Pan L, Tong Y, Jin Y, Zhou S, Zhang Y, Yang X, Mao N
Department of Obstetrics and Gynecology, Chinese Academy of Medical Sciences, Peking Union Medical College Hospital, Beijing 100730, China.
Chin Med J (Engl). 2001 Sep;114(9):929-32.
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.
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.
The mdr1 mRNA level was decreased to about 60% of that of beta-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.
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.
研究多药耐药基因1（mdr1）反义寡脱氧核苷酸（ODNs）对耐药卵巢癌细胞系SKOV3/mdr1多药耐药性的逆转作用。
将转导人多药耐药基因（mdr1）的卵巢癌细胞系SKOV3作为耐药模型（SKOV3/mdr1）。在脂质体介导下，将mdr1反义ODNs转染到SKOV3/mdr1细胞中。采用逆转录-聚合酶链反应（RT-PCR）检测细胞中mdr1 mRNA的表达及含量。通过流式细胞术和罗丹明123外排实验检测mdr1反义ODNs处理的SKOV3/mdr1细胞中mdr1基因产物P-糖蛋白（Pgp）的阳性率和功能。采用MTT法和细胞集落培养观察SKOV3/mdr1细胞系的耐药性。
mdr1反义ODNs处理后，mdr1 mRNA水平降至β-肌动蛋白的约60%。mdr治疗后的SKOV3/mdr1细胞的Pgp阳性率从100%降至52.6%（P<0.01）。细胞内罗丹明123的保留率从9.1%提高到33.8%（P<0.01）。mdr1反义ODN处理后，对紫杉醇的化疗耐药性降至SKOV3/mdr1的58%。与对照组SKOV3/mdr1细胞相比，在一定药物浓度范围内,mdr1反义ODNs处理的SKOV3/mdr1细胞对紫杉醇和阿霉素的耐药集落数分别减少了8.6±0.8倍和3.1±0.6倍。还检测到寡脱氧核苷酸处理过程中的一些非特异性作用。
mdr1反义ODNs处理后，SKOV1/mdr1细胞系中mdr1在mRNA和蛋白水平的表达受到部分抑制，提高了该耐药卵巢癌细胞系的化疗敏感性。
