MSX2基因在山羊真皮毛乳头细胞增殖中的调控作用

Regulatory Role of MSX2 in the Proliferation of Arbas White Cashmere Goat Dermal Papilla Cells

在山羊真皮毛乳头细胞中,建立MSX2基因过表达细胞株和MSX2基因干扰细胞株,通过生长曲线比较各细胞株的增殖能力,再通过Real-time PCR和Western Blot检测各株细胞中MSX2、LEF-1、cyclin D1基因的m RNA表达水平和蛋白质表达水平。结果表明,分离得到的山羊真皮毛乳头细胞均表达α-SMA和CD133表面标记,鉴定该细胞即为真皮毛乳头细胞。通过Real-time PCR检测各细胞株MSX2的表达量,发现在初级毛乳头细胞中过表达试验组是对照组的11.85倍,干扰试验组是对照组的0.31倍;而在次级毛乳头细胞中过表达试验组是对照组的10.59倍,干扰试验组是对照组的0.45倍,表明MSX2基因过表达和干扰细胞系已被成功建立。与此同时,研究中还发现,随着MSX2表达水平的增加,LEF-1和cyclin D1的表达水平也同步增加,同时使真皮毛乳头细胞增殖能力增强;随着MSX2表达水平的下调,LEF-1和cyclin D1的表达水平也同步减少,同时使真皮毛乳头细胞增殖能力下降。旨在探究MSX2基因在山羊真皮毛乳头细胞增殖中的调控模式,并为深入研究山羊毛囊的发生与生长提供理论基础。

MSX2 overexpression cell lines and MSX2 gene interference cell lines were established in goat dermal papilla cells. The growth curve was used to compare the proliferation ability of each cell line. Real-time PCR and Western Blot were used to detect MSX2 , LEF-1 , cyclin D1 gene mRNA expression levels and protein expression levels. The experimental results showed that the isolated goat dermal papilla cells express α-SMA and CD133 surface markers,and the cells were identified as dermal papilla cells. The expression of MSX2 was detected by Real-time PCR. In the primary dermal papilla cells overexpressing the experimental group was 11.85 times more than the control group,while the interfering group was 0.31 times that of the control group. In secondary dermal papilla cells,the overexpression of the experimental group was 10.59 times more than the control group,while the interfering group was 0.45 times that of the control group. This indicated that the MSX2 gene overexpression and interfering cell lines had been successfully established. At the same time,this study showed that with the increased of MSX2 expression,the expression of LEF-1 and cyclin D1 also increased synchronously,and at the same time triggered the dermal papilla cell proliferation. With the down-regulation of MSX2 expression,the expression levels of LEF-1 and cyclin D1 also decreased synchronously,at the same time,the proliferation of dermal dermal papilla cells decreased. The aim of this study was to investigate the regulatory pattern of MSX2 gene in the proliferation of goat dermal papilla cells. At the same time,it also will provides a theoretical basis for further study of the occurrence and growth of goat hair follicles.