IL-10-HGF融合基因对肝星状细胞LX-2增殖的抑制作用

The Proliferation of Hepatic Stellate Cell LX-2 Inhibited by Human Interleukin-10 and Human Hepatocyte Growth Factor Fused Gene

目的探讨人白介素-10(IL-10)与人肝细胞生长因子(HGF)融合基因对肝星状细胞LX-2增殖的抑制作用。方法采用PCR方法,分别构建pcDNA3.1-flag-IL-10、pcDNA3.1-flag-HGF与pcDNA3.1-flag-IL-10-HGF真核表达载体,瞬时转染肝星状细胞LX-2;利用RT-PCR和Western blot检测LX-2细胞中IL-10、HGF的表达;选用MTT法检测转染24、48和72 h后LX-2细胞的增殖情况。结果成功构建了IL-10-HGF融合基因真核表达载体pcDNA3.1-flag-IL-10-HGF。与pcDNA3.1-flag-IL-10和pcDNA3.1-flag-HGF组比较,pcDNA3.1-flag-IL-10-HGF融合组48 h和72 h LX-2细胞数减少(n=3,P<0.05)。结论 IL-10-HGF融合基因可有效抑制肝星状细胞LX-2的增殖。

Objective The study investigated the action of human interleukin-10 and human hepatocyte growth factor fused gene for the proliferation of cells LX-2.Methods The eukaryotic expression vector of pcDNA3.1-flag-IL-10,pcDNA3.1-flag-HGF and pcDNA3.1-flag-IL-10-HGF were constructed by PCR.They were transiently transfected with hepatic stellate cell LX-2.Then,the expression of human interleukin-10 and human hepatocyte growth factor were detected by RT-PCR and Western blot.The proliferation of LX-2 cells was checked by MTT method after transfected 24h,48h and 72h.Results The eukaryotic expression vector pcDNA3.1-flag-IL-10-HGF including human interleukin-10 and human hepatocyte growth factor fused gene was successfully constructed.Compared with the groups of IL-10 and HGF,the LX-2 cell number were decreased in IL-10-HGF fused gene group after transfected 48h and 72h（n=3,P0.05）.Conclusion The IL-10-HGF fused gene can suppress the proliferation of hepatic stellate cell LX-2.