卵泡刺激素对原癌基因c-myc在鸡卵泡上表达的影响  

Effects of follicle stimulating hormone on the expression of proto-oncogenes c-myc in chicken follicles

作　　者：刘小凡 谢晓磊 吴咏梅 陆应林[1] 虞德兵[1] LIU Xiaofan;XIE Xiaolei;WU Yongmei;LU Yinglin;YU Debing(College of Animal Science and Technology,Nanjing Agricultural university,Nanjing 210095,China;Laiyang Bureau of Agriculture and Rural Affairs of Shandong Province,Laiyang 265200,China)

机构地区：[1]南京农业大学动物科技学院,江苏南京210095 [2]山东省莱阳市农业农村局,山东莱阳265200

出　　处：《南京农业大学学报》2023年第3期564-571,共8页Journal of Nanjing Agricultural University

基　　金：江苏省农业科技自主创新资金项目[CX(17)3014]

摘　　要：[目的]本试验旨在研究c-myc基因在鸡卵泡上的表达及卵泡刺激素(FSH)对其的诱导作用,为明确c-myc在鸡卵泡发育过程中的相关机制提供理论基础。[方法]体外分离鸡小白卵泡(SWF)、大白卵泡(LWF)、小黄卵泡(SYF)、F4和F1卵泡颗粒层和膜层,检测c-myc mRNA和蛋白的表达;应用50 ng·mL^(-1)FSH处理小黄卵泡36 h,测量卵泡颗粒层的厚度和密度,检测颗粒层和膜层c-myc、标记基因和周期相关基因的表达;体外分离、培养小黄卵泡颗粒细胞,预培养16 h,用siRNA干扰c-myc 24 h后,加入50 ng·mL^(-1)FSH处理24 h,检测c-myc mRNA的表达。[结果]c-myc mRNA在等级前卵泡的颗粒层和膜层的表达水平都显著高于排卵前卵泡,卵泡的颗粒层和膜层上都有c-Myc蛋白表达。与对照组相比,FSH处理后,小黄卵泡颗粒层厚度增加约25.5%(P<0.05),密度增加约27.8%(P<0.01)。在小黄卵泡颗粒层上,FSH降低了c-myc和类固醇激素合成急性调节蛋白(star)mRNA的表达水平(P<0.05),提高了细胞增殖抗原(pcna)和周期蛋白D2(ccnd2)mRNA的表达水平(P<0.01);在卵泡膜层上,FSH提高了c-myc和pcna mRNA的表达水平(P<0.05),极显著提高了ccnd2和star mRNA的表达水平(P<0.01)。FSH能提高c-myc敲减后颗粒细胞c-myc mRNA的表达水平(P<0.05)。[结论]FSH能诱导c-myc在鸡颗粒细胞中的表达,推测c-myc与体外培养的鸡小黄卵泡颗粒细胞的终端分化有关。[Objectives]The aim of this study was to study the expression of c-myc in chicken follicles and the induction to c-myc by follicle stimulation hormone(FSH),and provide a theoretical basis for the study of the related mechanism of c-myc in chicken follicle development.[Methods]The granular and membrane layers of small white follicles(SWF),large white follicles(LWF),small yellow follicles(SYF),F4 and F1 follicles of chicken were isolated in vitro,and the expression of c-myc mRNA and protein was detected.SYF were treated with FSH(50 ng·mL^(-1))for 36 h in vitro,the thickness and density of the granular layer of follicles were measured,and the expression of c-myc,marker genes and cycle-related genes in the granular layer and membrane layer were detected.SYF granular cell(GC)were isolated and cultured 16 h in vitro,after small interference RNA(siRNA)was used to knockdown the expression of c-myc for 24 h,GC was treated with FSH(50 ng·mL^(-1))24 h,c-myc mRNA expression was detected.[Results]The expression level of c-myc mRNA in the granular layer and membrane layer of prehierarchical follicles was significantly higher than that of preovulation follicles,and c-Myc protein was expressed in the granular layer and membrane layer.Compared with the control group,FSH increased the thickness of granular layer of SYF by 25.5%(P<0.05)and the density by 27.8%(P<0.01).FSH decreased the gene expression level of c-myc and steroid hormone synthesis acute regulatory protein(StAR)(P<0.05),however increased the gene expression level of cell proliferation antigen(pcna)and cyclin D2(ccnd2)(P<0.01)on the granular layer of SYF;FSH increased the gene expression level of c-myc and pcna(P<0.05),and significantly increased the gene expression level of ccnd2 and star in follicular membrane layer(P<0.01).FSH could increase the c-myc gene expression level in GC after c-myc knockdown(P<0.05).[Conclusions]FSH can induce the expression of c-myc in chicken granular cells,and it is speculated that c-myc is related to terminal differentiation of chicke

关 键 词：鸡 C-MYC基因 卵泡刺激素(FSH) 卵泡 

分 类 号：S834[农业科学—畜牧学]