| EZH2基因在人卵巢癌顺铂耐药株中的表达及其对细胞耐药性的影响 |
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| 引用本文： | 胡沙,王静,李智敏,郭剑锋,于利利,杨纯,王泽华.EZH2基因在人卵巢癌顺铂耐药株中的表达及其对细胞耐药性的影响[J].肿瘤,2010,30(9). |
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| 作者姓名： | 胡沙 王静 李智敏 郭剑锋 于利利 杨纯 王泽华 |
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| 作者单位： | 华中科技大学同济医学院附属协和医院妇产科,武汉,430022 |
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| 基金项目： | 国家自然科学基金资助项目 |
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| 摘 要： | 目的:检测果蝇zeste基因增强子同源物2(enhancer of zeste homolog 2, EZH2)基因在人卵巢癌顺铂(cisplatin, DDP)耐药细胞株A2780/DDP及其亲本细胞株A2780中的表达差异,探讨以EZH2基因为靶向的RNA干扰能否有效逆转A2780/DDP细胞对DDP的耐药性.方法:采用实时荧光定量PCR(real-time fluorogenic quantitative PCR,RFQ-PCR)和Western印迹法检测EZH2 mRNA和蛋白在A2780/DDP及A2780细胞中的表达差异.人工构建4个靶向沉默EZH2基因的短发夹状RNA(short hairpin RNA,shRNA)质粒表达载体,脂质体法瞬时转染至A2780/DDP细胞,RFQ-PCR检测EZH2基因的沉默水平并挑选沉默效果最佳的质粒载体,G418加压筛选得到转染稳定的细胞株.RFQ-PCR和Western印迹法检测稳定转染后A2780/DDP细胞中EZH2基因的表达,MTT法检测转染细胞对DDP的耐药性.结果:A2780/DDP细胞中EZH2 mRNA和蛋白的表达量分别是A2780细胞表达量的(3.50±1.06)和(2.10±0.29)倍,差异有统计学意义(P<0.01).稳定转染EZH2 shRNA后的A2780/DDP细胞,EZH2的mRNA及蛋白水平的表达量分别下降(83.66±5.65)%和(77.57±3.90)%(P<0.001),其对DDP的半数抑制浓度(half inhibitory concentration,IC50)值下降(61.33±11.40)%,差异有统计学意义(P<0.05). 结论:EZH2基因在A2780/DDP细胞中的表达明显升高,沉默EZH2基因能有效逆转A2780/DDP细胞对DDP的耐药性.
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| 关 键 词： | 卵巢肿瘤 RNA 小分子干扰 抗药性 肿瘤 EZH2基因 |
Expression of EZH2 gene in cisplatin-resistant ovarian cell line A2780/DDP and its effect on cisplatin resistance |
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| HU Sha,WANG Jing,LI Zhi-min,GUO Jian-feng,YU Li-li,YANG Chun,WANG Ze-hua.Expression of EZH2 gene in cisplatin-resistant ovarian cell line A2780/DDP and its effect on cisplatin resistance[J].Tumor,2010,30(9). |
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| Authors: | HU Sha WANG Jing LI Zhi-min GUO Jian-feng YU Li-li YANG Chun WANG Ze-hua |
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| Institution: | HU Sha,WANG Jing,LI Zhi-min,GUO Jian-feng,YU Li-li,YANG Chun,WANG Ze-hua(Department of Obstetrics and Gynecology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China) |
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| Abstract: | Objective:To examine the enhancer of zeste homolog 2(EZH2) gene expression in cisplatin-resistant ovarian cancer cell line A2780/DDP and its parental cell line A2780,and to investigate whether silencing EZH2 by RNA interference could reverse cisplatin resistance of A2780/DPP cells.Methods:Expressions of EZH2 mRNA and protein in cells were detected by real-time fluorogenic quantitative PCR(RFQ-PCR) and Western blotting,respectively.The difference of EZH2 expression was compared between A2780/DDP and A2780 ce... |
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| Keywords: | Ovarian neoplasms RNA small interfering Drug resistance neoplasms EZH2 gene |
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