HBXIP对腺样囊性癌细胞株ACC-M生物学功能及PI3K/Akt信号通路的影响

目的:研究乙肝病毒X蛋白结合蛋白(hepatitis B X-interacting protein,HBXIP)对唾液腺腺样囊性癌肺高转移细胞株(ACC-M)增殖、迁移和侵袭的影响,及其对PI3K/Akt信号通路的影响.方法:将HBXIP质粒转染到ACC-M中,将细胞分为实验组(转染pEGFP-N1-HBXIP质粒)、对照组1(未转染组)和对照组2(vector组,pEGFP-N1).采用RT-PCR检测HBXIP在ACC-M中的表达;采用MTT实验、Transwell小室实验和划痕实验检测HBXIP过表达对ACC-M的增殖、迁移和侵袭的影响;Western印迹法检测HBXIP过表达对Akt、p-Akt、PI3K、p-PI3K及S100A4蛋白表达量的影响.采用SPSS18.0软件包对数据进行统计学分析.结果:MTT实验结果显示,实验组中存活的细胞数显著高于对照组(P＜0.05);划痕实验结果显示,实验组细胞迁移率显著高于对照组(P＜0.01);Transwell小室实验结果显示,实验组细胞侵袭个数显著高于对照组(P＜0.01);Western印迹法结果显示,相对于对照组,实验组随着HBXIP过表达,p-Akt、p-PI3K及S100A4表达量相对增高.结论:HBXIP基因过表达ACC-M增殖、迁移和侵袭具有影响,可能通过促进Akt、PI3K磷酸化及S100A4蛋白表达量增加,促进ACC-M的增殖、迁移和侵袭.

Effect of HBXIP on biological function and PI3K/Akt signaling pathway of adenoid cystic carcinoma cell line ACC-M

PURPOSE:To study the effect of hepatitis B virus X protein binding protein (HBXIP) on proliferation,migration and invasion of adenoid cystic carcinoma cell line ACC-M,and the possible mechanism of PI3K/Akt signaling pathway.METHODS:HBXIP plasmid was transfected into ACC-M.The cells were divided into experimental group (transfected with plasmid pEGFP-N1-HBXIP) control group (non-transfected group) and blank control group (vector group,pEGFP-N1).RT-PCR was used to detect the expression HBXIP in ACC-M;MTT assay,transwell chamber experiments and scratches over the proliferation of HBXIP were utilized individually to evaluate the influence of HBXIP on ACC-M expression,migration and invasion;Western blotting was used to detect the protein expression of Akt,p-Akt,PI3K,p-PI3K and S100A4 after overexpression of HBXIP.Statistical analysis was performed using SPSS 18.0 software package.RESULTS:MTT results showed that the number of surviving cells of experimental group was significantly higher than the control group (P＜0.05);Scratch test results showed that the cell mobility of the experimental group was significantly higher than the control group (P＜0.01);Transwell chamber experiments showed that the number of cell invasion of the experimental group was significantly higher than the control group (P＜0.01);Western blotting results showed that compared with the control group,the expression of p-Akt,p-PI3K and S100A4 in the experimental group with overexpressed HBXIP was relatively increased.CONCLUSIONS:Overexpression of HBXIP gene promotes ACC-M proliferation,invasion and migration.Further,ACC-M proliferation,invasion and migration may be promoted by increased Akt,PI3K phosphorylation and S100A4 protein expression.