lncRNA01535通过JAK/STAT3通路调节毛囊干细胞增殖和分化的作用研究

Effects of LncRNA01535 on regulating the proliferation and differentiation of hair follicle stem cells through JAK/STAT3 signaling pathway

目的探讨长链非编码RNA01535(lncRNA01535)通过Janus激酶/信号转导与转录激活子3(JAK/STAT3)通路调节毛囊干细胞(HFSCs)增殖和分化的作用。方法设HFSCs细胞组、lncRNA01535过表达组(lncRNA01535 mimics组)、lncRNA01535低表达组(lncRNA01535 inhibitor组);各组细胞培养72 h后,CCK-8实验测定细胞存活水平,红油O染色测定细胞分化水平,流式细胞仪测定细胞凋亡及细胞周期,实时荧光定量PCR(RT-qPCR)法及蛋白免疫印迹(Western blot)法测定lncRNA01535、JAK、STAT3 mRNA及蛋白表达水平。结果与HFSCs细胞组比较,lncRNA01535 mimics组、细胞存活率、细胞分化率升高,细胞凋亡率、G_(1)期细胞占比降低(P<0.05);lncRNA01535 inhibitor组、细胞存活率、细胞分化率降低,细胞凋亡率、G_(1)期细胞占比升高(P<0.05);与lncRNA01535 mimics组比较,lncRNA01535 inhibitor组细胞存活率、细胞分化率降低,细胞凋亡率、G_(1)期细胞占比升高(P<0.05)。与HFSCs细胞组比较,lncRNA01535 mimics组细胞lncRNA01535 mRNA,JAK、STAT3 mRNA与蛋白表达水平升高(P<0.05),lncRNA01535 inhibitor组细胞lncRNA01535 mRNA,JAK、STAT3 mRNA与蛋白表达水平降低(P<0.05);与lncRNA01535 mimics组比较,lncRNA01535 inhibitor组细胞lncRNA01535 mRNA,JAK、STAT3 mRNA与蛋白表达水平降低(P<0.05)。结论lncRNA01535促进HFSCs的增殖和分化,其机制可能与lncRNA01535激活HFSCs中JAK/STAT3通路有关。

Objective To investigate the effect of LncRNA01535 on regulating the proliferation and differentiation of hair follicle stem cells(HFSCs)through Janus kinase/signal transduction and transcriptional activator 3(JAK/STAT3)signaling pathway.Methods The HFSCs cell group,LncRNA01535 overexpression group(LncRNA01535 mimics group),LncRNA01535 low expression group(LncRNA01535 inhibitor group)were set up.After 72 hours of culture,the level of cell survival was determined by CCK-8 assay,cell differentiation level was determined by red oil O staining,cell apoptosis and cell cycle were determined by flow cytometry and the real-time PCR(RT-PCR)and Western blot were used to determine the expression levels of LncRNA01535,JAK and STAT3 mRNA and protein.Results Compared with the HFSCs cell group,survival rate,and differentiation rate of the LncRNA01535 mimics group were increased,while the apoptosis rate and proportion of cells in the G_(1) phase were decreased(P<0.05).The survival rate and differentiation rate of the LncRNA01535 inhibitor group were decreased,whilethe apoptosis rate and proportion of G_(1) phase cells were increased(P<0.05).Compared with the LncRNA01535 mimics group,survival rate,and differentiation rate of the LncRNA01535 inhibitor group were decreased,while the apoptosis rate and proportion of G_(1) phase cells were increased(P<0.05).Compared with the HFSCs cell group,the expression levels of LncRNA01535 mRNA,JAK mRNA,STAT3 mRNA and protein in the LncRNA01535 mimics group were increased(P<0.05),and the expression levels of LncRNA01535 mRNA,JAK,STAT3 mRNA and protein in the LncRNA01535 inhibitor group were decreased(P<0.05).Compared with the LncRNA01535 mimics group,the expression levles of LncRNA01535 mRNA,JAK,STAT3 mRNA and protein of the LncRNA01535 inhibitor group were decreased(P<0.05).Conclusion LncRNA01535 can promote the proliferation and differentiation of HFSCs.The mechanism may be related to the activation of JAK/STAT3 signaling pathway in HFSCs by LncRNA01535.