【摘要】 目的探讨Wip1在不同卵巢癌细胞系中的表达情况,筛选出Wip1强表达的细胞系,为进一步研究Wip1在卵巢癌细胞系中的作用机制奠定基础。方法应用RT-PCR及Western blot检测正常卵巢上皮细胞及卵巢癌细胞株SKOV3、CAOV3、A2780、ES2中Wip的表达情况,最终筛选出SKOV3作为实验细胞系。在卵巢癌SKOV3细胞中沉默Wip基因观察其生物学行为的变化。结果 Wip1mRNA表达在SKOV3细胞中明显高于正常上皮细胞(P<0.01),在CAOV3、A2780、ES2细胞中也均高于卵巢正常上皮细胞(P<0.05)。Wip1蛋白表达在SKOV3、A2780、ES2细胞中明显高于正常上皮细胞(P<0.01),在CAOV3细胞中也高于卵巢正常上皮细胞(P<0.05)。转染siRNA-2的SKOV3细胞中Wip1蛋白表达最少,Wip1-siRNA-2转染SKOV3细胞后,Wip1的表达随着其浓度的增加呈减少趋势,在浓度为80 nmol/L达到最低值;而作用时间为48 h Wip1蛋白表达也达到最低(P<0.01)。结论 Wip1在SKOV3、CAOV3、A2780、ES2细胞中均呈过表达,在SKOV3细胞中表达水平最高,Wip1-siRNA转染SKOV3细胞在特定浓度和作用时间时沉默效果最好,为进一步阐述其分子机制提供了理论基础。更多还原

【Abstract】 Objective To investigate the expression levels of Wip1 in different ovarian cancer cell lines to screen out the cell line with the highest expression of Wip1,so as to lay the foundation for further research on the action mechanism of Wip1 in ovarian cancer cell lines.Methods The expression levels of Wip1 mRNA and protein in normal ovarian epithelial cells and ovarian cancer cell lines-SKOV3,CAOV3,A2780 and ES2 were detected by RT-PCR and Western-blot.SKOV3 was screened out as an experimental cell line.Wip1 gene in ovarian cancer SKOV3 cells was silenced to observe the changes of its biological behaviors.Results The expression levels of Wip1 mRNA in SKOV3 cells were significantly higher than those in normal epithelial cells(P<0.01),which in CAOV3,A2780 and ES2 cells were significantly higher than those in normal epithelial cells(P<0.05).The expression levels of Wip1 protein in SKOV3,A2780,ES2 and CAOV3 cells were significantly higher than those in normal epithelial cells(P<0.01 or P<0.05).The expression levels of Wip1 protein in SKOV3 cells that were transfected with siRNA-2 were the least.After Wip1-siRNA-2 was transfected into SKOV3 cells,the expression levels of Wip1 were decreased with the increase of Wip1 concentration,which were the least at a concentration of 80 nmol/L,and the expression levels of Wip1 protein were the least at 48 hours after treatment.It was demonstrated that Wip1-siRNA transfected SKOV3 cells had the most obvious effects at a specific concentration and reaction time(P<0.01).Conclusion The overexpression of Wip1 is found in SKOV3,CAOV3,A2780,ES2 cells,its expression levels are the highest in SKOV3 cells,and Wip1-siRNA transfected SKOV3 cells has the best silencing effects at a specific concentration and reaction time,which provides theoretical basis for further elaboration of its molecular mechanism.更多还原