2006•Chin J Obstet GynecolRequires access

Enhanced cisplatin cytotoxicity by RNA interfering the excision repair cross-complementing 1 gene in ovarian cancer cell lines

刘国艳, Quanxin Qu, 糜若然, 齐静

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Abstract

目的 探讨RNA干扰技术抑制切除修复交叉互补基因(ERCC)1对卵巢上皮性癌(卵巢癌)细胞顺铂敏感性的影响.方法体外设计、合成针对ERCC1的小分子干扰RNA(siRNA),并转染卵巢癌细胞ES-2,应用RT-PCR、蛋白印迹法(western blot)和免疫组化链霉菌抗生物素蛋白-过氧化物酶(SP)连接法检测转染siRNA后ES-2细胞ERCC1基因和蛋白的表达变化,应用四甲基偶氮唑蓝比色法检测干扰ERCC1后ES-2细胞对顺铂敏感性的变化.结果转染针对ERCC1的siRNA后24、48、72 h,ES-2细胞ERCC1基因和蛋白的表达水平下降;转染ERCC1 siRNA后,ES-2细胞对顺铂的半数抑制量从(10.475±1.713)μg/ml提高到(0.194±0.021)μg/ml,ES-2细胞对顺铂的敏感性增加了53.88倍.结论RNA干扰技术抑制ERCC1能增加卵巢癌细胞ES-2对顺铂的敏感性。

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目的 探讨RNA干扰技术抑制切除修复交叉互补基因(ERCC)1对卵巢上皮性癌(卵巢癌)细胞顺铂敏感性的影响.方法体外设计、合成针对ERCC1的小分子干扰RNA(siRNA),并转染卵巢癌细胞ES-2,应用RT-PCR、蛋白印迹法(western blot)和免疫组化链霉菌抗生物素蛋白-过氧化物酶(SP)连接法检测转染siRNA后ES-2细胞ERCC1基因和蛋白的表达变化,应用四甲基偶氮唑蓝比色法检测干扰ERCC1后ES-2细胞对顺铂敏感性的变化.结果转染针对ERCC1的siRNA后24、48、72 h,ES-2细胞ERCC1基因和蛋白的表达水平下降;转染ERCC1 siRNA后,ES-2细胞对顺铂的半数抑制量从(10.475±1.713)μg/ml提高到(0.194±0.021)μg/ml,ES-2细胞对顺铂的敏感性增加了53.88倍.结论RNA干扰技术抑制ERCC1能增加卵巢癌细胞ES-2对顺铂的敏感性。

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Available abstract

目的 探讨RNA干扰技术抑制切除修复交叉互补基因(ERCC)1对卵巢上皮性癌(卵巢癌)细胞顺铂敏感性的影响.方法体外设计、合成针对ERCC1的小分子干扰RNA(siRNA),并转染卵巢癌细胞ES-2,应用RT-PCR、蛋白印迹法(western blot)和免疫组化链霉菌抗生物素蛋白-过氧化物酶(SP)连接法检测转染siRNA后ES-2细胞ERCC1基因和蛋白的表达变化,应用四甲基偶氮唑蓝比色法检测干扰ERCC1后ES-2细胞对顺铂敏感性的变化.结果转染针对ERCC1的siRNA后24、48、72 h,ES-2细胞ERCC1基因和蛋白的表达水平下降;转染ERCC1 siRNA后,ES-2细胞对顺铂的半数抑制量从(10.475±1.713)μg/ml提高到(0.194±0.021)μg/ml,ES-2细胞对顺铂的敏感性增加了53.88倍.结论RNA干扰技术抑制ERCC1能增加卵巢癌细胞ES-2对顺铂的敏感性。

Key concepts: Cisplatin, Cytotoxicity, Ovarian cancer, Small interfering RNA, Gene, Cancer research, Cell culture, Nucleotide excision repair

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Enhanced cisplatin cytotoxicity by RNA interfering the excision repair cross-complementing 1 gene in ovarian cancer cell lines — Research Paper | ScholarLens