潍坊医学院学报
濰坊醫學院學報
유방의학원학보
JOURNAL OF WEIFANG MEDICAL COLLEGE
2014年
1期
9-11
,共3页
孙磊%李小龙%田红艳%刘雨清
孫磊%李小龍%田紅豔%劉雨清
손뢰%리소룡%전홍염%류우청
胶质瘤%ARK5%Gab2%LN-229细胞%小RNA干扰%侵袭
膠質瘤%ARK5%Gab2%LN-229細胞%小RNA榦擾%侵襲
효질류%ARK5%Gab2%LN-229세포%소RNA간우%침습
LN-229%siRNA interference%ARK5%Gab2%Invasiveness
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。
目的：探討siRNA榦擾降低ARK5或Gab2錶達對LN-229細胞侵襲的影響及作用機製。方法採用小RNA榦擾技術轉染LN-229細胞株，併用Western blot分彆檢測轉染前和瞬時轉染後ARK5和Gab2蛋白錶達。通過體外侵襲實驗檢測轉染後侵襲能力的變化。 ARK5或Gab2下降後明膠酶譜分彆檢測MMP-2和MMP-9蛋白錶達。結果轉染ARK5，Gab2及SCR質粒的LN-229細胞分彆稱SiARK5/LN-229，SiGab2/LN-229和SCR/LN-229。轉染後，與SCR/LN-229及LN-229相比，SiARK5/LN-229中ARK5蛋白錶達降低，SiGab2/LN-229中Gab2蛋白錶達降低。 ARK5或Gab2降低的LN-229細胞侵襲併穿透 Matrigel 膜基質的細胞數量均比SCR/LN-229少（P＜0．01）。與SCR/LN-229細胞相比，SiARK5/LN-229和SiGab2/LN-229中MMP-2，MMP-9蛋白錶達均降低。結論應用小RNA榦擾技術降低ARK5或Gab2的錶達使LN-229細胞侵襲力降低，同時MMP-2和MMP-9錶達降低，提示ARK5和Gab2錶達降低可通過調節MMP-2和MMP-9的錶達影響LN-229細胞的侵襲。
목적：탐토siRNA간우강저ARK5혹Gab2표체대LN-229세포침습적영향급작용궤제。방법채용소RNA간우기술전염LN-229세포주，병용Western blot분별검측전염전화순시전염후ARK5화Gab2단백표체。통과체외침습실험검측전염후침습능력적변화。 ARK5혹Gab2하강후명효매보분별검측MMP-2화MMP-9단백표체。결과전염ARK5，Gab2급SCR질립적LN-229세포분별칭SiARK5/LN-229，SiGab2/LN-229화SCR/LN-229。전염후，여SCR/LN-229급LN-229상비，SiARK5/LN-229중ARK5단백표체강저，SiGab2/LN-229중Gab2단백표체강저。 ARK5혹Gab2강저적LN-229세포침습병천투 Matrigel 막기질적세포수량균비SCR/LN-229소（P＜0．01）。여SCR/LN-229세포상비，SiARK5/LN-229화SiGab2/LN-229중MMP-2，MMP-9단백표체균강저。결론응용소RNA간우기술강저ARK5혹Gab2적표체사LN-229세포침습력강저，동시MMP-2화MMP-9표체강저，제시ARK5화Gab2표체강저가통과조절MMP-2화MMP-9적표체영향LN-229세포적침습。
Objective To investigate the influence of invasion and its mechanism in LN-229 cells by siRNA interference reduc-ing ARK5 or Gab2.Methods Western blot was used to detect the expression of ARK 5 and Gab2 in LN-229 cells.SiRNA plasmid was used to transfect LN-229 cells and then Western blot was used to analyze protein expression of ARK 5 and Gab2.Matrigel invasion assay was used to detect the variation of invasiveness in LN-229 cells being transfected.Zymography were detected MMP-2,MMP-9 protein expression after ARK5 or Gab2 expression had decreased .Results Transfected ARK5,Gab2 and SCR plasmid LN-229 cells were called SiARK5/LN-229, SiGab2/LN-229 and SCR/LN-229 cells.The protein expression of ARK5 or Gab2 in LN-229 cells that had been transfected by ARK5 or Gab2 was decreased.Compared with SCR/LN-229 cell,the quantity of LN-229 cells after downregulation of ARK5 or Gab2 which 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 ARK 5 or Gab2 in glioma is significantly related with the expression of MMP-2 and MMP-9,suggesting that ARK5 or Gab2 can play a key role in the invasiveness of LN-229 by adjusting MMP2 and MMP-9 expression.
