資源描述：
《znf278 sirna抑制胃癌細(xì)胞株ags增殖的實(shí)驗(yàn)研究》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。
1、·724ChinJGastroenterol，2013，Vo1．18，No12ZNF278siRNA抑制胃癌細(xì)胞株AGS增殖的實(shí)驗(yàn)研究田筱青孫丹鳳房靜遠(yuǎn)上海交通大學(xué)醫(yī)學(xué)院附屬仁濟(jì)醫(yī)院消化科上海市消化疾病研究所(200001)背景：ZNF278屬C2H2型鋅指蛋白，為參與生長(zhǎng)發(fā)育的重要轉(zhuǎn)錄因子，其異常表達(dá)可能參與腫瘤發(fā)生。目的：研究ZNF278siRNA對(duì)胃癌細(xì)胞株AGS增殖和周期的影響。方法：以蛋白質(zhì)印跡法檢測(cè)正常胃上皮細(xì)胞株GES一1和胃癌細(xì)胞株AGS中ZNF278表達(dá)。構(gòu)建ZNF278siRNA片段，并轉(zhuǎn)染胃癌AGS細(xì)胞，轉(zhuǎn)染陰性對(duì)照siRNA和RPMI1
2、640分別作為陰性對(duì)照組和空白對(duì)照組。以定量RT—PCR和蛋白質(zhì)印跡法分別檢測(cè)ZNF278mRNA和蛋白表達(dá)，利用MTT和細(xì)胞計(jì)數(shù)法檢測(cè)細(xì)胞增殖情況，流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期變化。結(jié)果：胃癌AGS細(xì)胞中ZNF278表達(dá)明顯高于正常胃上皮細(xì)胞GES一1。ZNF278siRNA轉(zhuǎn)染胃癌AGS細(xì)胞后，ZNF278mRNA和蛋白表達(dá)均明顯下調(diào)，而陰性對(duì)照組與空白對(duì)照組無(wú)明顯差異。MITr法示ZNF278siRNA組AGS細(xì)胞增殖率顯著低于陰性對(duì)照組(0．64±0．03對(duì)0．81±0．03，P<0．01)，細(xì)胞計(jì)數(shù)法示細(xì)胞數(shù)量亦顯著降低[(4．I1±0．35)x10’對(duì)(
3、5．78±0．50)x10，P<0．01]，流式細(xì)胞術(shù)顯示ZNF278siRNA組G0／G1期細(xì)胞明顯增加而S期細(xì)胞明顯減少(P<0．05)。結(jié)論：轉(zhuǎn)染ZNF278siRNA可抑制胃癌AGS細(xì)胞增殖，細(xì)胞周期阻滯于G0／G1期。關(guān)鍵詞ZNF278；RNA，小分子干擾；細(xì)胞增殖；細(xì)胞周期；胃腫瘤ExperimentalStudyonSuppressionofCellProliferationinGastricCancerCellLineAGSbyZNF278siRNATIANXiaoqing，SUNDanfeng，F(xiàn)ANGJingyuan．DivisionofG
4、astroenterologyandHepatology，RenJiHospital，SchoolofMedicine，ShanghaiJiaoTongUniversity，ShanghaiInstituteofDigestiveDisease，Shanghai(200001)Correspondenceto：FANGJingyuan，Email：jingyuanfan007@126．cornBackground：ZNF278isanovelKrtippelCys2一His2一typezincfingerprotein．Itisanimportanttrans
5、criptionfactorinvolvedingrowthanddevelopment．AbnormalexpressionofZNF278mightbeinvolvedintumorigenesis．Aims：TostudytheinfluenceofZNF278siRNAoncellproliferationandcellcycleingastriccancercelllineAGS．Methods：ZNF278expressioninhumangastricepithelialcelllineGES一1andgastriccancercelllineA
6、GSwasdetectedbyWesternblotting．ZNF278siRNAwasconstructedandtransfectedintoAGScells．TransfeetionwithnegativesiRNAandRPMI1640wereservedasnegativecontrolsandblankcontrols，respectively．ExpressionsofZNF278mRNAandproteininAGScellsweredeterminedbyquantitativeRT—PCRandWesternblotting，respec
7、tively．CellproliferationwasassessedbyMTTassayandcellcountingassay．Cellcyclewasanalyzedbyflowcytometry．Results：TheexpressionofZNF278washigherinAGScellsthaninGES一1cells．ExpressionsofZNF278mRNAandproteinweresignificantlydecreasedinZNF278siRNAgroupthaninnegativecontrols．Nosignificantdif
8、ferencewasfoundbetw
