摘要
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(small interference RNA,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、Western blotting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/SIP mRNA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),Western blotting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/SIP mRNA和蛋白的表达。
Objective To explore the effects of silencing CacyBP /SIP expression by small interference RNA( siRNA) of human breast cancer MDA-MB-231 cells. Methods Chemically synthesized siRNAs targeting CacyBP / SIP were transfected into MDA-MB-231 cells. The expression of CacyBP / SIPmRNA and protein was detected by real-time quantitative polymerase chain reaction( PCR) and Western blotting( P < 0. 05).Results CacyBP /SIP-siRNA was successfully transfected into MDAMDA-MB-231 cells. The expression of CacyBP / SIP mRNA was much lower in the CacyBP / SIP-siRNA001 transfected group as compared with negative-control group. The expression of CacyBP / SIP mRNA was( 25. 2 ± 1. 34) %,( 64. 8 ± 0. 58) %,( 77. 5 ±1. 27) % in the siRNA001,siRNA002 and 003 transfected group as compared with negative-control group,respectively. and its mRNA level were reduced by about( 74. 8 ± 0. 75) % in CacyBP / SIP-siRNA001 expressing cells( P < 0. 05). RT-PCR and Western blot showed that mRNA and protein level were markedly decreased. Conclusion CacyBP / SIP-siRNA001 can silence CacyBP / SIP mRNA and protein expression in MDA-MB-231 cells.
出处
《宁夏医科大学学报》
2013年第8期853-855,859,共4页
Journal of Ningxia Medical University
基金
宁夏自然科学基金重点项目(NZ10113)
