The role of MDR1 gene in volume-activated chloride currents in pigmented ciliary epithelial cells

(1 Laboratory of Cell Biology, 2 Department of Physiology, Guangdong Medical College, Zhanjiang  524023, China;
3 Cardiff School of Biosciences, Cardiff University, Cardiff,  UK)

Abstract: 

The role of multidrug resistance  (MDR1)  gene in the activation of volume-activated chloride currents in bovine pigmented ciliary epithelial (PCE) cells was investigated by the patch-clamp technique, the antisense approach, the immunofluorescent technique and the confocal microscopy. PCE cells express P-glycoprotein (P-gp, the product of MDR1 gene).  An MDR1 antisense oligonucleotide suppressed MDR1 expression (93% reduction of P-gp immunofluorescence), delayed the activation of a volume-activated chloride current (latency prolonged by 109%), reduced the activation rate by 62% and decreased the peak value of the current by 56%. The transfection reagent lipofectin and the mismatch control oligonucleotide did not significantly affect the current. The data indicate that the volume-activated chloride current is associated with the endogenous expression of MDR1 gene in PCE cells.

Key words:  

epithelial cells; chloride channels; oligonucleotides, antisense;
 gene expression; genes;  MDR

MDR1基因在睫状体色素上皮细胞容积激活性氯电流中的作用

陈丽新1,3,王立伟2,3， Tim JACOB3

（广东医学院 1细胞生物学研究室，2生理教研室， 湛江  524023；
  3Cardiff School of Biosciences, Cardiff University, Cardiff, UK）

用膜片钳、反义寡核苷酸、免疫荧光及激光共聚焦显微镜等技术,  研究MDR1基因在牛睫状体色素上皮(pigmented ciliary epithelial,  PCE)细胞容积激活性氯电流中的作用。PCE细胞表达MDR1基因产物-P糖蛋白(P-gp)。反义MDR1寡核苷酸抑制MDR1基因的表达(P-gp免疫荧光减少93％)，延缓容积激活性氯电流的出现(潜伏期延长109％)，并导致激活率降低62％及电流峰值减小56%。而核酸转染剂阳离子脂质体和非配对性的寡核苷酸对电流没有显著性影响。上述观察结果表明睫状体色素上皮细胞容积激活性氯电流与内源性MDR1 表达有关。

关键词: 上皮细胞;  氯通道; 寡核苷酸; 反义; 基因表达; 基因; MDR
学科分类号: Q25

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