临床医学论文 银屑病患者骨髓CFUHPP集落形成及集落细胞 p16基因启动子甲基化的研究.doc

《临床医学论文 银屑病患者骨髓CFUHPP集落形成及集落细胞 p16基因启动子甲基化的研究.doc》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。

1、银屑病患者骨髓CFUHPP集落形成及集落细胞pl6基因启动子甲基化的研究作者：张瑞丽，牛旭平,李新华，张开明，尹国华【摘要】本研究检测银屑病患者骨髓高增殖潜能集落形成单位(CFUHPP)的集落形成能力及集落细胞pl6基因启动子甲基化状态，并探讨两者之间的关系。收集了24例银屑病患者及正常对照者骨髓，采用密度梯度离心法分离单个核细胞，并于含SCF/GMCSF/IL3/IL6细胞因子组合的甲基纤维素半固体培养液中，培养14天计数CFUHIT集落，然后收集集落。提取纯化集落细胞的DA,经亚硫酸盐修饰后

2、，采用甲基化特异PCR(MSP)检测CFUHPP集落细胞的P16基因启动子甲基化状态。结果发现：在甲基纤维素半固体培养基中，银屑病患者骨髓CFUIIPP集落数显著低于正常对照(t=5.91,pVO.Ol),且集落形态较小；正常对照骨髓CFUHPP集落细胞的pl6基因启动子甲基化阳性率较高(66.7%),而银屑病患者骨髓CFUHPP集落细胞pl6基因启动子甲基化阳性率(37.5%)低于正常人。结论：银屑病患者骨髓CFUHPP细胞集落形成能力降低；银屑病患者骨髓CFUHIT集落细胞的pl6基因中基化降

3、低可能与其相对较低的CFUHPP集落形成能力密切相关。【关键词】高增殖潜能集落形成单位CFUHPPColonyFormationofBoneMarrowHematopoieticProginitorCel1sinPsoriaticPeitientsandMethylationofpl6GenePromotorinCFUIIPPColonyCel1sAbstractThisstudywaspurposedtoinvestigatethecolonyformationofhighproliferati

4、vepotentialcolonyformingunits(CFUHIT)frombonemarrowderivedhematopoieticcellsofpsoriaticpatientsandpl6genepromotormethylationinCFUHPPcells,andtoexploretherelationshipbetweenthecolonyformationandthemethylationstatusofpl6genepromoter.Bonemarrowderivedcon

5、trolswereculturedinCSF,IL3HPP.Themononuclearcel1sfrompsoriaticpatientsandnormalseparatedbydensitygradientcentrifugation,andweremethycellulosesemisolidculturemediumwithSCF,GMandIL6for14daystomeasurethecoloniesofCFUCFUHPPcolonycellswerecollectedandmethy

6、lationstatusofpl6genepromoterofCFUHPPcellDNAmodifiedwithsodiumbisulfitewasdetectedbythemethylationspecificpolymerasechainreaction(MSP).TheresultsshowedthatinmethycelluloseCFUHPPsemisolidculturesystem,thenumberandthesizeofpositiveHPPcellswascontrols.It

7、CFUIIPPfromcoloniesofbonemarrowofpsoriaticpatientswereallsignificantlylessthanthatofnormalcontrols,thefrequencyofpl6genepromotermethylationinCFUlowerthanthatinCFUHPPcolonycellsofnormalisconcludedthatthecolonyformationcapabi1ityofbonemarrowhematopoieti

8、cprogenitorcel1sinpsoriaticpatientsis1owerthanthatinnormalcontrols,andthelowerpositivefrequencyofP16genepromotermethylationinCFUHPPcellsperhapscloselycorrelatedwithlowerCFUHPPcolonyformingcapabi1ity.Keywordspsoriasis;highproliferativepotential