二甲双胍对人绒癌JAR细胞上皮-间质转化和侵袭迁移能力的影响

Effects of metformin on epithelial-mesenchymal transition,migration and invasion in human choriocarcinoma cell line JAR

目的 探索二甲双胍对人绒癌JAR细胞株上皮-间质转化,侵袭和迁移能力的影响。方法 使用浓度为40 mmol/L的二甲双胍处理人绒癌JAR细胞株,随后分别使用基于Transwell小室的实时荧光照相技术和划痕实验分别检测二甲双胍处理前后JAR细胞的侵袭和迁移变化;使用实时荧光定量聚合酶链反应和免疫蛋白印记法检测可能调控上述现象的分子:腺苷酸活化蛋白激酶(AMPK)、磷酸化腺苷酸活化蛋白激酶( p-AMPK )、上皮细胞钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)的mRNA及蛋白变化趋势。结果 二甲双胍处理人绒癌JAR细胞株24 h后AMPK的mRNA表达增加,AMPK总蛋白无明显变化而p-AMPK蛋白表达上调。细胞的迁移和侵袭能力减弱,同时上皮-间质转化分子标志物E-cadherin上调,N-cadherin和Vimentin下调,提示上皮-间质转化能力降低。结论 二甲双胍可以通过磷酸化激活AMPK抑制人绒癌JAR细胞株的上皮-间质转化,侵袭和迁移能力。

Objective To investigate the effects of metformin on epithelial-mesenchymal transition (EMT),migration and invasion in human choriocarcinoma cell line JAR. Methods The human choriocarcinoma cell line JAR was divided into control and metformin treated (40 mmol/L) groups.Cell migration and invasion ability were determined by the wound healing assay and fluorescent transwell chamber assay.The expression of AMPK,phosphorylated AMPK and EMT-related markers:E-cadherin,N-cadherin and Vimentin were measured by real-time PCR and Western Blot respectively. Results The mRNA expression of AMPK was increased,but total AMPK protein was unchanged.However,phosphorylated AMPK (p-AMPK) protein was significantly increased in metformin group compared with control group.The ability of cell migration and invasion were decelerated in metformin group.Epithelial marker E-cadherin was up-regulated and mesenchymal markers N-cadherin and Vimentin were down-regulated after treated with metformin. Conclusion AMPK activation may inhibit the EMT,migration and invasion in human choriocarcinoma cell line JAR.