【摘要】 目的探讨N乙酰氨基葡萄糖转移酶Ⅴ(GnTⅤ)对细胞基因表达和功能的影响。方法用WesternBlot验证GnTⅤ的反义cDNA稳转的细胞株AsGnTⅤ7721(AsGnTⅤSMMC7721)中GnTⅤ表达,用基因芯片技术比较AsGnTⅤ7721与7721(SMMC7721)细胞的基因表达差异,同时用RTPCR和WesternBlot检测AsGnTⅤ7721细胞中未折叠蛋白质反应信号通路关键分子Bip和XBP1的表达变化。结果基因芯片检测到的AsGnTⅤ7721细胞中的基因表达变化表明,细胞中产生未折叠蛋白质反应。同时AsGnTⅤ7721细胞中未折叠蛋白质反应信号途径中的关键分子Bip的mRNA和蛋白表达升高,XBP1mRNA被剪接。结论GnTⅤ表达受阻能导致细胞产生未折叠蛋白质反应。更多还原

【Abstract】 Purpose To study the effects of N-acetylglucosaminyltransferase Ⅴ on gene expression and function of cells. Methods cDNA microarray analysis was used to study the difference in gene expression profile between 7721(SMMC-7721) and AsGnT-Ⅴ7721(AsGnT-Ⅴ SMMC-7721) cells.The mRNA and protein level of Bip in 7721,AsGnT-Ⅴ7721 were detected by RT-PCR and Western Blot.XPB1 mRNA splicing was confirmed by RT-PCR. Results cDNA microarray analysis demonstrated that there exists unfolded protein response in AsGnT-V7721 cells.Bip mRNA and protein expression was elevated and XBP1 mRNA was spliced in AsGnT-Ⅴ7721. Conclusions Suppression of N-acetylglucosaminyltransferaseV expression induces unfol- ded protein response in hepatocellular carcinoma cell line 7721.更多还原