ADH1B基因甲基化对卵巢癌细胞增殖及凋亡的影响  

Methylationof alcohol dehydrogenase 1B gene and its effects on cell proliferation and cell apoptosis of ovarian carcinoma cells

作　　者：季维雪 孙磊 江玉 陈颍[2] 李泽莲[2] 李敏[2] 肖兰 Ji Weixue;Sun Lei;Jiang Yu;Chen Ying;Li Zelian;Li Min;Xiao Lan(Dept of Obstetrics and Gynecology,Fuyang Hospital of Anhui Medical University,Fuyang 236112;Dept of Obstetrics and Gynecology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学附属阜阳医院妇产科,阜阳236112 [2]安徽医科大学第一附属医院妇产科,合肥230022

出　　处：《安徽医科大学学报》2022年第8期1274-1278,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学青年基金(编号:81603138);安徽省高校优秀拔尖人才培育项目(编号:gxgwfx2019006);安徽高校自然科学基金重大项目(编号:KJ2019ZD25)。

摘　　要：目的探讨乙醇脱氢酶1B(ADH1B)基因甲基化对卵巢癌(OC)细胞增殖及凋亡的影响。方法生物信息学方法比较ADH1B在卵巢癌与正常卵巢上皮组织中表达情况,实时荧光定量PCR(qRT-PCR)检测卵巢癌与正常卵巢组织中ADH1B相对表达量,对比两种研究结果的一致性。两株卵巢癌细胞OV2008及C13K经不同浓度甲基化抑制剂5-氮杂-2′-脱氧胞苷(5-Aza-dc)干预48 h,CCK-8法检测5-Aza-dc对两株细胞增殖的影响;Hoechst 33258染色检测细胞凋亡形态学改变;甲基化特异性PCR(MSP)、qRT-PCR及Western blot法分别检测药物作用前后两株细胞株中ADH1B甲基化状态、ADH1B在mRNA及蛋白水平表达。结果生物信息学分析及qRT-PCR结果均显示卵巢癌中ADH1B表达量明显均低于正常卵巢组织;中、高浓度5-Aza-dc作用48 h后,两株细胞增殖受到抑制(P<0.01);高浓度5-Aza-dc作用后,两株细胞均出现典型细胞凋亡形态学改变;ADH1B在两株细胞中均呈完全甲基化,高浓度5-Aza-dc处理后,ADH1B甲基化被部分逆转,两株细胞ADH1B mRNA和蛋白表达均增加(P<0.01)。结论5-Aza-dc可下调卵巢癌细胞中ADH1B启动子区甲基化水平,使ADH1B基因重新表达,从而抑制卵巢癌细胞增殖与促进细胞凋亡。Objective To investigate the effects of methylation expression of ADH1B gene on cell proliferation and cell apoptosis in ovarian cancer(OC)cells.Methods Bioinformatics analysis was performed to compare the relative expression of ADH1B in OC and normal tissue.Quantitative real-time PCR(qRT-PCR)was used to detect the ADH1B mRNA expression in the OC tissues or cells,ovary normal epithelium tissues or cells.Treated with different concentrations of 5-Aza-dc for 48 h,cell viability was determined by CCK-8 method.Cell apoptosis morphology was detected by Hoechst 33258 staining.The changes of promoter methylation of ADH1B,the level of ADH1B in the mRNA and protein expressions were measured by using methylation specific PCR(MSP),qRT-PCR and Western blot,respectively.Results Bioinformatics analysis and qRT-PCR showed that ADH1B expression in OC was lower than that in normal tissue.After treatment with medium and high concentration of 5-Aza-dc,the cell proliferation was inhibited in two cells(P<0.01).After treatment with high concentration of 5-Aza-dc,marked morphological changes of apoptosis was observed in two cells.ADH1B was completely methylated in two cells.ADH1B was partially reversed by high concentration of 5-Aza-dC,and ADH1B mRNA and protein expression both increased by 5-Aza-dC in two cells(P<0.01).Conclusion 5-Aza-dC can down-regulate the methylation level of tumor suppressor gene ADH1B promoter region and make it re-express in ovarian cancer cells,thus exert its function of enhancing the inhibitory effect and apoptosis of ovarian cancer cells.

关 键 词：乙醇脱氢酶1B 甲基化 5-氮杂-2′-脱氧胞苷 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]