c-FLIP介导HIF-1α/GLUT-1信号通路在银屑病角质形成细胞增殖中的机制研究

Overexpressed c-FLIP can disturb degradation of HIF-1α in lung cancer cell line,and HIF-1α is very important for cell proliferation and angiogenesis. Our previous data showed that both HIF-1α and c-FLIP protein highly expressed in psoriatic skin compare to normal skin; and the HIF-1α expression level correlated with expression of GLUT-1 the downstream signal of HIF-1α.However, the mechanism of HIF-1α upregulation in psoriatic skin still unclear.In this study, we try to overexpress or inhibit c-FLIP expression in keratinocyte from biopsies by Lenti-virus expression system, and then to observe HIF-1α protein expression and translocation;to check HIF-1α downstream signal, and to analysis relationship between HIF-1α signal and keratinocyte proliferation. Additionally,we try to explore the interaction mechanism between HIF-1α and c-FLIP by gene reporter analysis, co-immunoprecipition and point mutation technicals. This study first bring the idea of c-FLIP upreguate keratinocyte proliferation though HIF-1α/GLUT-1 pathway, which involve in pathogenesis of psoriasis.

基于HIF-1α在细胞增殖中的重要作用，及部分肿瘤组织中高表达的c-FLIP可以干扰HIF-1α的代谢等相关文献知识，我们前期发现HIF-1α、c-FLIP在银屑病皮损中表达均较正常皮肤组织高，且c-FLIP的表达与HIF-1α相关；HIF-1α的表达与下游靶基因GLUT-1及细胞增殖呈正相关。但银屑病皮损中HIF-1α表达上调的原因及机制并不清楚。因此本项目拟通过慢病毒基因转染及RNA干扰技术，诱导或阻断角质形成细胞c-FLIP的表达，观察其对HIF-1α及其下游靶基因的影响及其由此所致的银屑病角质形成细胞增殖能力的改变。拟用报告基因分析、免疫共沉淀及点突变等技术探索c-FLIP影响HIF-1α的作用机制及其位点。本课题首次提出c-FLIP通过影响HIF-1α/GLUT-1通路从而影响角质形成细胞增殖等生物学行为，并对其机制进行探讨。