摘要目的研究针对MDR1基因的反义寡核柑酸对卵巢癌耐药细胞株SKOV3/mdr1耐药性的逆转作用.方法将MDR1基因导入卵巢癌细胞株SKOV3作为耐药细胞的模型,命名为SKOV3/mdr1.以脂质体lipofectamine介导将MDR1基因的反义寡核柑酸导入SKOV3/mdr1,同样将正义寡核柑酸导入上述细胞为对照.利用反转录聚合酶链反应(RT-PCR)检测细胞内MDR1基因mRNA的量.MDR1基因表达的产物P糖蛋白及其功能分别以流式细胞仪及罗丹明123排出实验检测.细胞的耐药性由MTT实验及细胞集落培养来检测.结果经MDR1基因的反义寡核柑酸转染后,SKOV3/mdr1细胞内MDR1基因mRNA的量降至内对照β-actin mRNA的60%.P糖蛋白阳性细胞的相对百分数由100%降至52.6%(P<0.01).细胞内罗丹明123由9.1增至33.8%,对泰素的耐药性降至SKOV3/mdr1细胞的58%.与SKOV3/mdr1细胞相比,在特定的药物浓度范围内,经MDR1基因的反义寡核柑酸转染后,SKOV3/mdr1细胞较未转染细胞对泰素和阿霉素的抗性细胞集落分别减少了8.6±0.8和3.1±0.6倍.结论 SKOV3/mdr1细胞经MDR1基因的反义寡核柑酸转染后部分抑制了MDR1基因在mRNA水平的转录及在蛋白水平的表达,从而增加了该细胞对化疗的敏感性.

abstractsObjective　To investigate the effect of multidrug resistance gene 1 (mdr1) antisense oligodeoxynucleotides (ODNs) on reversing multidrug resistance in the drug resistant ovarian carcinoma cell line SKOV3/mdr1. Methods　The ovarian carcinoma cell line SKOV3 transducted with a human multidrug resistance gene (mdr1) served as the drug resistant model (SKOV3/mdr1). The mdr1 antisense ODNs was transfected into SKOV3/mdr1 cells while mediated by lipofectamine. Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression and the amount of the mdr1 mRNA in the cells. The positive rate and function of the mdr1 gene product P-glycoprotein (Pgp) in the mdr1 antisense ODNs treated SKOV3/mdr1 cells were determined by flow cytometry and rhodamine 123 efflux. Drug resistance in the SKOV3/mdr1 cell line was observed by MTT assay and cell colony culture. Results　The mdr1 mRNA level was decreased to about 60% of that of β-actin after mdr1 antisense ODNs treatment. The Pgp positive rate of mdr1 antisense ODNs treated SKOV3/mdr1 cells decreased from 100% to 52.6% (P<0.01). The intracellular rhodamine 123 retention was increased from 9.1% to 33.8% (P<0.01). The chemoresistance to taxol decreased to 58% of SKOV3/mdr1 with mdr1 antisense ODN treatment. Compared with SKOV3/mdr1 cells in the control group, under a certain range of drug concentrations, the number of drug resistance colonies in mdr1 antisense ODNs treated SKOV3/mdr1 cells for taxol and doxorubicin decreased by 8.6±0.8 fold and 3.1±0.6 fold, respectively. Some non-specific functions during oligodeoxyncleotide treatment was also detected. Conclusion　mdr1 expression in the SKOV1/mdr1 cell line was partially inhibited after mdr1 antisense ODNs treatment at the mRNA and protein level, increasing the chemotherapy sensitivity of this drug resistant ovarian carcinoma cell line.