Zhang Kaiming, Zhang Ruili, Li Xinhua, Yin Guohua, Niu Xuping
Institute of Dermatology, Taiyuan City Centre Hospital, Shanxi Medical University, No.1 Dong San Dao Taiyuan City, Shanxi Province 030009, P.R. China.
Eur J Dermatol. 2009 Mar-Apr;19(2):141-6. doi: 10.1684/ejd.2008.0618. Epub 2009 Jan 20.
Psoriasis is an inflammatory disease related to dysfunctional immunity. The dysfunctional immunity may influence the haematopoietic microenvironment or haematopoiesis in psoriasis. However, direct evidence is lacking. Our objective was to investigate the proliferation of hematopoietic cells from psoriatic patients and any link between the promoter methylation status of p15 and p21 genes and the colony formation ability of high proliferative potential colony-forming cells (HPP-CFCs). Marrow mononuclear cells were isolated from the bone marrow of psoriatic patients and normal controls by density gradient centrifugalization. Colony forming assays of HPP-CFCs were performed in vitro in methylcellulose semi-solid culture medium. mRNA expression and the promoter methylation status of p15 and p21 genes in HPP-CFCs were studied by semi-quantitative RT-PCR and methylation-specific PCR respectively. In methycellulose semi-solid culture system, the colony count of HPP-CFCs in bone marrow of psoriatic patients was significantly less than that of normal controls. Moreover, significantly lower positive frequencies of promoter methylation and higher transcription levels for p15 and p21 genes were observed in psoriasis in comparison to normal volunteers. The lower promoter methylation of p15 and p21 genes may be an important mechanism for the dysfunctional growth regulation pathways in HPP-CFCs in psoriasis.
银屑病是一种与免疫功能失调相关的炎症性疾病。免疫功能失调可能会影响银屑病患者的造血微环境或造血功能。然而,目前尚缺乏直接证据。我们的目的是研究银屑病患者造血细胞的增殖情况,以及p15和p21基因启动子甲基化状态与高增殖潜能集落形成细胞(HPP-CFCs)集落形成能力之间的关系。通过密度梯度离心法从银屑病患者和正常对照者的骨髓中分离出骨髓单个核细胞。在甲基纤维素半固体培养基中体外进行HPP-CFCs的集落形成试验。分别采用半定量逆转录聚合酶链反应(RT-PCR)和甲基化特异性PCR研究HPP-CFCs中p15和p21基因的mRNA表达及启动子甲基化状态。在甲基纤维素半固体培养体系中,银屑病患者骨髓中HPP-CFCs的集落数明显少于正常对照者。此外,与正常志愿者相比,银屑病患者中p15和p21基因启动子甲基化的阳性频率显著降低,转录水平升高。p15和p21基因启动子甲基化水平降低可能是银屑病患者HPP-CFCs生长调节通路功能失调的重要机制。