茯苓酸抑制TRIM29表达通过Wnt信号通路调控宫颈癌细胞存活和凋亡

Porialic Acid Regulated and Controlled Survival and Apoptosis in Cervical Cancer Cells through Wnt Signaling Pathway by Inhibiting TRIM29 Expression

【目的】观察茯苓酸对宫颈癌细胞Caski存活、凋亡的影响及潜在作用机制。【方法】采用四甲基偶氮唑盐(MTT)法观察茯苓酸处理或转染的Caski细胞生存能力,流式细胞术观察茯苓酸处理或转染的Caski细胞凋亡情况,实时定量逆转录聚合酶链反应(RT-qPCR)法检测茯苓酸处理或转染的Caski细胞三基序蛋白29(TRIM29)mRNA的表达,蛋白免疫印迹(Western Blot)法检测茯苓酸处理或转染的Caski细胞TRIM29、含半胱氨酸的天冬氨酸蛋白水解酶9(Caspase-9)、细胞周期调节蛋白Cyclin D1及Wnt通路蛋白β-连环蛋白(β-catenin)、糖原合成酶激酶3β(GSK-3β)、c-Myc的表达。【结果】茯苓酸能够降低Caski细胞存活率,促进细胞凋亡,升高Caspase-9、β-catenin表达量,降低Cyclin D1、GSK-3β、c-Myc表达量,抑制TRIM29的mRNA和蛋白表达。沉默TRIM29可降低Caski细胞存活率,促进细胞凋亡,促进Caspase-9表达,抑制Cyclin D1表达。过表达TRIM29可通过上调Wnt通路活性部分逆转茯苓酸对宫颈癌Caski细胞存活和凋亡的作用。【结论】茯苓酸通过抑制TRIM29表达下调Wnt通路活性从而抑制宫颈癌Caski细胞存活,促进细胞凋亡。

Objective To observe the effect of pachymic acid(PA)on the survival and apoptosis in cervical cancer cells Caski and its potential mechanism.Methods The survival of Caski cells treated with PA and/or transfected was observed by MTT assay,the apoptosis of Caski cells treated with PA and/or transfected was observed by flow cytometry,the mRNA expression level of tripartite motif protein 29(TRIM29)in Caski cells treated with PA and/or transfected was detected by real time quantitative reverse transcription polymerase chain reaction(RT-qPCR),and the expression levels of TRIM29,Caspase-9,cell cycle-regulatory protein Cyclin D1,and Wnt signaling pathway proteinsβ-catenin,glycogen synthase kinase 3β(GSK-3β),c-Myc in Caski cells treated with PA and/or transfected were detected by Western blotting method.Results PA decreased Caski cell survival rate,promoted Caski cell apoptosis,increased the expression quantity of Caspase-9 andβ-catenin in Caski cells,decreased the expression quantity of cyclin D1,GSK-3βand c-Myc,inhibited the mRNA and protein expression of TRIM29.Silencing TRIM29 decreased Caski cell survival rate,promoted cell apoptosis,enhanced Caspase-9 expression,and inhibited Cyclin D1 expression.Overexpression of TRIM29 partially reversed the effect of PA on survival and apoptosis in Caski cells by up-regulating the activity of Wnt signaling pathway.Conclusion PA inhibits the survival of cervical cancer Caski cells and promotes cell apoptosis through down-regulating Wnt pathway activity by inhibiting TRIM29.