Role of WAVE1 in K562 leukemia cells invasion

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1、万方数据虫堡堂速堂苤查2Q塑生垒旦筮!Q鲞箜垒翅垡堕!』坐!些堕丛：生堕12Q塑：坠!：!Q。盟!：璺WAVEl对K562细胞侵袭能力的影响及其机制研究贺钰磊曹励之杨明华赵明一俞燕许望琼【摘要】目的研究WASP家族富含脯氨酸同源蛋白1(WAVEl)对l(562细胞侵袭的影响及其机制。方法免疫荧光观察WAVEI与基质金属蛋白-2(MMP-2)在细胞中的分布。利用pcDNA3．1一WAVEl重组真核表达质粒转染K562细胞，将WAVEl基因特异性小片段干扰RNA(WAVEIsiRNA)转染K562细胞，Transwell法检测细胞侵袭能力；实时PCR和W

2、esternblot检测转染前后WAVEl及MMP-2的表达。结果①WAVEI与MMP-2主要表达于l(562细胞的细胞膜上且两者有共定位。②与对照组K562细胞相比，转染pcDNA3．1-WAVEI24h及48h后K562细胞MMP-2mRNA表达分别增加295％和198％，蛋白表达水平分别增加80％和23％；转染特异性WAVElsiRNA24h及48h后K562细胞MMP-2mRNA表达分别下降8l％和28％，蛋白表达分别下调36％和53％。③与对照组K562细胞相比转染pcDNA3．1-WAVEI后细胞侵袭能力增强，而干扰WAVEI表达后l(5

3、62细胞侵袭能力减弱。结论WAVEl与MMP-2在K562细胞可能具有协同作用；WAVEl参与了K562细胞的侵袭转移过程，其机制町能与调控MMP-2的表达相关。【关键词】WASP家族富含脯氨酸同源蛋白l；基质金属蛋白酶；K562细胞系；RNA干扰；白血病浸润RoleofWAVElin酗62Ieukemiacellsinvasionanditsmechanism而陋玩．玩．甜0H-zhi．翻^临Ming—hua，zHAoMing-yi，YUYan，XUWang—qiong．DepartmentofPediatrics．XiangyaHospitalo

4、fCentralSouthUniversity。Changsha4iO008．ChinaCorrespondingauthor：CAf)Li．zhi．Email：caolizhi318@hotmail．com【Abstract】ObjectiveToinvestigateroleofWASPfamilyverprolinhomologousprotein1(wAVEl)inK562leukemiacellinvasionandi协mechanism．MethodsImmunofluoresencemethedwasusedtodetectthedis

5、tributionofWAVElandMMP-2iilthecells．K562cellsweretransfectedwithPeDNA3．1一WAVElm-constructedplasmidorwithspecificsiRNAtoWAVEIgene．TheinvasionabilitvofK562cellswasexaminedbvTranswellassay．TheexpressionlevelofWAVElandMMP-2iUK562cellsWasassayedbyreal—timePCRandWestemblot．Results(DW

6、AVElandMMP-2mainlyexpressedandCO．10calizedinthecellmembrane；(国24hand48haftertransfectedwithpcDNA3．1一WAVEl，theMMP-2mRNAlevelinK562cellsincreasedby295％and198％whileitsproteinincreasedby80％and23％respectivelyascomparedwithcontroll(562cells．Atthesalnetimepointoftertransfectedwithspec

7、ificsiRNA．theMMP-2mRNAleveldecreasedby8l％and28％．anditsproteindecreasedby36％and53％respectivelyascomparedwithcontr01．(固’11lein—vasionabilityofl巧62cellsWasenhancedaftertransfectedwithpcDNA3．1一WAVElanddepressedaftertrans-fectedwiththespecificsiRNA．ConclusionTheCO．10calizationofWAVE

8、landMMP-2inK562ceHssug-geststheycoordinateinfunctions：