环氧合酶-2在HL7702/HBx肝细胞增殖中的作用  

Effect of Cyclooxygenase-2 on the Proliferation of HL7702/HBx Cells

作　　者：李丹[1] 方雪芬 郑碧云[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化科,福州350001

出　　处：《福建医科大学学报》2014年第5期285-289,共5页Journal of Fujian Medical University

基　　金：国家自然基金(81300321);福建省自然基金(2014J01419);福建省临床医学重点专科资助项目{闽卫科教[2012](149号)}

摘　　要：目的探讨环氧合酶-2(COX-2)在稳定表达乙型肝炎病毒X蛋白(HBx)的HL7702肝细胞增殖中的作用及机制。方法 CCK-8法及克隆形成实验检测HL7702/HBx细胞、HL7702/Mock细胞、HL7702细胞增殖情况,RT-PCR法检测上述3种细胞中COX-2 mRNA表达水平,Western-blot法检测COX-2蛋白表达水平,COX-2选择性抑制剂NS-398作用于各组细胞后再检测以上各组细胞增殖情况及COX-2蛋白表达水平的改变。结果细胞增殖实验及克隆形成实验提示,HL7702/HBx组细胞增殖能力强于HL7702/Mock和HL7702组(P<0.05),克隆形成率也更高(P<0.05)。RT-PCR检测结果提示,相对于HL7702/Mock细胞和HL7702组细胞,HL7702/HBx组细胞内COX-2的mRNA相对表达量明显增高(P<0.05)。Western-blot检测提示,HL7702/HBx组细胞中的COX-2蛋白相对表达水平较高(P<0.05),而HL7702/Mock及HL7702组细胞之间则无明显差别。NS-398以时间依赖的方式部分抑制各组细胞的增殖能力,对HL7702/HBx组细胞增殖的抑制强于其他2组细胞(P<0.05)。经50μmol/L NS-398处理后,3组细胞的COX-2蛋白水平均显著降低,此现象在HL7702/HBx组细胞中更加明显(P<0.05)。结论 HBx蛋白可以上调肝细胞COX-2表达,促进肝细胞增殖,选择性COX-2抑制剂可部分逆转该作用。Objective To investigate the effect of cyclooxygenase‐2（COX‐2） on the cell prolifera‐tion in HL7702 cells w hich express hepatitis B virus X protein （HBx ） stably . Methods Cell‐counting Kit‐8 （CCK‐8） assay and clone formation assay were explored to detect the proliferation of HL 7702/HBx , HL7702/Mock and HL7702 cells ,RT‐PCR and Western‐bolt were used to examine the level of COX‐2 mRNA and protein expression in cells of three groups . The level of cell proliferation and COX‐2 protein expression were measured respectively after treated with selective COX‐2 inhibitor NS‐398 . Results CCK‐8 assay and plate colony formation assay displayed the proliferation rate of HL 7702/HBx was higher than that of HL7702/Mock and HL7702 cells . The level of COX‐2 mRNA and protein expression was greater in HL7702/HBx cells compared with that of HL7702/Mock and HL7702 cells（P〈0 .05） . COX‐2 selective inhibitor NS‐398 suppressed growth of three groups of cells in a time‐dependent manner . The proliferation inhibition rate of HL7702/HBx cells was significantly higher than that of other two groups in three time points respectively （P〈0 .05） . After treated with 50 μmol/L NS‐398 for 72 h ,COX‐2 expres‐sion was suppressed in all group , however , this phenomenon is more obvious in HL7702/HBx cells （P〈0 .05） . Conclusion Regulate the expression of COX‐2 is one of the pathway for the effect of HBx on the HL7702 proliferation ,COX‐2 inhibitor can suppress the cell proliferation for the greater part in‐duced by this pathway .

关 键 词：病毒蛋白质类 肝炎病毒 乙型 细胞增殖 前列腺素内过氧化物合酶类 同工酶类 

分 类 号：R512.62[医药卫生—内科学]