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目的探讨小片段RNA干扰抑制乳腺癌耐药蛋白(BCRP)基因及其蛋白表达在裸鼠体内逆转人肝癌组织多药耐药(MDR)的可行性。方法建立裸鼠多药耐药肝细胞癌模型,随机分为A组和B组,A组(对照)注射生理盐水40μl+Lipo-fectamine200010μl,B组注射BCRP基因RNAi质粒pSUPER-BCRP40μl+Lipofectamine200010μl,两组均行瘤内注射1次。5d后,各组均经腹腔注射阿霉素5mg/kg化疗,每5d给药1次,共4次。彩色B超测量肿瘤体积;化疗结束后1周处死裸鼠,RT-PCR及Westernblot法检测各组裸鼠肿瘤组织中BCRP基因mRNA的转录水平及其蛋白的表达水平。结果B组每次化疗后肿瘤体积均明显缩小;除第1次化疗外,其余各次化疗后肿瘤体积均小于A组;化疗结束后,B组与A组相比,肿瘤组织中BCRP基因mRNA的转录水平和BCRP蛋白的表达水平均明显降低。结论BCRP基因RNAi质粒pSUPER-BCRP可有效降低裸鼠肝癌组织BCRP基因mRNA的转录水平及其蛋白的表达水平,在一定程度上逆转MDR,为从基因水平逆转MDR提供了初步的实验依据。
Objective To investigate the feasibility of small interfering RNA (RNAi) repression of breast cancer resistance protein (BCRP) gene and its protein expression in reversing human multidrug resistance (MDR) in human hepatocellular carcinoma in nude mice. Methods The model of multi-drug resistant hepatocellular carcinoma in nude mice was established and randomly divided into A group and B group. A group (control) was injected with 40μl of saline and Lipo-fectamine200010μl. B group was injected with BCRP RNAi plasmid pSUPER-BCRP40μl + Lipofectamine200010μl, Group were intratumoral injection 1 times. After 5 days, all groups were given intraperitoneal injection of doxorubicin 5mg / kg chemotherapy, once every 5d, a total of 4 times. The tumor volume was measured by color B ultrasound. One week after the chemotherapy, the nude mice were sacrificed. The transcriptional level of BCRP mRNA and the expression of BCRP mRNA in the nude mice were detected by RT-PCR and Western blot. Results The volume of tumor in group B was significantly reduced after each chemotherapy. The volume of tumor in group B was less than that in group A except for the first chemotherapy. After chemotherapy, the expression of BCRP mRNA The transcription level and BCRP protein expression levels were significantly lower. Conclusion The RNAi plasmid pSUPER-BCRP of BCRP gene can effectively reduce the transcript level of BCRP mRNA and protein in HCC of nude mice, and reverse the MDR to a certain extent. It provides a preliminary experimental evidence for reversing MDR at gene level.