XPD对肝癌HepG2增殖及REDD1基因表达的影响-医学专业论文.docxVIP

摘要 摘要 万方数据 万方数据 摘 要 目的：探讨人着色干皮病基因 D(xeroderma pigmentosum D，XPD)对人肝癌 细胞 HepG2 增殖凋亡及 REDD1 基因表达的影响。 方法：实验将重组质粒 pEGFP-N2/XPD 及空载质粒 pEGFP-N2 通过脂质体 （Lipofectamine?2000）介导的瞬时转染方式转入人肝癌细胞HepG2中。实验 分为 4 组，分别为空白组、脂质体组、pEGFP-N2 组、pEGFP-N2/XPD 组。以 基因 绿色荧光 蛋白在荧光 显微镜表 达观察 转染情况 ；逆 转录聚合 酶链反应 (RT-PCR)、蛋白印迹（Western blot）法测细胞中 XPD、REDD1 基因的 mRNA、 蛋白质的表达水平；流式细胞仪测细胞凋亡变化；MTT 法测细胞增殖。 结 果： 荧 光显 微镜 观察 到转 染了 重 组质 粒 pEGFP-N2/XPD 及 空载 质粒 pEGFP-N2 的细胞绿色荧光，表明转染成功。RT-PCR 及 Western blot 结果显示： 与其他 3 组相比较，XPD 组中 XPD、REDD1 的 mRNA 及蛋白表达量明显升高 （均 P 0.01）。MTT 法结果示 XPD 组较其他 3 组细胞增殖活性显著降低（均 P 0.01）。流式细胞仪结果示 XPD 组细胞凋亡率明显升高达到 45%（均 P 0.01）。 结论：XPD 能上调抑癌基因 REDD1 的表达，并能显著抑制肝癌细胞 HepG2 的增殖促进其凋亡。 关键词：肝肿瘤；XPD 基因；REDD1 基因 II Ab Abstract ABSTRACT Objective：To investigate the effects of transfected XPD gene on the growth of hepatoma cell HepG2 and the expression of REDD1 gene in the cells in vitro. Methods: The recombinant plasmid pEGFP-N2/XPD and experimental plasmid pEGFP-N2 through empty liposomes (Lipofectamine ? 2000) mediated transient transfection into human hepatoma cell line HepG2 . Experiment was divided into four groups, respectively, human hepatoma cell line HepG2( blank control group),liposome transfection human hepatoma cell line HepG2 group (Lip group),empty plasmid transfection human hepatoma cell line HepG2 - pEGFP - N2 group (N2 group),recombinant plasmid transfection human hepatoma cell line HepG2 - pEGFP - N2 - XPD group (XPD group). Fluorescence microscopy of green fluorescent protein gene expression, reverse transcription polymerase chain reaction (RT-PCR), Western blotting (Western blot) measured in each group XPD, REDD1 gene mRNA,and protein expression levels. The cell was apoptosis measured by flow cytometry; The cell proliferation was detected by MTT. Results: Fluorescence microscopy to transfected with the recombinant plasmid pEGFP-N2/XPD and empty plasmid pEGFP-N2 cells green fluorescence, indicating successful transfection. RT-PCR and Western blot showed that compared w