反义寡聚脱氧核苷酸抑制卵巢癌细胞株SKOV3/mdr1中MDR1基因的表达  

Inhibition of MDR1 gene expression in ovarian carcinoma cell line SKOV3/mdr1 by antisense oligodeoxynucleotide

作　　者：金滢[1] 潘凌亚[1] 童英[1] 张毅[2] 张双喜[2] 吴英[2] 毛宁[2] 杨秀玉[1] 

机构地区：[1]中国医学科学院中国协和医科大学北京协和医院妇产科,北京100730 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《基础医学与临床》2001年第2期127-130,共4页Basic and Clinical Medicine

摘　　要：多药耐药基因 (MDR1)编码的P -糖蛋白 (Pgp ,P - 170 )是导致卵巢癌化疗失败的重要原因之一。本研究将针对MDR1基因的硫代反义寡聚脱氧核苷酸导入卵巢癌耐药细胞株SKOV3/mdr1,观察其对该细胞株P170表达的影响。反义寡聚脱氧核苷酸 (MDR1-AS)通过阳离子脂质体介导或单纯转染的方式导入SKOV3/mdr1细胞 ,将正义寡聚脱氧核苷酸 (MDR1-S)同样导入细胞为对照。经以脂质体介导的 1.6 μmol/LMDR1-AS转染后 ,Pgp阳性的细胞百分数从 10 0 %降至 48.7% ,经过 16和 10 μmol/L两个浓度的单纯MDR1-AS转染后 ,Pgp阳性细胞的百分数也从 10 0 %分别降至 5 2 .6 %和 86 .7%。同时 ,利用RT -PCR检测了MDR1mRNA水平的变化 ,以β -actin作为内对照 ,发现MDR1mRNA水平在MDR1-AS转染后降至内对照的 6 0 %左右 ,而用MDR1-S转染的细胞与SKOV3/mdr1细胞比无明显变化。所以导入MDR1基因硫代反义寡聚脱氧核苷酸可以部分抑制SKOV3/mdr1细胞中MDR1基因的表达。One of the most important sources, which result in failure in ovary cancer treatment, is P-glycoprotein, which is coded by multidrug resistance gene (mdr1). To overcome the problem of multidrug resistance, the effectiveness of phosphrothioate antisense oligodeoxynucleotides(MDR1-AS)in suppressing MDR1 gene expression in drug-resistance SKOV3/mdr1 cell line was investigated in this study. MDR1-AS was transfected into SKOV3/mdr1 cell mediated by lipofectamine or not with MDR1-S treatment group as control. The percentage of Pgp + cells was decreased from 100% to 48.7% by 1.6μmol/L MDR1-AS plus lipofectamine treatment and from 100% to 52.6% or 86.7% by 16μmol/L or 10μmol/L MDR1-AS treatment respectively. MDR1 mRNA level detected by RT-PCR was also diminished to about 60% of β-actin after MDR1-AS plus lipofectamine treatment with β-actin as internal control. So MDR1 expression in SKOV3/mdr1 cell line was partially inhibited after MDR1-AS treatment in the mRNA and protein level.

关 键 词：反义寡核苷酸 卵巢癌 多药耐药 基因治疗 基因表达 

分 类 号：R737.31[医药卫生—肿瘤] R730.5[医药卫生—临床医学]