IL-10对稳定转染HBx基因的HL-7702细胞增殖的影响  

The Effect of IL-10 on the Proliferation of HL-7702 Cells Stably Expressing HBx Gene

作　　者：张洁[1] 郑碧云[1] 陈治新[1] 李丹[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》2017年第2期77-81,共5页Journal of Fujian Medical University

基　　金：福建省临床医学重点专科资助项目(闽卫科教[2012]149号);福建省自然科学基金青年项目(2016J05189);福建省卫生和计划生育委员会中青年骨干重点项目(2014-ZQN-ZD-9)

摘　　要：目的探讨白细胞介素-10(IL-10)在稳定转染乙型肝炎病毒X(HBx)基因的HL-7702肝细胞增殖的作用及机制。方法 CCK-8法测定HBx基因对HL-7702细胞增殖的作用及IL-10对HL-7702/HBx细胞增殖的作用;流式细胞术测定IL-10对HL-7702/HBx细胞凋亡的影响;RT-PCR法测定HBx基因对HL-7702细胞表达CDKN1B蛋白质mRNA的作用及IL-10对HL-7702/HBx表达CDKN1B mRNA的作用。结果 CCK-8法显示,HL-7702/HBx细胞比HL-7702细胞和HL-7702/MOCK细胞增殖速度明显加快(P<0.05);80ng/mL的IL-10作用24h可抑制HL-7702/HBx细胞增殖(P<0.05);流式细胞术显示,80ng/mL的IL-10作用24h对HL-7702细胞、HL-7702/HBx细胞及HL7702/MOCK细胞凋亡均无影响;RT-PCR法显示,HL-7702/HBx细胞CDKN1BmRNA表达量较HL-7702细胞和HL-7702/MOCK细胞降低(P<0.05),80ng/mL的IL-10作用24h后,HL-7702/HBx细胞较HL-7702/HBx空白组CDKN1B mRNA表达量上调(P<0.05)。结论 HBx基因可促进HL-7702细胞增殖,IL-10可抑制HL-7702/HBx细胞增殖而对其凋亡无影响。HBx基因可能通过下调HL-7702/HBx细胞CDKN1BmRNA的表达量而促进细胞增殖,IL-10可能通过上调HL-7702/HBx细胞CDKN1B mRNA的表达量而抑制细胞增殖。Objective To investigate the effect of interleukin-10（IL-10） on the proliferation of HL-7702 cells stably expressing hepatitis B virus X protein（HBx） gene.Methods Cell-counting Kit-8（CCK-8） assay were explored to detect the effect of HBx gene on the proliferation of HL-7702 cells and the effect of IL-10 on the proliferation of HL-7702 cells stably expressing HBx gene.Flow cytometry was used to analyze the effect of IL-10 on the apoptosis of HL-7702/HBx cells.RT-PCR was used to detect the effect of HBx gene on the expression of CDKN1B mRNA in HL-7702 cells and the effect of IL-10 on the expression of CDKN1B mRNA in HL-7702/HBx cells.Result The data of CCK-8 assay showed that the proliferation rate of HL-7702/HBx cells was significantly faster than HL-7702 and HL-7702/MOCK cells（P〈0.05）.The proliferation of the HL-7702/HBx cells was significantly inhibited after being treated with 80 ng/mL IL-10 for 24 hours（P〈0.05）.In addition,treatment of IL-10 of 80 ng/mL for 24 hours had no effects on the apoptosis of HL-7702,HL-7702/MOCK and HL-7702/HBx cells.Compared with HL-7702 and HL-7702/MOCK cells,the expression of CDKN1B mRNA level in HL-7702/HBx cells was decreased（P〈0.05）.After the treatment of 80 ng/mL IL-10 for 24 hours,the expression of CDKN1B mRNA level in HL-7702/HBx cells had increased（P〈0.05）.Conclusions HBx gene can promote the proliferation of HL-7702 cells.IL-10 can inhibit the proliferation of HL-7702/HBx cells but has no effect on the apoptosis.HBx gene probably enhances the proliferation of HL-7702/HBx cells by down-regulating the expression of CDKN1B mRNA.IL-10 can inhibit the proliferation of HL-7702/HBx cells which was probably related to the up-regulation of the expression of CDKN1B mRNA.

关 键 词：病毒蛋白质类 肝细胞 肝炎病毒 乙型 白细胞介素10 ＊细胞转化 肿瘤 细胞增殖 

分 类 号：R512.62[医药卫生—内科学]