乙肝病毒X蛋白结合蛋白对腺样囊性癌细胞株ACC-2生物学行为的影响

发布时间：2018-10-25 06:16

【摘要】：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。MTT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(P0.05);划痕实验结果显示,实验组细胞迁移率明显低于对照组(P0.01);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(P0.01);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。
[Abstract]:Objective to investigate the effect of hepatitis B virus X protein binding protein (HBXIP) on the biological behavior of adenoid cystic carcinoma cell line ACC-2 and its mechanism. Methods siRNA transfection method was used to inhibit the expression of HBXIP in adenoid cystic carcinoma cell line ACC-2, real-time PCR and Western blotting were used to detect the expression of HBXIP gene and protein, MTT method was used to detect the effect of HBXIP-siRNA on cell proliferation, and transwell method was used to detect the effect of HBXIP-siRNA on cell invasion. The effect of HBXIP-siRNA on cell migration was detected by scratch assay, and the effect of HBXIP-siRNA on Akt,p-Akt,PI3K,p-PI3K and S100A4 protein expression in PI3K/Akt signaling pathway was detected by Western blot. Results HBXIP was highly expressed in ACC-2, and HBXIP-siRNA was successfully transfected into the cell line. The results of MTT showed that the number of cells survived in the experimental group was lower than that in the control group at 48 ~ 72h, the difference was statistically significant (P0.05), and the scratch test showed that the number of cells survived in the experimental group was significantly lower than that in the control group. The cell migration rate in the experimental group was significantly lower than that in the control group (P0.01); transwell chamber experiment results showed that the number of cell invasion in the experimental group was significantly lower than that in the control group (P0.01). In experimental group, the expression of p-AktPI3K and S100A4 decreased with the silencing of HBXIP gene. Conclusion HBXIP can promote the proliferation, migration and invasion of adenoid cystic carcinoma cell line ACC-2, and its mechanism may be related to the promotion of Akt,PI3K phosphorylation and the increase of S100A4 protein expression.
【作者单位】：中国医科大学附属盛京医院口腔颌面外科;辽宁省铁法煤业集团总医院口腔科;沈阳农业大学畜牧兽医学院预防兽医学教研室;
【基金】：辽宁省科学技术计划(2011010215-404)
【分类号】：R739.87

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