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1、·論羞·JMedRes,Dec2013,Vo1.42No.12RNA沉默Pinl表達(dá)對(duì)乳腺癌細(xì)胞增殖、侵襲和凋亡的影響溫媛媛楊志強(qiáng)摘要目的應(yīng)用RNA干擾技術(shù)沉默Pinl表達(dá)后,觀察其對(duì)乳腺癌細(xì)胞增殖、侵襲和凋亡的影響。方法采用脂質(zhì)體介導(dǎo)法將Pinl干擾片段PinlsiRNA和對(duì)照片段controlsiRNA轉(zhuǎn)染入乳腺癌細(xì)胞系MDA—MB一23l后,利用RT—PCR和Westernblot法檢測(cè)Pinl基因在mRNA和蛋白水平的表達(dá)情況,并應(yīng)用MTT法、Transwell法和AnnexinV—FITC/PI試劑盒及流式細(xì)胞儀技術(shù)分析檢測(cè)Pinl對(duì)MDA—MB一23l細(xì)胞增
2、殖、侵襲和凋亡的影響。結(jié)果Pinl在乳腺癌細(xì)胞系中高表達(dá)。與未處理組及轉(zhuǎn)染對(duì)照片段controlsiRNA組相比,沉默Pinl表達(dá)后,PinlmRNA(P<0.05)和蛋白(P<0.05)表達(dá)均明顯下降,細(xì)胞生長(zhǎng)增殖能力[P>0.05(第1天),P<0.O1(第2—4天)]減弱,細(xì)胞侵襲數(shù)目(6.61±0.53,P<0.05)減少,細(xì)胞凋亡率(31.5%4-3.06%,P<0.05)顯著增加。結(jié)論沉默Pin1表達(dá)后可抑制乳腺癌細(xì)胞增殖和侵襲,促進(jìn)乳腺癌細(xì)胞凋亡。關(guān)鍵詞乳腺癌Pinl增殖侵襲凋亡Down—regulationofPinlbyRNAInferenceonCe
3、llProliferation,InvasionandApoptosisinBreastCancerCells.WenYuanyuan,YangZhiqiang.ZhoushanHospital,Zhejiang316000,ChinaAbstractObjectiveToinvestigatetheeffectsofPinlreductiononcellproliferation,invasionandapoptosisofbreastcancerMDA—MB一231cellsbysilencingPinlgeneusingRNAinterferencetechniq
4、ue.MethodsMDA—MB一231cellsweretransfectedwithPinlsiRNAandcontrolsiRNAusingLipofeetamine2000.TheexpressionofSIAH1mRNAandproteinweredetectedbyRT—PCRandWest—ernblot.ThecellproliferationwasanalyzedbyMTTandthecelIinvasionwastestedbyTranswel1.AnnexinV—FITC/PIreagentkitandFlowcytometryassaywereu
5、sedtoanalyzecellapoptosis.ResultsTheexpressionofPinlwashigherinbreastcancerceilsthaninnor—realmammaryepithelialcells.ComparedwithuntreatedandthecellstransfectedwithcontrolsiRNA,thelevelofPinlmRNA(P<0.05)andprotein(P<0.05)weredecreasedinthecellstransfectedwiththePinlsiRNA.Thecellgrowthrat
6、elP>0.05(day1),P<0.O1(day2—4)]wasslower,thenumbersofcellinvasion(6.61±0.53,P<0.05)weredecreased,andtheapoptoticrate(31.5%4-3.06%.P<0.05)wasincreasedinMDA—MB一23lcellstransfectedwithPinlsiRNA.ConclusionKonckdownofPinlwithPinlsiRNAmaysuppresscellproliferationandinvasion,promotecellapoptosis
7、inbreastcancercells.KeywordsBreastcancer;Pinl;Proliferation;Invasion;Apoptosis乳腺癌是一種常見的惡性腫瘤。由于其早期發(fā)達(dá),并參與多種腫瘤的發(fā)生發(fā)展過(guò)程,被稱為腫瘤發(fā)病時(shí)的臨床癥狀隱匿,所以多數(shù)患者發(fā)現(xiàn)時(shí)已處于進(jìn)生發(fā)展的催化分子,與腫瘤的增殖失控、血管形成、惡展期,預(yù)后也較差,因此尋找有助于早期診斷的生物性侵襲有關(guān)。。然而Pinl在乳腺癌發(fā)生發(fā)展中的學(xué)指標(biāo)對(duì)于乳腺癌診斷有著重要的意義。此外,這些具體機(jī)制至今還不是很清楚。本研究分析Pinl在乳指標(biāo)也可以為我們治療乳