载体表达小干扰RNA逆转卵巢癌的多药耐药

The Effect of Vector Based Small Interfering RNAs in Reversal of Multidrug Resistance in Ovarian Cancer Cell Line

目的:探讨载体表达的小干扰RNA(smallinterferingRNA,siRNA)逆转卵巢癌细胞多药耐药的可行性。方法:浓度梯度诱导法建立人卵巢癌阿霉素耐药细胞株OVCAR/AR;脂质体介导将MDR1特异性siRNA的表达载体(pSN/mdr1a和pSN/mdr1b)转染OVCAR/AR细胞;实时定量RT-PCR检测MDRlmRNA的表达;流式细胞术检测P-gp的表达,罗丹明试验检测P-gp的药物转运功能;MTT法检测OVCAR/AR细胞对化疗药的抵抗性。结果:转染pSN/mdr1a和pSN/mdr1b后,OVCAR/AR细胞的MDR1mRNA和P-gp的表达均显著下降,P-gp的转运功能减少,OVCAR/AR细胞对阿霉素、泰素的耐药性逆转。结论:载体表达的siRNA可持久有效地抑制卵巢癌耐药细胞MDR1mRNA和P-gp的表达,并逆转其多药耐药。

Objective： To explore the feasibility of using vector-based small Interfering RNA （siRNA） to reverse the multidrug resistance in drug-resistant ovarian cancer cell line. Methods： An adriamycin-resistant human ovarian cancer cell subline （OVCAR/AR） was established by stepwise inducement. Transfection of MDR1 mRNA specific siRNA expressing plasmids （pSN/mdrla and pSN/ mdrlb） into OVCAR/AR cells was performed using liposome transfection reagents. MDR1 mRNA expression level was quantified using real-time reverse transcription polymerase chain reaction （real time RT-PCR）. Flow cytometry （FCM） was performed to assess the expression of p-glycoprotein （P-gp）. The / transporting function of P-gp was determined by Rhodamine assay. Multidrug resistant to anticancer a- gents was evaluated by MTF assay. Results： Basal MDR1 mRNA and P-gp expression level in drugresistant cell line OVCAR/AR was higher than that in its parent cell line OVCAR-3. After transfection with pSN/mdrla and pSN/mdrlb, the expression level of MDR1 mRNA and P-gp protein was reduced. The transporting function of P-gp to drugs was reduced simultaneously. Treatment of OVCAR/AR cells with pSN/mdrla and pSN/mdrlb resulted in a different level of reversal of drug-resistantance to P-gp mediated drugs Adriamycin and Paclitaxel. Conclusion： Vector-based MDR1 specific siRNA can in- hibit the expression levels of both MDR1 mRNA and P-gp continuously and effectively, thereby to sensitize their Adriamycin-resistant ovarian cancer cells.