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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 investigate the effect of small interfering RNA (siRNA) on the expression of MDR1 gene in multidrug resistant ovarian cancer cell lines and to reverse its multidrug resistance. Methods From October 2004 to June 2005, the ovarian cancer adriamycin resistant cell line OVCAR / AR was established by concentration gradient method in the central laboratory of Shandong Provincial Hospital. According to the coding region of MDR1, two 19-base-specific MDR1 gene-specific siRNAs were designed and transfected into OVCAR / AR cells by liposome. The expression of MDR1 mRNA was detected by RT-PCR, the expression of P-glycoprotein (P-gp) was detected by flow cytometry (FCM), and the multidrug resistance of OVCAR / AR cells was detected by MTT assay. Results MDR1mRNA and P-gp expression were decreased in OVCAR / AR cells transfected with MDR1-specific siRNA, which were most significantly decreased 48h and 72h after transfection. Transfection of mdr1a and mdr1b siR-NA reversed OVCAR / AR to some extent Cell resistance to doxorubicin, taxol, but non-P-gp-resistant drug-resistant cisplatin no effect. Conclusion Adriamycin-induced ovarian cancer multidrug resistance and MDR1 gene overexpression, in vitro experiments using MDR1-specific siRNA can partially reverse multidrug resistance of OVCAR / AR cells.