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

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

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。

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 （transfeeted 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; MTF 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： MTF 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 groupwith 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.