水通道蛋白9表达水平影响HepG2肝癌细胞的生物学行为及对As_2O_3的敏感性  

The expression of AQP9 in Hep G2 cells affects cell biological behaviors and sensitivity to As_2O_3

作　　者：唐洁[1] 王川[1] 姜政[1] 

机构地区：[1]重庆医科大学附属第一医院消化内科,重庆400016

出　　处：《细胞与分子免疫学杂志》2015年第6期769-774,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81070318);重庆市卫生局基金(2010-2-100)

摘　　要：目的研究水通道蛋白9(AQP9)表达水平对三氧化二砷(As2O3)诱导Hep G2肝癌细胞凋亡及对其生物学行为的影响。方法采用MTT法筛选As2O3对肝癌细胞Hep G2增殖抑制作用;将重组质粒p EGFP-N1-AQP9和pshRNA-AQP9转染肝癌细胞,反转录PCR检测AQP9 mRNA的表达,Western blot法检测AQP9蛋白表达;将As2O3加入转染后及未处理的Hep G2细胞,MTT法检测细胞增殖抑制率;流式细胞术分析周期和凋亡情况;TranswellTM实验验证迁移侵袭能力的改变;酶标仪检测caspase-3活性。结果重组质粒对AQP9有明显的过表达及抑制表达作用,转染p EGFP-N1-AQP9组肝癌细胞的增殖抑制作用明显大于单纯As2O3处理组,侵袭及迁移能力显著减弱,细胞周期停滞在G0/G1期,凋亡明显多于单纯As2O3处理组,活化的caspase-3活性最高。转染pshRNA-AQP9组细胞的增殖抑制作用及caspase-3活性小于单纯As2O3处理组,侵袭迁移能力强于单纯As2O3处理组,细胞周期停滞在G0/G1期的数量及凋亡率较少。结论调节AQP9的表达影响Hep G2肝癌细胞的生物学行为并影响As2O3对Hep G2肝癌细胞的敏感性。Objective To investigate the effect of aquaporin 9 （AQP9） expression level on the apoptosis and biological behaviors of HepG2 cells induced by arsenic trioxide （As2O3）. Me,otis The effects of different concentration As2O3 on the cell proliferation were measured by MTT assay, and IC50 was calculated. The recombinant plasmids pEGFP-N]-AQP9 and pshRNA-AQP9 were transfected into HepG2 cells. The expression of AQP9 mRNA and protein were detected by reverse transcription PCR and Western blotting, respectively. Then As2O3 was added to plasmid-transfected cells and untreated HepG2 cells. Cell proliferation was detected by MTT assay, cell cycle and apoptosis rate were determined by flow cytometry, and ceil invasion and migration ability were examined by TrenswellTM （ Boyden Chamber） assay. The activity of caspase-3 was detected by microplate reader. Results The recombinant plasmids remarkably influenced the expression of AQPg. Compared with the group treated with As2O3 alone （control group）, the cell proliferation, invasion and migration of HepG2 cells transfected with pEGFP-N]-AQP9 were attenuated significantly, while the cells transfected with pshRNA-AQP9 increased. The increasing population of apoptotic cells, augmented caspase 3 activity and highest percentage of cells stagnating at G0/GI phase were observed in HepG2 cells transfected with pEGFP-N1-AQP9 as compared with the control HepG2 cells; in contrast, the HepG2 cells transfected with pshRNA-AQP9 presented with lower caspase 3 activity and stronger invasion and migration ability than the control HepG2 cells did. Conclusion The alterations in the expression of AQP9 could affect the biological behaviors of HepG2 cells, but also their sensitivity to As2O3.

关 键 词：AQP9 三氧化二砷 肝癌 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤] R392.12[医药卫生—临床医学]