UHRF1基因在人卵巢癌细胞株OVCAR-3中的表达及意义  

Expression of UHRF1 gene in ovarian cancer cell line OVCAR-3 and its significance

作　　者：周碧芳[1] 邓斐[1] 周学惠[1] 吴强[1] 

机构地区：[1]江苏省肿瘤医院妇瘤科,南京210009

出　　处：《现代妇产科进展》2012年第10期748-751,共4页Progress in Obstetrics and Gynecology

摘　　要：目的:探讨针对UHRF1基因的小干扰RNA(siRNA)转染人卵巢癌细胞株OVCAR-3后UHRF1表达的变化及细胞的增殖和凋亡,为UHRF1基因作为新的靶点治疗卵巢癌提供依据。方法:用siRNA基因沉默技术,将体外化学合成的针对UHRF1基因的siRNA转染至OVCAR-3细胞(转染组),并以转染阴性对照siRNA序列的细胞作为阴性对照组,未经转染的细胞作为空白组;实时荧光定量PCR和Western blot分别检测转染前后细胞UHRF1 mRNA和蛋白表达水平的变化;CCK-8实验和流式细胞术分别检测沉默UHRF1基因后对细胞增殖和凋亡的影响。结果:实时荧光定量PCR结果显示,siRNA转染后UHRF1基因在卵巢癌细胞株OVCAR-3中表达显著降低(P<0.01)。Western blot结果显示,转染组、阴性对照组和空白组细胞UHRF1蛋白相对表达量分别为0.16±0.04、0.33±0.03、0.35±0.07;转染组细胞UHRF1蛋白表达水平显著低于阴性对照组和空白组(P<0.05),而阴性对照组和空白组之间无显著差异(P>0.05)。CCK-8实验结果显示,转染后48,72,96h,转染组细胞增殖率均显著低于空白组(P<0.01)。流式细胞术结果显示,转染组、阴性对照组细胞凋亡率分别为(15.83±2.22)%、(7.46±2.93)%,差异显著(P<0.01)。结论:UHRF1基因沉默后可显著抑制卵巢癌OVCAR-3细胞UHRF1的表达,并抑制细胞增殖,促进细胞凋亡。Objective：To investigate the expression of UHRF1,the changes in cell proliferation and apoptosis after small interfering RNA（siRNA） for UHRF1 gene were transfected into the human ovarian cancer cell line OVCAR-3,and to provide the basis for establishing UHRF1 gene as a new target for the treatment of ovarian cancer.Methods：Using the siRNA gene silencing technique,chemical synthesis small interfering RNA establishing for UHRF1 gene sequence were transfected into the human ovarian cancer cell line OVCAR-3（transfection group）.At the same time the negative control siRNA sequence was also transfected into the OVCAR-3 cell as the negative control group.Non-transfected cell was considered as the blank group.Real-time quantitative PCR and Western blot assay separately were used to compare the mRNA and protein level of UHRF1 before and after siRNA transfection.CCK-8 assay and flow cytometry were used to investigate the cell proliferation and cell apoptosis after UHRF1 gene silencing.Results：Real-time quantitative PCR results showed the expression of UHRF1 was significantly decreased in OVCAR-3 cells after the transfection of siRNA（P0.01）.The Western blot results showed the relative expression between the UHRF1 protein and the β-actin protein in transfection group,negative control group and blank group respectively were 0.16±0.04,0.33±0.03,0.35±0.07.The UHRF1 protein level in transfection group was lower than negative control group and blank group,the difference was statistically significant（P0.05）.However,the comparison between the negative control group and the blank group was not statistically significant（P0.05）.The experiment of CCK-8 and the flow cytometric analysis showed that UHRF1 gene silencing can effectively inhibit OVCAR-3 cell proliferation.48,72,96 hours after transfection,the proliferation rates of transfected cells were lower than those of the negative control group and blank group,the difference was statistically significant（P0.01）.At the same time,inhibition of UHRF1 gene

关 键 词：UHRF1基因 卵巢肿瘤 OVCAR-3细胞株 RNA干扰 细胞增殖 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]