摘要
目的探讨小干扰RNA(small interference RNA,siRNA)影响卵巢癌耐药细胞株MDRl基因的表达并逆转其多药耐药的功能。方法2004年10月至2005年6月于山东省立医院中心实验室采用浓度梯度诱导法建立人卵巢癌阿霉素耐药细胞株OVCAR/AR。根据MDR1基因的编码区域设计了2个含19个碱基的MDR1基因特异性siRNA,脂质体介导下转染OVCAR/AR细胞。RT-PCR分析MDRl mRNA的表达;流式细胞术(FCM)检测P-糖蛋白(P-gp)的表达;MTT法检测OVCAR/AR细胞的多药耐药性。结果MDR1特异性siRNA转染后,OVCAR/AR细胞MDR1mRNA和P-gp的表达减少,分别以转染后48h和72h下降最著;转染mdr1a和mdr1b siR-NA可不同程度地逆转OVCAR/AR细胞对阿霉素、泰素的耐药性,但对非P-gp介导耐药的顺铂无影响。结论阿霉素所致的卵巢癌细胞多药耐药与MDR1基因过度表达有关,体外实验中应用MDR1特异性siRNA能部分逆转OVCAR/AR细胞的多药耐药。
Objective To explore the feasibility of small interference RNA （siRNA） to inhibit MDR1 gene expression and to reverse its muhidrug resistance in ovarian carcinoma drug - resistant cell line. Methods An adriamycin - resistant human ovarian carcinoma cell subline（OVCAR/AR） was established by stepwise inducement from Oct. 2004 to Jun. 2005 in the Central Laboratory of Shandong Provincial Haspital. Two 19bp siRNA （mdrla and mdrlb） which specifically targeted MDR1 gene were designed on base of the coding regions. Transfection of siRNAs into OVCAR/AR cells was performed using liposome transfection reagents. MDR1 mRNA expression level was determined using reverse transcription polymerase chain reaction （ RT - PCR）. Flow cytometry （FCM） was performed to assess the expression of p - glycoprotein （P -gp）. Muhidrug resistance to anticancer agents was evaluated by MTT assay. Results The expression level of MDR1 mRNA and P - gp in OVCAR/AR cells were both reduced by the transfection of MDR1 gene specific siRNAs, and the maximum inhibition was observed 48 and 72 hours after transfection respectively. Transfecting mdrla and mdrlb into OVCAR/AR ceils resulted in different levels of reversal of drug - resistance to Adriamycin and Paclitaxel, but not to non - P - gp mediated MDR drug Cisplatin. Conclusion Muhidrug resistance induced by Adriamycin in ovrian carcinoma cell lines is possibly due to overexpression of MDR1 gene, and it can be partially reversed by MDR1 specific siRNA in vitro.
出处
《中国实用妇科与产科杂志》
CAS
CSCD
北大核心
2006年第1期29-31,I0002,共4页
Chinese Journal of Practical Gynecology and Obstetrics
基金
山东省交通科技计划基金资助(基金编号:2005-Y026)
