上调mir-143表达对caski人宫颈鳞癌细胞系增殖与凋亡的影响

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1、临床与实验病理学杂志JClinExpPathol2010Apr；26(2)·223·上调miR一143表达对CasKi人宫颈鳞癌细胞系增殖与凋亡的影响王晓玫，高利昆，成志强，彭全洲，胡锦涛，许静，徐洲稳摘要：目的探讨miR-143对CasKi宫颈鳞癌细胞增殖与凋亡活性影响。方法应用荧光定量RT—PCR法检测miR一143在人宫颈鳞癌组织中表达水平，以及miR一143在CasKi细胞株中表达水平与变化；人工合成miR一143模拟体转染CasKi宫颈鳞癌细胞系，采用流式细胞技术、MTT等实验方法探讨上调miR一143表达对CasKi癌细

2、胞增殖活性与凋亡影响。结果宫颈鳞癌组织与宫颈鳞癌细胞株CasKi中miR一143表达水平较正常宫颈组织明显下调，其中2例鳞癌组织未能检测出miR一143表达。miR一143mimics转染组miR一143表达显著高于空白对照组；MTF检测结果表明miR一143mimics转染组中细胞增殖抑制明显，且抑制效应自转染48h开始显现。细胞周期检测结果显示miR143mimics转染组Go／G期细胞明显增多，s期细胞减少，组问差异有统计学意义(P<0．05)。荧光TUNEL检测结果亦显示miR一143mimics转染组细胞凋亡率增加(9．4

3、54-2．31)％，高于阴性对照组(4．03±1．35)％与空白对照组(3．85±1．61)％，组间差异具有统计学意义(P<0．05)。结论上调miR一143表达可抑制宫颈鳞癌细胞增殖活性，影响宫颈鳞癌细胞周期和诱导宫颈鳞癌细胞凋亡。关键词：宫颈肿瘤；鳞状细胞癌；miR一143；CasKi细胞；细胞增殖；凋亡中图分类号：R737．33；R363．2文献标识码：A文章编号：1001—7399(2010)02—0223—05Effectsofup--regulatedexpressionofmicroRNA-143oncellproli

4、ferationandapoptosisofCasKicellsWANGXiao—mei，GAOLi—kun，CHENGZhi—qiang，PENGQuan-zhou，HUJin—tao，XUJing，XUZhou—wen(DepartmentofPathology，theSecondAfiliatedHospital，nUniversity／ShenzhenPeople’sHospital，Shenzhen518020，China)Abstract：PurposeToinvestigatetheeffectsofup—regula

5、tedexpressionofmicroRNA一143(miR一143)oncellproliferationandapop—tosisofcervicalsquamouscarcinomacelllinesCasKi．MethodsSyntheticmiR一143mimicswastransfectedintoCasKicelllinebyli—pofectamine．TheexpressionofmiR一143incervicalsquamouscellcarcinomatissuesandtheexpressionchange

6、ofmiR一143intheCas—KicelllinewasmeasuredbyrealtimeRT—PCR．Cellulargrowthandproliferationcapacitywereassayedbythemethylthiazolyltetrazo—lium(M．rr)assayandfloweytometry(FCM)．TheapoptoticratewastestedbyTUNELassay．ResultsTheexpressionofmiR一143inCasKicellandcervicalsquamousce

7、llcarcinomatissueswasdown—regulated，incontrasttonormalcervicaltissue，andtwocarcino—masampleswerenonexpressionofmiR一143．Comparedwiththatofblankcontrolgroup，miR一143expressioninthemiR一143mimicsgroupcellincreasedsignificantly；thecellgrowthinhibitionofmiR一143mimicsgroupwasd

8、emonstratedfor48hoursaftercellstreatedwithtransfectionbyM

9、ITI1results；FCManalysisshowedthatcellsatGo／GIphaseinthemiR一