Nek2基因沉默对卵巢癌SKOV3细胞侵袭能力的影响  

Silencing Nek2 via RNAi suppresses invasiveness in ovarian cancer SKOV3 cells

作　　者：范婷婷[1] 唐良萏[1] 

机构地区：[1]重庆医科大学附属第一医院妇产科,重庆400016

出　　处：《第三军医大学学报》2012年第15期1514-1517,共4页Journal of Third Military Medical University

基　　金：教育部博士培养基金(200806310001);重庆医科大学附属第一医院医学科学基金(YXJJ2009-05)~~

摘　　要：目的研究RNA干扰Nek2表达对卵巢癌SKOV3细胞侵袭能力的影响以及相关的分子机制。方法设计合成3条针对Nek2基因的siRNA,转染至SKOV3细胞中,采用Real-time RT-PCR和Western blot技术筛选出抑制效率最高的Nek2-siRNA序列,用Transwell方法检测转染该序列的SKOV3细胞侵袭能力的改变,并用Western blot方法检测Nek2-siRNA转染后SKOV3细胞中MMP-2、MMP-9和TIMP-1蛋白表达的变化。结果①RNA干扰序列2对SKOV3细胞中Nek2的抑制效果最明显;②与未转染组和阴性对照组相比,转染了Nek2-siRNA的SKOV3细胞侵袭能力明显下降(P<0.01);③Western blot检测表明,与未转染组和阴性对照组相比,Nek2-siRNA转染48 h后SKOV3细胞中MMP-2、MMP-9的表达明显下调,而TIMP-1的表达明显上调(P<0.01)。结论 Nek2-siRNA能有效得沉默SKOV3细胞中Nek2的表达,通过下调MMP-2、MMP-9的表达,上调TIMP-1的表达,从而抑制SKOV3细胞的侵袭转移。Objective To determine the effect of silencing NIMA-related kinase 2（Nek2） via RNAi on the invasive capacity of ovarian cancer SKOV3 cells.Methods Three pairs of siRNA were designed according to the sequence of Nek2 gene and then synthesized chemically.In 48 h after transfection of the siRNAs into the SKOV3 cells,real-time RT-PCR and Western blotting were performed to detect the expression levels of Nek2 mRNA and protein so as to screen the most effective siRNA.The capacity of invasion were evaluated by Transwell chamber test.Western blotting was used to determine the protein levels of MMP-2,MMP-9 and TIMP-1 after Nek2-siRNA transfection.Results Real-time RT-PCR and Western blotting revealed that Nek2-siRNA notably down-regulated Nek2 expression at both mRNA and protein levels.The cell number of invasion in SKOV3 cells was significantly decreased compared to the control groups（P〈0.01）.Western blotting revealed that Nek2-siRNA transfection up-regulated the expression level of TIMP-1,down-regulated the expression levels of MMP-2 and MMP-9 in SKOV3 cells significantly（P〈0.01）.Conclusion Nek2-siRNA transfection can effectively inhibits Nek2 expression,restrains the invasion capacities of ovarian cancer SKOV3 cells through up-regulating TIMP-1 and down-regulating MMP-2 and MMP-9.

关 键 词：NIMA相关激酶 RNA干扰 卵巢癌 侵袭 

分 类 号：R394.3[医药卫生—医学遗传学] R737.31[医药卫生—基础医学]