mol/L vs control,P=0. 002; 5 X 10 7 mol/L vs 5 X 10^-8 mol/L,P^0.028; 5Х10-6 mol/LvsS^10 7 mol/L,P=0.0002）,HOXA10proteinlevel （5Х10^-8 mol/Lvscontrol,P=0. 032; 5Х 10^-7 mol/L vs 5Х 10^-8 mol/L,P=0.04g; 5 X 10-6 mol/L vs 5 X 10^-7 mol/L,P=0. 009）,and after exposure to DES the methylation status of HOXA10 gene promoter region changed. CONCLUSION： DES could upregulate HOXA10 expression in ovarian epithelial cancer cell line OVCAR3,which related to the change in the methylation status of HOXA10 gene promoter region induced by DES.

关 键 词：卵巢肿瘤 己烯雌酚 同源盒基因 DNA甲基化 

分 类 号：R737.31[医药卫生—肿瘤]