论文部分内容阅读
目的研究沉默KIF4A基因对人卵巢癌SKOV3细胞迁移能力的影响。方法检索针对KIF4A基因的小干扰RNA(siRNA),构建shRNA表达质粒,转染人卵巢癌细胞SKOV3,通过G418筛选建立稳定低表达KIF4A的SKOV3细胞株。采用蛋白质免疫印记(Western blot)法检测shRNA对KIF4A蛋白分子的敲除效率。并通过细胞迁移实验评价低表达KIF4A对SKOV3细胞迁移能力的影响。结果成功筛选出稳定低表达KIF4A的细胞株,选择干扰效率较高的细胞株进行实验。Transwell细胞迁移实验显示,低表达KIF4A的细胞株的迁移率较阴性对照组增长近3倍(P<0.05)。结论 KIF4A低表达,可增强人卵巢癌细胞SKOV3的迁移能力。
Objective To study the effect of silenced KIF4A gene on the migration of human ovarian cancer SKOV3 cells. Methods Small interfering RNA (siRNA) against KIF4A gene was screened and shRNA expression plasmid was constructed. The human ovarian cancer cell line SKOV3 was transfected into SKOV3 cells. The SKOV3 cell line stably expressing KIF4A was screened by G418. The knockdown efficiency of KIF4A protein by shRNA was detected by Western blot. The effect of low expression of KIF4A on migration of SKOV3 cells was evaluated by cell migration assay. Results The cell lines stably expressing KIF4A were selected successfully, and the cell lines with high interference efficiency were selected for experiments. Transwell cell migration assay showed that the cell migration rate of KIF4A-overexpressing cells increased nearly three-fold (P <0.05) compared with the negative control group. Conclusion The low expression of KIF4A enhances the migration of SKOV3 in human ovarian cancer cells.