银屑病患者骨髓高增殖潜能集落形成细胞P16基因mRNA表达的研究

Study on colony formation capacity and P16 gene mRNA expression of HPP-CFC derived from bone marrow hematopoietic cells of psoriatic patients

目的:研究银屑病患者骨髓高增殖潜能集落形成细胞(HPP-CFC)的集落形成能力及P16基因mRNA表达,探讨银屑病患者骨髓造血干细胞体外增殖活性及原因。方法:密度梯度离心法分离银屑病患者及正常对照骨髓单个核细胞,培养于含人干细胞因子(SCF)、人粒-巨噬细胞系集落剌激因子(GM-CSF)、白介素(IL)-3、IL-6细胞因子组合的甲基纤维素半固体培养基,培养14d时计数HPP-CFC集落,然后收集集落。提取纯化集落细胞的总RNA,应用反转录(RT)-PCR方法检测HPP-CFC集落细胞的P16基因mRNA的表达。结果:①在甲基纤维素半固体培养基中,银屑病患者骨髓HPP-CFC集落数显著低于正常对照组,且集落形态较小;②银屑病患者骨髓HPP-CFC集落细胞的P16基因mRNA表达高于正常对照组,差异有统计学意义。结论:银屑病患者骨髓HPP-CFC集落形成能力降低;银屑病患者骨髓HPP-CFC的P16基因mRNA表达阳性率增高,推测P16基因可能参与了银屑病患者骨髓造血干细胞体外增殖活性异常的发生。

Objective： To investigate the colony formation capacity and P16 gene mRNA expression of high proliferative potential colony forming ceils （HPP-CFC） derived from bone marrow hematopoietic cells of psoriatic patients,and probe into the relationship between the colony formation and P16 promoter mRNA expression. Methods： Bone marrow-derived mononuclear cells from the psoriatics and normal controls were purified by density gradient centrifugation. The bone marrow mononuclear cells were cultured in methylcellulose semi-solid culture medium with SCF,GM-CSF,IL-3 and IL-6 for 14 days. The colony cells were collected and the mRNA expression of P16 gene in colony cells was detected by using reverse transcription-poly- merase chain reaction （RT-PCR）. Results： In methylceilulose semi-solid culture system, the number and size of the colonies of HPP-CFC of psoriatic bone marrow were significantly less than those of the normal control with higher positive frequency of P16 gene mRNA expression compared with the normal control. Conclusion： The higher positive frequency of P16 gene mRNA expression in HPP-CFC may play a role in reducing HPP-CFC colony-forming capability of psoriatic hematopoietic ceils.