卵巢癌细胞PI3K/AKT通路与肾上腺髓质素促迁移作用的相关性研究
Function of AKT Phosphorylation in AM Promotion of HO8910 Migration
-
摘要:目的 检测PI3K/AKT信号通路在肾上腺髓质素(AM)促卵巢癌细胞HO8910迁移中的作用。方法 利用划痕实验检测外源性AM对卵巢癌细胞HO8910迁移功能的影响, 并且检测PI3K抑制剂Wortmannin对AM促进卵巢癌细胞迁移功能的干扰。为进一步研究AM对AKT磷酸化的影响, 应用蛋白印迹法检测细胞AKT蛋白磷酸化的表达, 应用整合素α5β1抑制性抗体(mAb)拮抗AM对卵巢癌细胞AKT磷酸化作用。结果 外源性AM处理的卵巢癌细胞12 h迁移率(62.83±4.46)%明显高于正常卵巢癌细胞的迁移率(30.26±1.55)%(P < 0.001), PI3K抑制剂Wortmannin干扰1 h后再用Am处理12 h, 细胞迁移率为(40.44±0.88)%, 明显低于单纯应用AM刺激的细胞(P=0.001)。AM可以促进卵巢癌细胞迁移(P < 0.001), PI3K抑制剂Wortmannin可以拮抗AM的促迁移作用(P=0.001)。AM可以促进细胞AKT蛋白磷酸化, Wortmannin、整合素α5β1抑制性抗体均可以干扰AM对AKT蛋白磷酸化作用。结论 在卵巢癌HO8910细胞中, PI3K/AKT的活化是AM促进卵巢癌细胞迁移重要机制, 而整合素α5β1可能作为感受器参与AM促进细胞PI3K/AKT通路的活化。Abstract:Objective This study aims to investigate the fimction of PI3K / AKT in the adrenomedullin (AM) promotion of ovarian cancer cell migration.Methods AM and Wortmannin, as inhibitors of PI3K /AKT, were used to impede AM function on cell migration. Wound healing test was performed to illustrate the migration ability of ovarian cancer cells. Western blot analysis was performed to analyze AKT protein levels and its phosphorylation pattern in cells after providing AM, Wortmannin, and integrin α5β1 blocking antibody.Results AM significantly improved the migration of the ovarian cancer cells (P < 0.001). Wortmannin inhibited AM function in HO8910 cell migration (P < 0.001). AKT phosporylation in cells was significantly increased at 15 min after the addition of AM. Wortmannin and integrin α5β1 blocking antibody reduced AKT phosphorylation by AM.Conclusions AM induces ovarian epithelial cancer cell migration by phosphorylating PI3K/AKT via integrin α5β1 activation.
下载: