中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (6): 407-410.

• 论著 • 上一篇    下一篇

银屑病患者骨髓造血干细胞差异表达基因的筛选与鉴定

田畔1,于浈2,张振颖3,彭洪菊4,刘晓明5   

  1. 1. 大连医科大学附属第一医院(现工作单位：龙岩市中心医院）
    2. 大连医科大学附属第一医院（现工作医院：大连市皮肤病医院）
    3. 大连医科大学附属第一医院皮肤科教研室
    4. 大连医科大学附属第一医院
    5. 大连医科大学附属第一医院皮肤科
  • 收稿日期:2010-09-19 修回日期:2010-10-12 出版日期:2011-06-15 发布日期:2011-06-02
  • 通讯作者: 刘晓明 E-mail:Lxm_1956@yahoo.com.cn
  • 基金资助:

    国家自然基金

Screening for and identification of differentially expressed genes in hemopoietic stem cells from patients with psoriasis

  • Received:2010-09-19 Revised:2010-10-12 Online:2011-06-15 Published:2011-06-02
  • Contact: liu xiaoming E-mail:Lxm_1956@yahoo.com.cn

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摘要：

目的 探讨骨髓造血干细胞在银屑病发病中的作用。方法 分选纯化人骨髓造血干细胞，构建寻常性银屑病患者与正常人对照者骨髓造血干细胞差异表达的基因文库，对经菌液PCR及斑点杂交筛选出的阳性克隆进行基因测序、基因鉴定及功能分析，并检测银屑病患者及健康对照者骨髓造血干细胞中干扰素γ及TMSB10胸腺素的mRNA水平。结果 正向库中鉴定出干扰素γ，酪氨酸磷酸酶，SUMO1激活酶等9个基因。反向库中鉴定出TMSB10胸腺素等8个基因。银屑病患者骨髓造血干细胞的干扰素γ mRNA水平为正常对照组的47.5倍，而正常人对照者TMSB10胸腺素mRNA水平是患者表达量的22.6倍。结论 银屑病患者造血干细胞中干扰素γ、胸腺素等17个基因表达的异常可能与银屑病的发病有关。

关键词: 差异表达基因

Abstract:

Objective To investigate the roles of hemopoietic stem cells （HSCs） in the pathogenesis of psoriasis. Methods HSCs were isolated from the bone marrow of a patient with psoriasis vulgaris and a normal human control. Forward- and reverse-subtracted cDNA libraries were constructed by using suppression subtractive hybridization （SSH） technique between HSCs from the patient and control. Bacterial PCR and dot hybridization were performed to screen for positive clones followed by gene sequencing, identification and functional analysis. Real time quantitative PCR was carried out to measure the mRNA expression of interferon-γ and thymosin 10 （TMSB10）. Results Nine genes were screened from the forward-subtracted cDNA library which encoded interferon-γ, tyrosine phosphatase, SUMO1 activase, etc, and 8 genes including the TMSB10-encoding gene were screened from the reverse-subtracted cDNA library. The relative expression level of interferon-γ in HSCs from the patient was 47.5 times that in HSCs from the control, while the level of TMSB10 from the control was 22.6 times that from the patient. Conclusion The abnormal expression of 17 genes which encode interferon-γ, thyrosin, and so on, may be involved in the development of psoriasis.

Key words: Differentially expressed genes