家兔BMP2基因的克隆及其对毛乳头细胞增殖的影响  

Cloning of BMP2 gene in rabbits and its effect on the proliferation of hair papilla cells

作　　者：沈宁 李佳丽[1] 杨洁 赵博昊 陈阳[1] 吴信生[1] SHEN Ning;LI Jiali;YANG Jie;ZHAO Bohao;CHEN Yang;WU Xinsheng(College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《扬州大学学报(农业与生命科学版)》2024年第6期98-106,共9页Journal of Yangzhou University:Agricultural and Life Science Edition

基　　金：国家现代农业产业技术体系建设专项(CARS-43-A-1);国家自然科学基金资助项目(32102529、32072724)。

摘　　要：为探究家兔骨形成蛋白2(BMP2)基因对毛囊生长发育的影响,通过分子克隆获取家兔BMP2基因的编码序列,采用生物信息学对BMP2编码序列的生物学特性进行初步分析;并通过构建BMP2过表达载体和设计干扰序列,在毛乳头细胞(DPC)中过表达和敲低BMP2,利用qRT-PCR技术分析BMP2基因对毛囊发育相关基因(SFRP1、SFRP2、STAT1、CTNNB1、BCL2、HOXC13、EGF)mRNA表达的影响;通过CCK-8检测BMP2基因对DPC细胞增殖的影响。结果表明:家兔BMP2基因CDS全长为1188 bp,编码395个氨基酸;在DPC细胞中过表达BMP2后,SFRP1、SFRP2、STAT1基因表达量极显著提高,CTNNB1、BCL2、HOXC13基因表达量极显著下降,EGF基因表达量显著下降;siRNA-3敲低BMP2基因的表达后,SFRP1基因表达量显著下降,SFRP2、STAT1基因表达量极显著下降,CTNNB1、BCL2基因表达量极显著提高,HOXC13、EGF基因表达量显著提高。此外,BMP2能显著抑制DPC细胞增殖。综上,BMP2对动物毛囊发育具有重要的调控作用,能影响毛囊发育相关基因的表达,并抑制DPC细胞增殖。To investigate the effects of bone morphogenetic protein-2(BMP2)gene on hair follicle growth and development in rabbits,the coding sequence of the rabbit BMP2 gene was obtained by molecular cloning.Bioinformatic analysis was used to preliminarily analyze the biological characteristics of the BMP2 coding sequence.BMP2 overexpression and knockdown vectors were constructed,and designed interference sequences were used to overexpress and knockdown BMP2 in dermal papilla cells(DPCs).qRT-PCR was used to analyze the effects of BMP2 on the expression of hair follicle development-related genes(SFRP1,SFRP2,STAT1,CTNNB1,BCL2,HOXC13,EGF)mRNA.CCK-8 assay was used to detect the effects of BMP2 on DPC cell proliferation.The results showed that the full-length CDS of rabbit BMP2 gene was 1188 bp,encoding 395 amino acids.After overexpression of BMP2 in DPCs,the relative expression levels of SFRP1,SFRP2 and STAT1 genes were significantly increased(P<0.01),while the relative expression levels of CTNNB1,BCL2 and HOXC13 genes were significantly decreased(P<0.01),and the relative expression level of EGF gene was significantly decreased(P<0.05).After knockdown of BMP2 gene expression by siRNA-3,the relative expression level of SFRP1 gene was significantly decreased(P<0.05),the relative expression levels of SFRP2 and STAT1 genes were significantly decreased(P<0.01),the relative expression levels of CTNNB1 and BCL2 genes were significantly increased(P<0.01),and the relative expression levels of HOXC13 and EGF genes were significantly increased(P<0.05).Meanwhile,BMP2 significantly inhibited the proliferation of dermal papilla cells.BMP2 plays an important role in regulating animal hair follicle development,affecting the expression of hair follicle development-related genes and inhibiting the proliferation of dermal papilla cells.

关 键 词：家兔 骨形成蛋白2基因 毛乳头细胞 基因表达 

分 类 号：S858.291[农业科学—临床兽医学]