大鼠肝卵圆细胞向肝癌细胞分化过程中microRNA的变化-论文.pdf

《大鼠肝卵圆细胞向肝癌细胞分化过程中microRNA的变化-论文.pdf》由会员上传分享，免费在线阅读，更多相关内容在应用文档-天天文库。

1、第24卷第7期China中Jo国ur现na代lo医fM学od杂er志nMedicineVo1．24No．72014年3月Mar．2014文章编号：1005—8982(2014)07—0001—06·论著·大鼠肝卵圆细胞向肝癌细胞分化过程中microRNA的变化术许荣华，何冬雷，孟津，夏立平，王健z(1．海南医学院附属医院胃肠肿瘤外科，海南海口570102；2．南方医科大学附属花都医院普外科，广东广州510800)摘要：目的利用生物芯片技术分析大鼠肝卵圆细胞分化为肝癌细胞过程中微小RNA(microRNA，miRNA)的表达变化，筛选调控分化的micr

2、oRNAs，探讨肝癌发生的起源机制。方法用化学致癌剂3’一Me—DAB诱发SD大鼠肝卵圆细胞增生，采用密度梯度离心法分离、纯化肝卵圆细胞，行体外培养使其恶性分化并进行鉴定。提取分化过程中的microRNA进行microRNA基因芯片杂交，获得卵原细胞恶性分化过程中microRNA表达谱。同时利用种植模型及大鼠Y染色体特异性PCR技术，验证卵圆细胞可恶性分化为肝癌细胞。结果①激光扫描共聚焦显微镜显示，肝卵圆细胞胞浆和胞膜表达干细胞标志Thy一1及c—kit；②通过基因微阵列分析，卵原细胞恶性分化过程中有22个miRNA的改变显著，其中19个表达上调，3

3、个表达下调；③卵原细胞种植模型的大鼠肝癌组织具有SRY基因的电泳条带。结论特定microRNA可能对大鼠肝卵圆细胞分化为肝癌细胞过程起到关键作用。关键词：肝卵圆细胞；分化；microRNA；微小RNA芯片中图分类号：R～332文献标识码：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．andmicroRNAwasisolatedfromthetotalRNA．MicroarrayanalysisofmicroRNAexpressionwasper—formedtodetectthedifferentexpressionlevelsofmicroRNAamongth

7、eindicatedtimepoints．RatsYchromosomespecificpolymerasechainreactiontechniquewasusedtofindouttheevidencethatHOCscoulddifferentiatedintolivercancercells．【Results】LaserscanningconfocalmicroscopyindicatedpositiveexpressionofstemcellsmarkersThv一1andC—kitincytoplasmandmembraneofHOCs．

8、Atotalof1210miRNAswereidentifiedandamongthem22werediff