过/降表达miR-92a的食管鳞癌细胞增殖、迁移、侵袭能力及PTEN/PI3K/Akt通路蛋白表达变化

Proliferation,migration,invasion and PTEN/PI3K/Akt pathway protein expression of esophageal squamous cell carcinoma cells with over-/down-expression of miR-92a

目的观察过/降表达miR-92a的食管鳞癌(ESCC)细胞增殖、迁移、侵袭能力及磷酸酯酶与张力蛋白同源物(PTEN)/磷脂酰肌醇3激酶(PI3K)/Akt通路蛋白表达变化。方法将人ESCC细胞系Eca-109随机分为miR-92a过表达组、miR-92a降表达组、过表达阴性对照组、降表达阴性对照组,分别转染miR-92a mimics、miR-92a inhibitor、mimic-NC、inhibitor-NC 24 h。采用CCK-8法检测各组培养24、48、72、96 h的的吸光度(A)值,划痕愈合实验检测细胞迁移能力,Transwell小室检测细胞侵袭能力。采用Westen blotting法检测细胞PTEN/PI3K/Akt通路蛋白PTEN、PI3K、Akt、磷酸化PTEN(p-PTEN)、p-PI3K、p-Akt。结果各时点细胞A值、细胞迁移距离、穿膜细胞数比较,miR-92a过表达组>过表达阴性对照组、降表达阴性对照组>miR-92a降表达组(P均<0.05)。p-PTEN蛋白miR-92a降表达组>过表达阴性对照组、降表达阴性对照组>miR-92a过表达组,p-PI3K、p-Akt蛋白miR-92a过表达组>过表达阴性对照组、降表达阴性对照组>miR-92a降表达组(P均<0.05),各组PTEN、PI3K、Akt蛋白差异均无统计学意义。结论miR-92a过/降表达可促进促进/抑制ESCC细胞的增殖、迁移及侵袭,其作用可能通过抑制/促进PTEN蛋白磷酸化调控PTEN/PI3K/Akt信号通路实现。

Objective To observe the proliferation,migration,invasion and protein expression of phosphatase and tensin homology(PTEN)/phosphatidylinositol 3 kinase(PI3K)/Akt pathway in esophageal squamous cell carcinoma(ESCC)cells with over-/down-expression of miR-92a.Methods Human ESCC cell line Eca-109 was randomly divided into the miR-92a over-expression group,miR-92a down-expression group,over-expression negative control group,and downexpression negative control group,which were transfected with miR-92a mimics,miR-92a inhibitor,mime-NC,and intrest-NC for 24 h,respectively.The absorbance(A)values of each group cultured for 24,48,72 and 96 h were detected by CCK-8.The cell migration ability was detected by scratch healing test,and the cell invasion ability was detected by Transwell chamber test.The protein expression levels of PTEN/PI3K/Akt pathway,PI3K,Akt,phosphorylated PTEN(pPTEN),p-PI3K,and p-Akt were detected by Western blotting.Results The comparison of cell A value,cell migration distance,and the number of transmembrane cells at each time point showed the following trend that miR-92a overexpression group>overexpression negative control group,and miR-92a down-expression negative control group>miR-92a downexpression group(all P<0.05).The p-PTEN protein expression of miR-92a was as follows:miR-92a down-expression group>overexpression negative control group,down-expression negative control group>miR-92a over-expression group,and the p-PI3K and p-Akt protein expression levels were as follows:miR-92a overexpression group>overexpression negative control group,down-expression negative control group>miR-92a down-expression group(all P<0.05).There were no significant differences in the expression of PTEN,PI3K,or Akt among all groups.Conclusion MiR-92a can promote/inhibit the proliferation,migration and invasion of ESCC cells,and its effect may be realized by inhibiting the phosphorylation of PTEN protein and regulating the PTEN/PI3K/Akt signaling pathway.