异甘草素通过调控PI3K/AKT/mTOR通路对子宫内膜癌细胞增殖、迁移的影响

Effects of isoliquiritigenin on proliferation and migration of endometrial cancer cells by regulating PI3K/AKT/mTOR pathway

目的探讨异甘草素调控磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路对子宫内膜癌细胞增殖、迁移的影响。方法以MTT法筛选异甘草素最佳作用浓度。体外培养Ishikawa细胞并构建其移植瘤裸鼠模型,均随机分为对照组、异甘草素组、740 Y-P组(PI3K激活剂)、异甘草素+740 Y-P组,均以异甘草素和740Y-P分组处理细胞和裸鼠后,采用MTT法和平板集落形成实验检测各组细胞增殖;采用TUNEL染色检测各组细胞凋亡;采用划痕实验和Transwell实验检测各组细胞迁移;检测各组裸鼠移植瘤体积与质量;采用免疫印迹法检测各组细胞凋亡相关蛋白、上皮间充质转化相关蛋白和PI3K/AKT/mTOR通路相关蛋白表达。结果与对照组相比,异甘草素组细胞活力、集落形成率、迁移率、迁移数、移植瘤体积与质量、Bcl-2、纤维连接蛋白(FN)及Vimentin蛋白表达、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR降低(P<0.05),细胞凋亡率、Bax及E-cadherin蛋白表达升高(P<0.05);740 Y-P组各指标变化趋势与异甘草素组相反。740 Y-P可减弱异甘草素对Ishikawa细胞各指标的作用。结论异甘草素可抑制PI3K/AKT/mTOR通路蛋白磷酸化,进而抑制子宫内膜癌细胞增殖、集落形成、迁移及裸鼠体内移植瘤生长,并增强子宫内膜癌细胞凋亡。

Objective To investigate the effects of isoliquiritigenin on proliferation and migration of endometrial cancer cells by regulating phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway.Methods The optimal concentration of isoliquiritigenin was selected by MTT method.Ishikawa cells were cultured in vitro and their transplanted tumor models were constructed in nude mice,and they were randomly grouped into a control group,an isoliquiritigenin group,a 740 Y-P group(PI3K activator),and an isoliquiritigenin+740 Y-P group,after cells and nude mice were treated with isoglycyrrhizin and 740 Y-P,MTT assay and plate colony formation assay were applied to detect cell proliferation in each group.TUNEL staining was applied to detect cell apoptosis in each group.Scratch test and Transwell test were applied to detect cell migration in each group.The volume and weight of transplanted tumors of nude mice in each group were detected.Immunoblotting was applied to detect the expression of apoptosis related proteins,epithelial mesenchymal transition related proteins,and PI3K/AKT/mTOR pathway related proteins in each group.Results Compared with the control group,the cell viability,colony formation rate,migration rate,migration number,transplanted tumor volume and weight,Bcl-2,fibronectin(FN),and Vimentin protein expression,p-PI3K/PI3K,p-AKT/AKT and p-mTOR/mTOR in the isoliquiritigenin group reduced(P<0.05),the apoptosis rate,Bax and E-cadherin protein expression increased(P<0.05).The trend of changes in various indicators in the 740 Y-P group is opposite to that in the isoliquiritigenin group.740 Y-P can weaken the effect of isoliquiritigenin on various indicators of Ishikawa cells.Conclusion Isoglycyrrhizin can inhibit the phosphorylation of PI3K/AKT/mTOR pathway proteins,thereby inhibiting endometrial cancer cell proliferation,colony formation,enhancing its apoptosis and tumor growth and migration in nude mice,and enhancing endometrial cancer cell apoptosis.