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1、第24卷第7期China中Jo國ur現(xiàn)na代lo醫(yī)fM學(xué)od雜er志nMedicineVo1．24No．72014年3月Mar．2014文章編號(hào)：1005—8982(2014)07—0001—06·論著·大鼠肝卵圓細(xì)胞向肝癌細(xì)胞分化過程中microRNA的變化術(shù)許榮華，何冬雷，孟津，夏立平，王健z(1．海南醫(yī)學(xué)院附屬醫(yī)院胃腸腫瘤外科，海南?？?70102；2．南方醫(yī)科大學(xué)附屬花都醫(yī)院普外科，廣東廣州510800)摘要：目的利用生物芯片技術(shù)分析大鼠肝卵圓細(xì)胞分化為肝癌細(xì)胞過程中微小RNA(microRNA，miRNA)的表達(dá)變化，篩選調(diào)控分化的micr
2、oRNAs，探討肝癌發(fā)生的起源機(jī)制。方法用化學(xué)致癌劑3’一Me—DAB誘發(fā)SD大鼠肝卵圓細(xì)胞增生，采用密度梯度離心法分離、純化肝卵圓細(xì)胞，行體外培養(yǎng)使其惡性分化并進(jìn)行鑒定。提取分化過程中的microRNA進(jìn)行microRNA基因芯片雜交，獲得卵原細(xì)胞惡性分化過程中microRNA表達(dá)譜。同時(shí)利用種植模型及大鼠Y染色體特異性PCR技術(shù)，驗(yàn)證卵圓細(xì)胞可惡性分化為肝癌細(xì)胞。結(jié)果①激光掃描共聚焦顯微鏡顯示，肝卵圓細(xì)胞胞漿和胞膜表達(dá)干細(xì)胞標(biāo)志Thy一1及c—kit；②通過基因微陣列分析，卵原細(xì)胞惡性分化過程中有22個(gè)miRNA的改變顯著，其中19個(gè)表達(dá)上調(diào)，3
3、個(gè)表達(dá)下調(diào)；③卵原細(xì)胞種植模型的大鼠肝癌組織具有SRY基因的電泳條帶。結(jié)論特定microRNA可能對(duì)大鼠肝卵圓細(xì)胞分化為肝癌細(xì)胞過程起到關(guān)鍵作用。關(guān)鍵詞：肝卵圓細(xì)胞；分化；microRNA；微小RNA芯片中圖分類號(hào)：R～332文獻(xiàn)標(biāo)識(shí)碼：AMicroRNAdifferentialexpressionprofileduringdifferentiationofratovalcellstowardhepatocellularcarcinomaXURong—hua，HEDong-lei，MENGJing，XIALi-ping，WANGJian(1．Depa
4、rtmentofOncologySurgery,theAffiliatedHospitalofH~nanMedicalCollege，H~kou，Hainan570102，P．R．China；2．DepartmentofGeneralSurgery,HuaduHospital,SouthernMedicalUniversity,Guangzhou，Guangdong510800,P．R．China)Abstract：【0bjective】ToexploretheexpressionprofileofmicroRNAsduringdifferentia
5、tionofrathepaticovalcells(HOCs1towardhepatocellularcarcinomaSOastoprovideabasisforfurtherexploringtheroleofmiRNAsinthemolecularmechanismthatlcadstotheoccu1Tenceanddevelopmentoflirecancer．IMethods】ProliferationofratH0Cswasinducedbychemicalcarcinogen．3一Me—DAB．HOCswereisolatedByPe
6、rcolldensitygradienteentrifu—gationmethod．~llowedbycontinuouscultivationinvitro．TotalRNAwasextractedfromHOCsduringcellsdiffer—entiation．a(chǎn)ndmicroRNAwasisolatedfromthetotalRNA．MicroarrayanalysisofmicroRNAexpressionwasper—formedtodetectthedifferentexpressionlevelsofmicroRNAamongth
7、eindicatedtimepoints．RatsYchromosomespecificpolymerasechainreactiontechniquewasusedtofindouttheevidencethatHOCscoulddifferentiatedintolivercancercells．【Results】LaserscanningconfocalmicroscopyindicatedpositiveexpressionofstemcellsmarkersThv一1andC—kitincytoplasmandmembraneofHOCs．
8、Atotalof1210miRNAswereidentifiedandamongthem22werediff
