目的:通过筛选早年白发患者和正常黑发者毛发中差异表达的基因,为研究毛发黑素合成及转运的生理病理机制提供线索。方法:留取门诊确诊的早年白发患者毛发,应用退火控制引物(ACP)初步筛选差异表达的候选基因,实时定量PCR法进一步确认候选基因的表达水平;设计合成筛选出的Slc7a11和小眼相关转录因子(Mitf)的siRNA后,转染入正常人黑素细胞系PIG1,在H2O2诱导下,通过Annexin-V FITC检测其凋亡。结果:ACP筛选及RT-PCR验证结果提示,Slc7a11和Mitf基因表达水平在早年白发患者和正常人间有较大差异(P〈0.01);降低这两种基因的表达后,PIG1的凋亡率较正常组升高。结论:Slc7a11和Mitf基因不仅在正常毛发色素合成中发挥着作用,同时也参与着氧化应激诱导下黑素细胞凋亡的过程,可能在一些色素相关性疾病的发生发展中起着关键的作用。
Objective: To investigate the various genes between premature greying of hair patients and healthy subjects by screening,which implied a way for understanding the pathogenesis of melanin genesis and transport.Methods: Hair of premature greying patients were collected,and annealing control primers(ACP) were preliminarily used to screen for candidate genes,then the expression level of these genes were confirmed through Real-time PCR.siRNA were used to transfect into PIG1 cell lines,then the apoptosis rates were analyzed by AnnexinV-FITC,under H2O2-induced oxidative stress.Results: There was significance in both Slc7a11 and Mitf genes expression,between patients and healthy ones(P0.01).When these two genes were down regulated,the apoptosis rate of PIG1 increased upon oxidative stress.Conclusion: Slc7a11 and Mitf are major genetic regulators of melanin pigment in hair and provide more evidence for the further investigation of pigmentation disorder,including the process of oxidative stress-induced melanocytes apoptosis.