Objective To study the effect of hepatitis B vin1s X-interacting protein (HBXIP) on the proliferation,migration,and invasion of ACC-2 cells,and the possible mechanism of the PI3K/Akt signaling pathway involved.Methods The chemically synthesized HBXIP-siRNA plasmid was transfected into the ACC-2 cells.RT-PCR and Western blotting were performed to detect the expression of HBXIP in the ACC-2 cells.Cell proliferation was measured via MTT assay.The invasive and migratory abilities of the ACC-2 cells were evaluated via the transwell chamber assay and scratch test,respectively.Western blotting also detected the impact of HBXIP-siRNA on Akt,p-Akt,PI3K,p-PI3K,and S100A4 protein expression.Results HBXIP was highly expressed and HBXIP-siRNA was successfully transfected in ACC-2 cells.MTT results showed that the number of surviving cells in the experimental group was significantly lower than that in the control group (P＜0.05).The scratch test results showed that the mobility of the experimental group was significantly lower than that of the control group (P＜0.01).The transwell assay showed that the rate of cell invasion of the experimental group was significantly lower than that of the control group (P＜0.01).Finally,Western blotting results revealed that the expression of p-Akt,p-PI3K,and S 100A4 was relatively decreased in the experimental group when compared to that in the control group.Conclusion Silencing the HBXIP gene inhibited ACC-2 proliferation,invasion,and migration.%目的 探讨乙肝病毒X蛋白结合蛋白(HBXIP)对腺样囊性癌细胞株ACC-2生物学行为的影响及其机制.方法 采用siRNA转染法抑制腺样囊性癌细胞株ACC-2中HBXIP的表达;实时PCR和Western blotting检测HBXIP基因及蛋白表达水平;MTT法检测HBXIP-siRNA对细胞增殖的影响;transwell法检测HBXIP-siRNA对细胞侵袭的影响;划痕实验检测HBXIP-siRNA对细胞迁移的影响;蛋白印迹方法检测HBXIP-siRNA对PI3K/Akt信号通路中Akt、p-Akt、PI3K、p-PI3K及对S100A4蛋白表达量的影响.结果 HBXIP在ACC-2中高表达,HBXIP-siRNA成功转染细胞株.MTF结果显示,48、72、96 h时实验组中存活的细胞数低于对照组,差异有统计学意义(P＜0.05);划痕实验结果显示,实验组细胞迁移率明显低于对照组(P＜0.01);transwell小室实验结果显示,实验组中细胞侵袭个数明显低于对照组(P＜0.01);蛋白印迹结果显示,相对于对照组,实验组细胞中p-Akt、p-PI3K及S100A4的表达量随着HBXIP基因沉默而相对降低.结论 HBXIP可促进腺样囊性癌细胞株ACC-2增殖、迁移和侵袭,其机制可能与促进Akt、PI3K磷酸化及增加S100A4蛋白表达量有关.
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