银屑病患者骨髓CFUHPP集落形成及集落细胞 p16基因启动子甲基化的研究.DOCVIP

临床医学论文-银屑病患者骨髓CFUHPP集落形成及集落细胞 p16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（CFUHPP）的集落形成能力及集落细胞p16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/GMCSF/IL3/IL6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CFUHPP集落，然后收集集落。提取纯化集落细胞的DNA，经亚硫酸盐修饰后，采用甲基化特异PCR（MSP）检测CFUHPP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CFUHPP集落数显著低于正常对照（t=5.91， p＜0.01），且集落形态较小；正常对照骨髓CFUHPP 集落细胞的p16基因启动子甲基化阳性率较高（66.7%），而银屑病患者骨髓CFUHPP 集落细胞p16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CFUHPP细胞集落形成能力降低；银屑病患者骨髓CFUHPP集落细胞的p16基因甲基化降低可能与其相对较低的CFUHPP集落形成能力密切相关。 【关键词】? 高增殖潜能集落形成单位 ??? CFUHPP Colony Formation of? Bone Marrow Hematopoietic Proginitor Cells in? Psoriatic Patients?? and? Methylation of p16 Gene Promotor in CFUHPP? Colony? Cells ??? Abstract??? This study was purposed to investigate the colony formation of highproliferative potential colonyforming units(CFUHPP) from bone marrowderived hematopoietic cells of psoriatic patients and p16 gene promotor methylation in CFUHPP cells， and to explore the relationship between the colony formation and the methylation status of p16 gene promoter. Bone marrowderived mononuclear cells from psoriatic patients? and normal controls were separated by density gradient centrifugation, and were cultured in methycellulose semisolid culture medium with SCF，GMCSF，IL3 and IL6 for 14 days to measure the colonies of? CFUHPP.? The CFUHPP colony cells were collected and methylation status of p16 gene promoter of?? CFUHPP cell DNA modified? with sodium bisulfite? was detected by the methylationspecific polymerase chain reaction (MSP). The results showed that? in methycellulose semisolid culture system, the number and the size of?? CFUHPP colonies of? bone marrow? of psoriatic patients were all significantly less than that of normal controls, the? positive frequency of p16 gene promoter methylation? in CFUHPP? cells was lower than that in CFUHPP? colony cells of normal controls.? It is concluded that the colony f