摘要
目的检测DNA甲基转移酶(DNMTs)抑制剂5-氮杂-2'脱氧胞苷(5-Aza-2'deoxycytidine,5-Aza-CdR)处理后,对宫颈癌细胞系Hela、Siha、C33A的增殖、HPV16、HPV18E6 mRNA、TERT mRNA水平以及对于p21、p53蛋白表达的影响。方法用不同浓度5-Aza-CdR干预宫颈癌Hela、Siha、C33A细胞株7 d后,采用CCK8检测三种细胞增殖水平的变化;RT-PCR检测HPV16、HPV18 E6以及TERT mRNA水平;Western blot法检测p21和p53蛋白表达。结果随着5-Aza-CdR浓度的增加,Hela、Siha、C33A细胞的增殖抑制随之增强,呈时间剂量依赖关系。5-Aza-CdR对于C33A、Hela、Siha、细胞抑制率分别为70.3%、61.9%、40.6%,差异有统计学意义(P <0.05);去甲基化后,Siha细胞HPV16E6和Hela细胞HPV18 E6水平呈下降趋势,Hela、Siha、C33A细胞TERT mRNA水平均呈下降趋势;去甲基化后宫颈癌Siha、Hela和C33A细胞中p53和p21蛋白含量明显增加,差异有统计学意义(P <0.05)。结论 5-Aza-CdR可能通过激活p53、p21信号通路抑制TERT mRNA水平,进而通过抑制HPVE6水平抑制宫颈癌细胞的增殖。
Objective After treatment with inhibitors of DNA methyltransferase(DNMTs) 5-Aza-CdR,the proliferation of Hela,Siha,C33 A,the expression of HPV16,HPV18 E6,TERT mRNA levels of cervical cancer cell lines and the expression of p21 and p53 proteins were observed.Methods After intervention with different concentrations of 5-Aza-CdR in cervical cancer Hela,Siha and C33 A cell lines for 7 days,the proliferation changes of the three kinds of cells were detected by CCK8.Changes in HPV16,HPV18 E6 mRNA and TERT mRNA expression were detected by RT-PCR.Protein expression of p21 and p53 was detected by Western blot.Results With the increase of 5-Aza-CdR concentration,the proliferation inhibition of Hela,Siha and C33 A cells increased,showing a time and dose-dependent relationship.The inhibition rates of C33 A,Hela and Siha cells were 70.3%,61.9% and 40.6%,respectively,with statistically signif icant difference(P <0.05).After demethylation,the levels of HPV16 E6 in Siha cells and HPV18 E6 in Hela cells showed a downward trend,while the levels of TERT mRNA in Hela,Siha and C33 A cells all showed a downward trend.After demethylation,p53 and p21 protein contents in Siha,Hela and C33 A cells of cervical cancer were signif icantly increased(P <0.05).Conclusions 5-Aza-CdR can inhibit TERT mRNA levels by activating p53 and p21 signaling pathways,thereby inhibiting HPVE6 levels and inhibiting cervical cancer cell growth.
作者
李明珠
李换男
赵丽君
王甲琪
魏丽惠
LI Mingzhu;LI Huannan;ZHAO Lijun;WANG Jiaqi;WEI Lihui(Peking University People Hospital. Beijing 100044, China)
出处
《中国妇产科临床杂志》
CSCD
北大核心
2019年第2期153-156,共4页
Chinese Journal of Clinical Obstetrics and Gynecology
基金
北京市自然科学基金(7174359)
国家重点研发计划(2016YFC1302901)
国家科技支撑计划(2015BAI13B06)
