小分子干扰RNA逆转胃癌SGC7901/VCR细胞mdr1介导的多药耐药  

THE REVERSAL EFFECT ON mdr1 GENE-MEDIATED MULTIDRUG RESISTANCE IN GASTRIC CANCER SGC7901/VCR CELLS BY SMALL INTERFERING RNA

作　　者：高福莲[1] 刘书漫[1] 吴景兰[1] 张钦宪[1] 

机构地区：[1]郑州大学基础医学院组织学胚胎学教研室河南省分子医学重点学科开放实验室,郑州450052

出　　处：《解剖学报》2006年第1期57-61,共5页Acta Anatomica Sinica

基　　金：河南省重大科技攻关项目(0222031300)

摘　　要：目的研究小分子干扰RNA(siRNA)逆转胃癌多药耐药细胞亚系SGC7901/VCR mdr1介导的多药耐药效应。方法根据mdr1cDNA已知序列,设计并体外转录2条含21个核苷酸的siRNA(mdr1si2631和mdr1si3071),转染SGC7901/VCR细胞,用RT-PCR检测mdr1基因mRNA的表达,免疫组织化学检测P-gp的表达,流式细胞仪检测阿霉素在细胞内的蓄积,MTT法检测细胞对阿霉素的敏感性。结果siRNA转染SGC7901/VCR细胞48 h后,mdr1基因mRNA和P-gp的表达水平下降,细胞内阿霉素积累量增加,对阿霉素敏感性的相对逆转率各达79.59%及59.98%。结论siRNA可逆转SGC7901/VCR细胞mdr1介导的多药耐药。Objective To investigate the reversal effect on mdr1 gene mediated muhidrug resistance in gastric cancer SGC7901NCR cells by small interfering RNA （siRNA）. Methods Two siRNAs （mdr1si2631 and mdr1si3071 ） specifically targeting mdr1 gene were designed and synthesized by transcription in vitro. The siRNA duplexes were used to transfect into the gastric cancer SGC7901/VCR cells. The expression levels of mdr1 mRNA and P-gp were detected by RT-PCR and immunohistochemistry respectively. The accumulation of intracellular adriamycin（ ADR）was examined by flow cytometry and the cell sensitivity to ADR was demonstrated by M3W. Results The expression level of mdr1 mRNA treated by siRNAs for 48 hours was decreased in the SGC7901/VCR cells. The mdr1 RT-PCR product in the transfected mdrlsi2631 SGC7901/VCR cells could hardly been found, similar to its parental SGC7901 cells, the ratio of mdr1 and β-actin in the control SGC7901/VCR group was 1.05 ±0.10, the transfected mdrl si3071 group was 0.16 ±0.03 （ P 〈 0.001 ） and the transfected group with transfection reagent alone was 0.92± 0.01 （ P 〉 0.05 ）. The RT- PCR results showed that the mdrl mRNA expression level in the mdrl si2631 group decline more obviously than that in the mdrlsi3071 group, near by the level in its parental SGC7901 cells. The pogp immunoreactivity （IR）in brownish-colored granules was located on the cell membrane.The P-gp IR became weaker in the SGC7901/VCR cells treated by siRNAs for 48 hours and the P-gp expression level in both transfected siRNA groups was decreased.The values of adriamycinspecific fluorescence intensity and the positive rates of intracellular ADR in both transfected siRNA groups were increased.The relative reversal efficiency of the SGC7901/VCR cells to ADR detected by MTT was 79.59 % in mdr1 si2631 group and 59.98 % in mdrlsi3071 group respectively. Conclusion siRNA could reverse mdrl gene mediated multidrug resistance in gastric cancer SGC7901/VCR cells.

关 键 词：小分子干扰RNA MDR1基因 SGC7901/VCR细胞 多药耐药 逆转录聚合酶链反应 噻唑蓝法 人 

分 类 号：R735.2[医药卫生—肿瘤]