DNA甲基化转移酶抑制剂5-aza-dC通过上调WIF-1表达抑制子宫内膜癌细胞增殖并诱导凋亡的作用机制研究  

DNA methyltransferase inhibitor 5-aza-dC inhibits endometrial cancer cell proliferation and induces apoptosis through up regulation of WIF-1 expression

作　　者：张保华[1] 周小莉 季静[1] 胡明珠 李岚[1] 聂启勤 蓝波 杨慧宇 徐红 ZHANG Bao-hua;ZHOU Xiao-li;JI Jing;HU Ming-zhu;LI Lan;NIE Qi-qin;LAN Bo;YANG Hui-yu;XU Hong(Department of Obstetrics and Gynecology,Punan Hopitail,Shangai Pudong New District,Shanghai 200125,China)

机构地区：[1]上海市浦东新区浦南医院,妇产科,上海200125

出　　处：《医药论坛杂志》2022年第20期6-11,16,共7页Journal of Medical Forum

基　　金：浦东新区卫生和计划生育委员会领先人才培养(PWRl2018-10);浦东新区卫生健康委员会卫生计生科研项目(PW2019D-5)。

摘　　要：目的 探讨5-aza-dC通过上调WIF-1基因表达对子宫内膜癌细胞增殖和凋亡的影响并阐明作用机制。方法 通过慢病毒途径在子宫内膜癌细胞JEC和KLE中靶向沉默WIF-1基因。使用梯度5-aza-dC处理WIF-1基因沉默及未沉默的子宫内膜癌细胞分别检测细胞增殖活性及凋亡率,同时检测WIF-1及β-catenin蛋白表达或者磷酸化。结果 通过慢病毒途径,可以在JEC和KLE细胞中有效沉默WIF-1基因(P<0.01,vs细胞对照组)。1和5μM的5-aza-dC能够有效抑制JEC细胞对数期增殖活性(P<0.01,vs细胞对照组或者溶媒对照组72 h)。1和5μM的5-aza-dC能够有效诱导KLE细胞发生凋亡(P<0.01,vs细胞对照组或者溶媒对照组)。免疫印迹检测结果表明,5μM的5-aza-dC能够有效上调JEC和KLE细胞中WIF-1表达并下调β-catenin蛋白磷酸化。5-aza-dC的上述作用均呈现梯度依赖,且能够被WIF-1沉默所逆转(P<0.01,vs 5-aza-dC处理组)。结论 5-aza-dC能够通过抑制Wnt/β-catenin通路活化而抑制子宫内膜癌细胞的增殖并诱导凋亡,且作用依赖于其对WIF-1的表达上调。Objective To clarify the effect of 5-aza-dC on the proliferation and apoptosis of endometrial cancer cells by up-regulating the expression of WIF-1 gene, and to clarify the mechanism.Methods WIF-1 gene was silenced in endometrial cancer cells JEC and KLE by lentivirus.Endometrial cancer cells with WIF-1 silenced and non silenced were treated with gradient 5-aza-dC.Then the cell proliferation activity, apoptosis rate, expression of WIF-1 and phosphorylated β-catenin were detected.Results WIF-1 gene was effectively silenced in JEC and KLE cells by lentivirus(P<0.01, vs cell control group).1 and 5 μM 5-aza-dC could effectively inhibit the logarithmic proliferation of JEC cells(P<0.01, vs cell control group or vehicle control group.72 h).1 and 5 μM 5-aza-dC can effectively induce the apoptosis of KLE cells(P<0.01, vs cell control group or vehicle control group).Western blot results showed that 5μM 5-aza-dC can effectively up-regulate the expression of WIF-1 and down-regulate the phosphorylation of β-catenin in JEC and KLE cells.The above effects of 5-aza-dC were gradient dependent and could be reversed by WIF-1 silencing(P<0.01, vs 5-aza-dC treatment group).Conclusion 5-aza-dC can inhibit the proliferation and induces apoptosis of endometrial cancer cells by suppressing the Wnt/β-catenin pathway, and its effect depends on the up-regulation of WIF-1.

关 键 词：5-AZA-DC 子宫内膜癌 WIF-1 WNT/Β-CATENIN 

分 类 号：R711.74[医药卫生—妇产科学]