摘要
目的:探讨激活素受体相互作用蛋白2(ARIP2)在肝细胞内的表达形式及其介导的生物学作用.方法:采用免疫组织化学及细胞化学染色、Western blot检测ARIP2蛋白在小鼠肝组织及肝癌细胞系Hepal-6细胞内的表达;采用激活素特异应答的CAGA-lux报告基因质粒分析ARIP2对激活素诱导的特异基因转录的影响;MTT法检测ARIP2过表达对Hepal-6细胞增殖的彰响.结果:ARIP2蛋白可在小鼠肝组织及Hepal-6细胞表达.激活素A刺激Hepal-6细胞ARIP2蛋白表达呈时间依赖性升高,于24 h达到高峰;12 h、24 h测定值与对照组相比有显著性意义(1.01±0.16,1.62±0.26 vs 0.82±0.11.P<0.05).pcDNA3-ARIP2转染Hepal-6细胞.可以明显抑制激活素诱导的特异基因转录;MTT检测显示激活素A(5μg/L、10μg/L)可以明显抑制Hepal-6细胞增殖,其570 nm吸光度值与对照组相比有显著差异(1.59±0.03、1.49±0.04 vs 1.79±0.07,P<0.05,P<0.01).在Hepal-6细胞内过表达ARIP2可以阻断激活素A对Hepal-6细胞的增殖抑制作用.结论:ARIP2有望成为治疗激活素诱导的肝脏损伤性疾病的基因调控靶点.
AIM： To investigate the activin receptorinteracting protein 2 （ARIP2） expression and its biological function in hepatocytes. METHODS： Expression of ARIP2 in mouse liver tissue and hepatoma cell line Hepal-6 cells was detected by Western blot, immunohistochemistry and cytochemical staining. Effect of ARIP2 on activin-induced gene transcription was analyzed using CAGA-lux plasmid. Effect of overexpression of ARIP2 on the proliferation of Hepal-6 cells was assayed with MTT method.
RESULTS： ARIP2 was expressed in mouse liver tissue and Hepal-6 cells. The expression of ARIP2 in activin A-stimulated Hepal-6 cells was increased in a time-dependent manner, and peaked at 24 h. There was a significant difference in the expression level of ARIP2 on Hepal-6 cells at 12 and 24 h in contrast with the control group （1.01 ± 0.16, 1.62 ± 0.26 vs 0.82 ± 0.11, P 〈 0.05, P 〈 0.01）. pcDNA3-ARIP2-transfected Hepal-6 cells obviously suppressed the gene transcription induced by activin A. MTT assay displayed that activin A （5 μg/L and 10 μg/L） remarkably inhibited the proliferation of Hepal-6 cells, the A570 nm value was 1.59 ± 0.03 and 1.49 ± 0.04 vs 1.79±0.07, respectively （P 〈 0.05, P 〈 0.01）. ARIP2 over-expression in Hepal-6 cells significantly blocked the inhibitory effects of activin A （5 μg/L and 10 μg/L） on the proliferation of Hepal-6 cells.
CONCLUSION： ARIP2 can be expected to become a regulation target of genes in treatment of liver injury induced by activin.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第4期350-355,共6页
World Chinese Journal of Digestology
基金
国家自然科学基金
No.30170478和No.30771957
吉林省科技厅资助项目
No.20060928-01~~
