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本研究主要考察siRNA对于逆转P-糖蛋白介导的肿瘤多药耐药的作用。以MDR1基因为靶标,在线设计MDR1基因的siRNA序列并利用生物学软件进行优化,经化学合成后转染入体外培养的乳腺癌耐药细胞系MCF-7/ADR中,经实时定量荧光PCR和Western blot定量分析MDR1基因的表达,以MTT法检测转染细胞对多柔比星的敏感性。最终设计并合成了8条MDR1基因siRNA序列,其中4条能有效抑制MDR1基因mRNA的表达。siRNA1沉默效率最高,可特异性地沉默MDR1基因,有效逆转乳腺癌细胞的多药耐药性。
This study mainly investigated the effect of siRNA on reversing P-glycoprotein-mediated multidrug resistance in tumors. The MDR1 gene was designed as a target to design the siRNA sequence of MDR1 gene online and optimized by biological software. The gene was transfected into the MCF-7 / ADR cell line which was cultured in vitro. After real-time quantitative PCR and real- Western blot quantitative analysis of MDR1 gene expression MTT assay transfected cells to doxorubicin sensitivity. Finally, 8 MDR1 gene siRNA sequences were designed and synthesized, of which 4 were effective in inhibiting the expression of MDR1 gene mRNA. siRNA1 has the highest silencing efficiency and can specifically silence the MDR1 gene, effectively reversing the multi-drug resistance of breast cancer cells.