乳腺癌耐药蛋白重组腺病毒干扰载体构建及其干扰作用的观察

Construction of RNAi recombinated adenovirus targeting BCRP and its inhibitive effects on BCRP

目的:观察腺病毒加载的RNAi对乳腺癌耐药蛋白(BCRP)介导的多药耐药的逆转。方法:构建干扰BCRP mRNA的腺病毒载体,采用细胞毒性实验和外排实验检测pAd-BCRP感染后,耐药细胞后的逆转情况。结果:pAd-BCRP可将B37/MX细胞对米托蒽醌的耐药指数从978降低至75(92.3%逆转)。外排实验结果显示,感染pAd-BCRP和未感染的B37/MX细胞外排1 h后细胞内的米托蒽醌的荧光强度分别为56.4和12.8。结论:pAd-BCRP能显著降低B37/MX细胞中BCRP的表达并抑制BCRP的功能。

OBJECTIVE：To observe the reversing effects of RNAi deliveried by adenovirus on BCRP mediated multidrug resistance.METHODS： RNAi adenovirus targeting breast cancer resistant protein（BCRP） was constructed.The reversal of BCRP was detected by cytotoxicity assays and efflux assays after infection with pAd-BCRP.RESULTS： The multidrug resistance index decreased from 987 to 75（92.3% reverse）.The results of efflux assay showed that,after an hour＇s discharge,the fluorescence intensities were 56.4 and 12.8 in the B37/MX cells infected with adenovirus and without the infection,respectivly.CONCLUSION： RNAi deliveried by adenovrius can decrease the expression of BCRP and reverse MDR mediated by BCRP.