RNAi沉默卵巢癌耐药细胞株SKOV3／TAXOL内源mdr1基因对细胞增生的影响

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，

The proliferative effect on silencing of endogenetic mdr1 gene expression by RNA interference in ovarian cancer resistant strain SKOV3/TAXOL

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.