HIF-2α基因與人肺腺癌A549細(xì)胞株對順鉑耐藥性的關(guān)系-論文.pdf

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1、772ChineseJournalofClinicalMedicine，2013．Vo1．20，No．6中國臨床醫(yī)學(xué)2013年12月第20卷第6期論著HIF一2基因與人肺腺癌A549細(xì)胞株對順鉑耐藥性的關(guān)系吳倩’袁凱錢成王勇”童繼春袁衛(wèi)東¨(1．南京醫(yī)科大學(xué)附屬常州第二人民醫(yī)院血液腫瘤科，胸心外科，江蘇常州213003；2．復(fù)旦大學(xué)附屬中山醫(yī)院胸外科，上海200032)摘要目的：探討小干擾RNA(smallinterferingRNA，siRNA)沉默缺氧誘導(dǎo)因子2(hypoxiainduciblefactor2alpha，HIF一2a)基因表達(dá)后，人肺腺癌A549細(xì)胞株對順鉑耐藥性的變化

2、。方法：構(gòu)建靶向HIF一2d基因的siRNA真核表達(dá)載體并轉(zhuǎn)染A549細(xì)胞，采用Real—timePCR檢測HIF-2dmRNA的表達(dá)，采用Westernblotting檢測HIF-2c~及多藥耐藥基因1(multidrugresistanee一1，Mdr一1)蛋白的表達(dá)，采用MTT法檢測順鉑處理后細(xì)胞的存活率。結(jié)果：在HIF2asiRNA轉(zhuǎn)染A549細(xì)胞后24、48h后，HIF-2d在mRNA和蛋白水平的表達(dá)與空白對照組相比均顯著下調(diào)(均P<0．01)，Mdr一1蛋白的表達(dá)水平也明顯降低(均P<()．05)。與空白對照組相比，HIF2dsiRNA組細(xì)胞對順鉑的敏感性明顯增加，各時間點(diǎn)的I

3、C值均顯著降低，差異有統(tǒng)計學(xué)意義(均P<()．01)。結(jié)論：靶向HIF一2“的siRNA表達(dá)載體能夠有效抑制HIF一2a基因的表達(dá)，從而逆轉(zhuǎn)A549細(xì)胞對順鉑的化療耐藥性。關(guān)鍵詞肺腺癌；化療耐藥；缺氧誘導(dǎo)因子2a；小干擾RNA；多藥耐藥基因1中圖分類號R734．2文獻(xiàn)標(biāo)識碼ARelationshipbetweenHIF一2“GeneandChemotherapyResistanceofLungAdenocarcinomaA549CellstoCis—platinWUQianL4NKaiQIANChengW_ANGYongT0NGJichun礎(chǔ)．NWP—dong1．DepartmentofH

4、ematologyandOncology，DepartmentofThoracicSurgery，ChangzhouSecondPeople’sHospital，NanjingMedicalUniversity，Changzhou213003，China；2．DepartmentofThoracicSurgery，ZhongshanHospital，F(xiàn)udanUniversity，Shanghai200032，ChinaAbstractObjective!Toinvestigatetheeffectofsilencinghypoxiainduciblefactor-2alpha(HIF一2

5、a)geneexpressionbysmallinterferingRNA(siRNA)onchemotherapyresistanceofA549cellstocisplatin．Methods：ThesiRNAeukaryoticexpressionvectorstargetingHIF一2agenewereconstructedandtransfectedintoA549cells．TheexpressionofHIF2amRNAwasdetectedbyreal—timePCR．TheexpressionofHIF一2aandmultidrugresistance1(Mdr-1)p

6、roteinsweredetectedbyWesternblotting．MTTassaywasusedtoexplorethesurvivalrateofA549cellsunderthetreatmentofcisplatin．Results：AftertransfectionofHIF一2asiRNAintoA549cellsfor24and48h，theexpressionlevelsofHIF2dmRNAandproteinwerebothdown—regulatedsignificantlycomparedwiththoseintheblankgroup(allP

7、Meanwhile，theproteinexpressionlevelofMdr一1alsodecreasedsignificantly(allP<0．05)．ThesensitivityofA549cellstocisplatinwassignificantlyincreasedinHIF2“siRNAtransfectiongroupcomparedwiththatintheblankgroup．TheIC5{)va