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CacyBP/SIP基因siRNA的筛选

Keywords: 乳腺癌,CacyBP/SIP基因,siRNA

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Abstract:

目的探讨靶向钙周期素结合蛋白(CacyBP/SIP)基因的3对小干扰RNAs(smallinterferenceRNA,siRNAs)转染人乳腺癌细胞MDA-MB-231后,能否抑制CacyBP/SIP基因的表达。方法化学合成3对CacyBP/SIP特异性siRNA(CacyBP/SIP-siRNA),分别转染MDA-MB-231细胞,RT-PCR法、Westernblotting方法分别检测转染前后MDA-MB-231细胞的CacyBP/SIPmRNA和蛋白的水平。结果CacyBP/SIP-siRNA001成功转染入人乳腺癌MDA-MB-231细胞,RT-PCR结果显示,siRNA001组CacyBP/SIPmRNA的表达量为NC-siRNA阴性对照组的(25.2±1.34)%,siRNA002和siRNA003则为(64.8±0.58)%和(77.5±1.27)%,CacyBP/SIP-siRNA001转染后能够抑制MDA-MB-231细胞中CacyBP/SIPmRNA,沉默效率为(74.8±0.75)%(P<0.05),Westernblotting证实了siRNA001的沉默作用。结论CacyBP/SIP-siRNA001能显著沉默MDA-MB-231细胞中CacyBP/SIPmRNA和蛋白的表达。

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