siRNA干扰降低ARK5或Gab2表达对LN-229细胞侵袭的影响及作用机制  

SiRNA Interference Reducing ARK5 or Gab2 Is Associated with the Influence of Invasion and Its Mechanism in LN-229 Cells

作　　者：孙磊[1] 李小龙[1] 田红艳[1] 刘雨清[1] 

机构地区：[1]潍坊医学院病理学教研室,山东潍坊261053

出　　处：《潍坊医学院学报》2014年第1期9-11,共3页Acta Academiae Medicinae Weifang

摘　　要：目的探讨siRNA干扰降低ARK5或Gab2表达对LN-229细胞侵袭的影响及作用机制。方法采用小RNA干扰技术转染LN-229细胞株,并用Western blot分别检测转染前和瞬时转染后ARK5和Gab2蛋白表达。通过体外侵袭实验检测转染后侵袭能力的变化。ARK5或Gab2下降后明胶酶谱分别检测MMP-2和MMP-9蛋白表达。结果 转染ARK5,Gab2及SCR质粒的LN-229细胞分别称SiARK5/LN-229,SiGab2/LN-229和SCR/LN-229。转染后,与SCR/LN-229及LN-229相比,SiARK5/LN-229中ARK5蛋白表达降低,SiGab2/LN-229中Gab2蛋白表达降低。ARK5或Gab2降低的LN-229细胞侵袭并穿透Matrigel膜基质的细胞数量均比SCR/LN-229少(P<0.01)。与SCR/LN-229细胞相比,SiARK5/LN-229和SiGab2/LN-229中MMP-2,MMP-9蛋白表达均降低。结论 应用小RNA干扰技术降低ARK5或Gab2的表达使LN-229细胞侵袭力降低,同时MMP-2和MMP-9表达降低,提示ARK5和Gab2表达降低可通过调节MMP-2和MMP-9的表达影响LN-229细胞的侵袭。Objective To investigate the influence of invasion anti its mechanism in 1.N-229 cells by siRNA interference reduc- ing ARK5 or Gab2. Methods Western blot was used to detect the expression of ARK5 and Gab2 in LN-229 cells. SiRNA plasmid was used to transfeet I,N-229 cells anti then Western blot was used to analyze protein expression of ARK5 and Gab2. Matrigel invasion assay was used to detect the variation of invasiveness in I,N-229 cells being transfected. Zymography were detected MMP-2, MMP-9 protein expression after ARK5 or （,ab2 expression had decreased. Results Transfeeted ARK5,Gab2 and SCR plasmid LN-229 eells were called SiARK5/LN-229, Si（,ab2/LN-229 anti SCR/LN-229 cells. The protein expression of ARK5 or Gab2 in I.N-229 cells that had been transfected by ARK5 or （Cab2 was decreased. （Compared with SCR/I.N-229 cell, the quantity of LN-229 eells after downregulation of ARK5 or Gab2 whieh invaded and penetrated Matrigel were both decreased（ P 〈 0.01 ） as well as the protein expression of MMP-2 and MMP-9 were decreased significantly. Conclusion Silencing of ARK5 or Gab2 impairs glioma cell invasion and the expression of ARK5 or Gab2 in glioma is significantly related with the expression of MMP-2 anti MMP-9,suggesting that ARK5 or Gab2 can play a key role in the invasiveness of LN-229 by adjnsting MMP2 and MMP-9 expression.

关 键 词：胶质瘤 ARK5 Gab2 LN-229细胞 小RNA干扰 侵袭 

分 类 号：R730.23[医药卫生—肿瘤]