IL-10对稳定转染HBx基因的HL-7702细胞增殖的影响

摘要

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.%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);80 ng/mL的IL-10作用24 h可抑制HL-7702/HBx细胞增殖(P<0.05);流式细胞术显示,80 ng/mL的IL-10作用24 h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CDKN1B mRNA表达量较HL-7702细胞和HL-7702/MOCK细胞降低(P<0.05),80 ng/mL的IL-10作用24 h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CDKN1B mRNA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.