转录因子Foxq1通过WNT/β-catenin信号通路影响绒山羊毛囊干细胞增殖的研究  

Transcription Factor Foxq 1 Affects the Proliferation of Hair Follicle Stem Cells in Cashmere Goats via WNT/β-catenin Signaling Pathway

作　　者：许甜甜 张彤彤 王蒙 王昕[1] XU Tiantian;ZHANG Tongtong;WANG Meng;WANG Xin(College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100

出　　处：《畜牧兽医学报》2023年第6期2653-2661,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金面上项目(31972556)。

摘　　要：旨在探究陕北白绒山羊毛囊诱导期(E 66)和分化期(E 120)皮肤组织差异表达基因Foxq 1在绒山羊毛囊干细胞(HFSCs)中的功能。本研究选择年龄、体重基本一致的6只健康妊娠陕北白绒山羊母羊,在胚胎期E 66(n=3)和E 120(n=3)分别采集胎儿背部皮肤组织。将pAd-Foxq1和pAd-Blank过表达腺病毒分别侵染HFSCs,利用EdU、MTT及流式细胞等方法检测Foxq 1对HFSCs增殖的影响;利用qPCR和免疫荧光试验检测Wnt信号通路激活的标记因子CTNNB 1及Wnt信号通路下游基因的表达情况。qPCR结果表明,Foxq 1在绒山羊E 66和E 120皮肤组织差异表达(P<0.001),与测序数据一致;荧光显微镜下观察到彗星尾状荧光出现,表明Foxq 1腺病毒包装成功,且qPCR结果显示Foxq 1的过表达效率高达40000多倍(P<0.0001)。EdU和MTT结果表明,过表达Foxq 1后,毛囊干细胞的活力显著增强(P<0.05),EdU阳性细胞数量显著升高(P<0.01)。流式细胞周期结果显示,过表达Foxq 1后S期细胞比率显著升高。qPCR结果表明,过表达Foxq 1后CTNNB1、TCF 3及CYCLIND 1基因的mRNA表达量显著升高(P<0.05),免疫荧光进一步证明过表达Foxq 1后CTNNB1、TCF3及CYCLIND1蛋白水平的表达量显著升高。本研究发现,Foxq 1能够激活Wnt/β-catenin信号通路从而促进HFSCs的增殖,揭示了Foxq 1通过WNT/β-catenin信号通路影响绒山羊毛囊干细胞增殖的分子机理。This study aimed to explore the function of Foxq 1,a differentially expressed gene in the skin tissues at hair follicle induction stage(E 66)and cytodifferentiation stage(E 120),in hair follicle stem cells(HFSCs)of Shanbei white cashmere goat.In this study,6 healthy pregnant Shanbei white cashmere goats with the same age and body weight were selected.And the back skin tissues were collected at E 66(n=3)and E 120(n=3)from the fetuses,respectively.The pAd-Foxq1 and pAd-Blank overexpression adenovirus vectors were transfected into HFSCs.EdU,MTT and flow cytometry were used to detect the effects of Foxq 1 on the proliferation of HFSCs.qPCR and immunofluorescence assay were used to detect the expression of CTNNB 1,a marker of activating Wnt signaling pathway,and the downstream genes of Wnt signaling pathway.The results of qPCR showed that Foxq 1 was differentially expressed in the skin tissues of cashmere goats at E 66 and E 120 stages(P<0.001),which was consistent with the sequencing data.Comet tail fluorescence was observed under fluorescence microscope,indicating that Foxq 1 adenovirus was successfully packed,and qPCR results showed that the overexpression efficiency of Foxq 1 was more than 40000 times(P<0.0001).EdU and MTT results showed that the cell viability of hair follicle stem cells was significantly increased with the overexpression of Foxq1(P<0.05),and the number of EdU positive cells was significantly increased(P<0.01).Flow cytometry results showed that the percentage of S-phase cells was significantly increased with the overexpression of Foxq 1.qPCR results showed that the mRNA expressions of CTNNB1,TCF 3 and CYCLIND 1 genes were significantly increased with the overexpression of Foxq1(P<0.05).Immunofluorescence result further demonstrated that the protein expression levels of CTNNB1,TCF3 and CYCLIND1 were significantly increased with the overexpression of Foxq 1.This study revealed that Foxq 1 could activate the Wnt/β-catenin signaling pathway to promote the proliferation of HFSCs,which illustrated the

关 键 词：Foxq 1 毛囊干细胞 WNT信号通路 细胞增殖 绒山羊 

分 类 号：S827.2[农业科学—畜牧学]