论文部分内容阅读
目的探讨针对多药耐药基因(MDR-1)的小分子干扰RNA(siRNAs)和反义脱氧寡核苷酸(asODNs)联合应用逆转人乳腺癌细胞MCF-7/ADR的作用效果。方法设计并合成针对MDR-1基因同一序列的siRNAs和asODNs及阴性对照siRNAs,采用转染试剂lipofectamineTM2000分别转染人乳腺癌耐药细胞MCF-7/ADR;利用RT-PCR检测MDR-1mRNA和Western blot检测MDR-1蛋白质的表达;采用罗丹明123外排实验检测P-gp的转运功能,MTT法检测MCF-7/ADR细胞对阿霉素的耐药逆转效果。结果siRNAs、asODNs、asODNs和siRNAs联合应用均能降低MDR-1mRNA及其蛋白质表达,提高P-gp的转运功能,使细胞对阿霉素的敏感性明显恢复;asODNs和siRNAs联合应用效果明显提高;低浓度siRNAs(200nmol/L)比高浓度asODNs(5μmol/L)的效果强。结论siRNAs、asODNs能有效逆转人乳腺癌细胞MCF-7/ADR的多药耐药,asODNs和siRNAs联合应用效果明显加强。
Objective To investigate the effect of multidrug resistance gene (MDR-1) combined with small interfering RNA (siRNAs) and antisense oligodeoxynucleotides (ODNs) on reversing human breast cancer cell line MCF-7 / ADR. Methods siRNAs and asODNs against the same sequence of MDR-1 gene and negative control siRNAs were designed and synthesized. The human breast cancer cell line MCF-7 / ADR was transfected with the lipofectamineTM2000 transfection reagent. The mRNA and protein expressions of MDR-1 mRNA and Western blot was used to detect the expression of MDR-1 protein. The rhodamine 123 efflux assay was used to detect the translocation of P-gp. MTT assay was used to detect the reversal effect of doxorubicin on MCF-7 / ADR cells. Results The combination of siRNAs, asODNs, asODNs and siRNAs could decrease the expression of MDR-1 mRNA and protein, enhance the transport function of P-gp and restore the sensitivity of cells to doxorubicin. The combined effect of asODNs and siRNAs was significantly improved. Low concentrations of siRNAs (200nmol / L) than the high concentration of asODNs (5μmol / L) effect. Conclusion siRNAs and asODNs can effectively reverse the multidrug resistance of human breast cancer cells MCF-7 / ADR. The combined effect of asODNs and siRNAs was significantly enhanced.