CAJ | 학술논문

目的构建靶向于卵巢癌耐药细胞株中高表达的mdr1基因的短发夹状RNA重组质粒,研究其沉默卵巢癌耐药细胞株SKOV3/TAXOL内源mdr1基因对细胞增生的抑制作用.方法运用基因克隆技术构建靶向于mdr1基因的重组质粒pGPU6/GFP/Neo-mdr1并转染至卵巢癌耐药细胞株SKOV3/TAXOL,CCK-8检测对SKOV3/TAXOL细胞增生的抑制作用.结果构建的重组质粒pGPU6/GFP/Neo-mdr1能明显抑制mdr1基因的表达.CCK-8检测结果表明,转染重组质粒pGPU6/GFP/Neo-mdr1的SKOV3/TAXOL靶细胞的抑制率比对照组显著提高(P<0.05).pGPU6/GFP/Neo-mdr1明显抑制SKOV3/TAXOL细胞的增生.细胞凋亡检测显示,pGPU6/GFP/Neo-mdr1转染SKOV3/T后细胞凋亡增加,通过pGPU6/GFP/Neo-mdr1转染SKOV3/T促进凋亡的发生.结论 mdr1基因在耐药的卵巢癌细胞增生与凋亡过程中发挥作用,重组质粒pGPU6/GFP/Neo-mdr1可有效地抑制SKOV3/TAXOL细胞内源mdr1基因的表达和mRNA的转录,并抑制细胞的增生和促进凋亡的发生.
목적 구건파향우란소암내약세포주중고표체적mdr1기인적단발협상RNA중조질립,연구기침묵란소암내약세포주SKOV3/TAXOL내원mdr1기인대세포증생적억제작용.방법 운용기인극륭기술구건파향우mdr1기인적중조질립pGPU6/GFP/Neo-mdr1병전염지란소암내약세포주SKOV3/TAXOL,CCK-8검측대SKOV3/TAXOL세포증생적억제작용.결과 구건적중조질립pGPU6/GFP/Neo-mdr1능명현억제mdr1기인적표체.CCK-8검측결과표명,전염중조질립pGPU6/GFP/Neo-mdr1적SKOV3/TAXOL파세포적억제솔비대조조현저제고(P<0.05).pGPU6/GFP/Neo-mdr1명현억제SKOV3/TAXOL세포적증생.세포조망검측현시,pGPU6/GFP/Neo-mdr1전염SKOV3/T후세포조망증가,통과pGPU6/GFP/Neo-mdr1전염SKOV3/T촉진조망적발생.결론 mdr1기인재내약적란소암세포증생여조망과정중발휘작용,중조질립pGPU6/GFP/Neo-mdr1가유효지억제SKOV3/TAXOL세포내원mdr1기인적표체화mRNA적전록,병억제세포적증생화촉진조망적발생.
Objective To construct the short hairpin RNA recombinant plasmids targeting mdr1 gene which expresses highly in ovarian cancer resistance strain SKOV3/TAXOL to silence endogenefic mdr1 gene expression and investigate the role of mdr1 gene in the development of resistant ovarian cancer. Methods The pGPU6/GFP/Neo-mdr1 were constructed by gene clone technology. The influence on proliferation and apoptosis were investigated by CCK-8 in SKOV3/TAXOL after transfected pGPU6/GFP/Neo-mdr1. Results The expression against mdr1 proteins were inhibited by pGPU6/GFP/Neo-mdr1. The cell proliferation were inhibited after transfected pGPU6/GFP/Neo-mdr1 by CCK-8. The apoptosis were observed in DAB experiments and the apoptosis rate increased. Conclusion mdr1 plays an important role in proliferation of resistant ovarian cancer and the short hairpin RNA of mdr1 can efficiently suppress mdr1 expression and enhance the apoptosis in SKOV3/ATAXOL.