RNA干扰乳腺癌耐药蛋白基因表达改变乳腺癌耐药细胞耐药性的研究  

Changes in drug resistance of breast cancer MCF -7/ADR cell line by inhibiting breast cancer re- sistance protein gene expression using RNA interference

作　　者：杨永刚[1] 周敏[1] 江学庆[1] 

机构地区：[1]武汉市中心医院甲状腺乳腺外科,430014

出　　处：《中华实验外科杂志》2016年第4期933-935,共3页Chinese Journal of Experimental Surgery

摘　　要：目的利用RNA干扰技术抑制乳腺癌耐药细胞株MCF-7／ADR乳腺癌耐药蛋白（BCRP）基因的表达并提高对阿霉素（ADR）的敏感性。方法针对BCRP的mRNA序列，设计两条RNA干扰链（BCRP-A和BCRP-B），构建出小干扰RNA（siRNA）表达载体pG—BCRP—AsiRNA-增强型绿色荧光蛋白（EGFP）和pG—BCRP-BsiRNA—EGFP。实验分为4组：A、B、C3组分别转染BCRP-A、BCRP-B、HK-siRNA（对照组）；D组转染过程中未加入任何质粒（空白组）。通过反转录-聚合酶链反应（RT-PCR）和Westernblot检测siRNA阻断BCRP基因表达的效率，应用噻唑蓝（MTT）法分析siRNA干扰后MCF-7／ADR的耐药性变化。结果RT—PCR和Westernblot法检测结果表明BCRP—A和BCRP-B两条干扰链干扰效率不一，前者较后者对BCRPmRNA和蛋白质表达抑制率均明显优于后（P〈0．05）；MTr法分析结果说明BCRP-A组对ADR的药物敏感性明显提高，其IC50为（184．6±25．3）ng／ml，与BCRP—B干扰组和对照组比较差异有统计学意义（P〈0．05）。结论RNA干扰技术可用于抑制BCRP基因的表达，并提高对ADR的敏感性，但不同干扰链的干扰效率差异较大。Objective To inhibit the expression of breast cancer resistance protein （BCRP） gene in breast cancer MCF -7/ADR cell line and to improve the sensitivity to adriamycin. Methods Two small interfering RNAs （ siRNAs ： BCRP - A and BCRP - B） were designed according to the mRNA sequence of BCRP, and the siRNA expression vectors pG - BCRP - A siRNA - enhanced green fluorescent protein （EGFP） and pG - BCRP - B siRNA - EGFP were constructed. Four groups were set up. The MCF - 7/ ADR cells in groups A, B, and C were treated by pG - BCRP - A siRNA - EGFP, pG - BCRP - B siRNA - EGFP and HK - siRNA, respectively. The cells in group D were untreated. Reverse transcriptase - polymer- ase chain reaction （ RT - PCR） and Western blotting were used to verify the efficiency of siRNA in inhibi- ting the expression of BCRP in MCF-7/ADR. Methyl thiazol tetrazolium （MTT） assay was used for ana- lysing the change of adriamycin resistance. Results The RT - PCR and Western blotting assay showed that the interference efficiency of the two interference chains was different, and BCRP - A was significantly better than BCRP- B （P〈0.05）. The results of MTr showed that the ICso of BCRP -A group was （184.6 ± 25. 3 ） ng/ml, and the drug sensitivity of BCRP - A group was significantly higher than the other two groups （P 〈 0.05）. Conclusion RNA interference technology can be used to inhibit the expression of BCRP gene, and improve the sensitivity to adriamycin, but the interference efficiency of different interference chains is different.

关 键 词：RNA干扰 乳腺癌 乳腺癌耐药蛋白基因 阿霉素耐药性 

分 类 号：R512.62[医药卫生—内科学]