[1]范婷婷,唐良萏.Nek2基因沉默对卵巢癌SKOV3细胞侵袭能力的影响[J].第三军医大学学报,2012,34(15):1514-1517.
 Fan Tingting,Tang Liangdan.Silencing Nek2 via RNAi suppresses invasiveness in ovarian cancer SKOV3 cells[J].J Third Mil Med Univ,2012,34(15):1514-1517.
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Nek2基因沉默对卵巢癌SKOV3细胞侵袭能力的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第15期
页码:
1514-1517
栏目:
论著
出版日期:
2012-08-15

文章信息/Info

Title:
Silencing Nek2 via RNAi suppresses invasiveness in ovarian cancer SKOV3 cells
作者:
范婷婷唐良萏
重庆医科大学附属第一医院妇产科
Author(s):
Fan Tingting Tang Liangdan
Department of Obstetrics and Gynecology, First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China
关键词:
NIMA相关激酶RNA干扰卵巢癌侵袭
Keywords:
Nek2 small interfering RNA ovarian cancer cell invasiveness
分类号:
R394.3; R73-37; R737.31
文献标志码:
A
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。
Abstract:
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.

参考文献/References:

范婷婷, 唐良萏. Nek2基因沉默对卵巢癌SKOV3细胞侵袭能力的影响[J].第三军医大学学报,2012,34(15):1514-1517.

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更新日期/Last Update: 2012-07-26