原癌基因c-fos在塔里木马鹿茸组织中表达特性的研究

摘要

为探讨原癌基因c-fos对鹿茸生长的调控作用,采用3头成年塔里木马鹿Cervus ela phus生长期为30、60d的新鲜鹿茸,剖分成茸皮层、间充质细胞层、成软骨细胞层和软骨细胞层.首先用2种免疫组化的方法进行基因表达定位,然后通过荧光定量PCR技术对不同组织基因的表达进行定量分析.免疫组化分析结果表明,该基因在茸皮的毛囊内根鞘和毛母质及皮脂腺、动脉血管的环形平滑肌处呈阳性反应；真皮乳头层与表皮基部连接的基底层呈阳性反应.在静脉血管、神经和其他附属器反应均未观察到阳性细胞.在间充质细胞层、成软骨层和软骨层2种方法均没有观察到c-fos的阳性表达细胞.定量分析发现,c-fos基因在不同生长阶段不同组织层均有表达,且在茸皮的表达量显著的高于间充质细胞层,成软骨层和软骨层(P＜0.05).在同一生长期间充质细胞层、成软骨层和软骨层c-fos的表达量很低；生长30与60 d比较,c-fos在间充质细胞层和成软骨层变化不大,在茸皮层和软骨层表现为下调表达.本研究表明c-fos基因在鹿茸快速生长期参与了茸皮干细胞的增殖与分化,并对成骨细胞的分化起着调控作用.%To study the effect of oncogene c-fos on velvet antler growth, antler tips were collected from 3 adult Tarim wapitis with 30- and 60-day-old growing antlers, respectively, and the samples were dissected into four zones, including epidermis/dermis, mesenchyme, precartilage and cartilage cell layers. Locations of c-fos expressions in the four zones of antler were determined by two immunohistochemical methods while quantitative analysis of c-fos expression was conducted with fluorescent quantitative PCR. The results demonstrated that the expression of c-fos was discovered in inner root sheath of hair follicle, hair matrix of dermal layer, and ring smooth muscle of artery while its weak expression was found in the basal cells between the papillary layer of der-mis and basal layers. There was no detectable expression in veins, nerve and other subsidiary organs, and the cells expressing c-fos were not detectable in mesenchyme, precartilage and cartilage cell layers by two methods. Analysis of quantitative PCR revealed that c-fos expressed in all layers at different growth stages. Higher expression level of c-fos was observed in dermal layer than that in mesenchymal cells layer, precartilage cells layer and cartilage(P<0. 05), And low expression of c-fos was found in mesenchymal cells layer, precartilage cells layer and cartilage. Little variation was found between the expressions of c-fos in mesenchymal and precartilage cells layer in the 30-and 60-day-old antlers while c-fos expression in epidermis and cartilage layers decreased with the increase of age. The results suggest that oncogene c-fos participated in the regulation of the proliferation and differentiation of stem cells in dermal layer during the rapid growth of the antlers, and c-fos may promote the differentiatiing of osteoblast into bone cells in cartilage cells layer.