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- Title
Foxj2 基因对羊驼黑素细胞黑色素生成的影响.
- Authors
段志成; 董常生; 范瑞文; 谢建山; 杨玉静; 聂瑞强
- Abstract
Foxj2(Forkhead Box J2) gene, a member of the forkhead box transcription protein family, is positively correlated with the expression of epithelial-cadherin(E-cadherin, CDH1) in U87 cells and U251 cells, and CDH1 involved melanogenesis has a role in mediating the integrity of the epidermal-melanin unit. In order to verify that Foxj2 gene is involved in regulating melanogenesis in alpaca melanocytes, in this study, PCR method was used to verify the expression of Foxj2 gene in alpaca skin, the correlation between Foxj2 and melanogenesis related gene TYR, and Foxj2 and melanogenesis related proteins was analyzed by bioinformatics. Foxj2 siRNA and empty siRNA were synthesized, alpaca melanocyte model of silencing siRNA was constructed, and the transfection of cells was observed and recorded. Real-Time-PCR was used to detect the mRNA changes of Foxj2 gene and melanogenesis related gene TYR in the alpaca melanocytes of each group. Western blot was used to detect and compare the changes of Foxj2 and TYR protein levels in each group cell. Three kinds of melanin(total melanin, eumelanin, brown melanin) in the cells of each group were purified, and a microplate reader was used to measure the OD value of the three kinds of melanin in the cells of each group and compare changes. The results showed that Foxj2 gene expression was detected in alpaca melanocytes. Bioinformatics analysis showed that Foxj2 gene was involved in a series of functions such as amino acid transport regulation, growth factor and so on. The Forkhead domain existing in Foxj2 could have binding sites with the TYR gene promoter. Real-Time PCR results showed that compared with the NC group, the mRNA expression of Foxj2 in the silence group was extremely significantly decreased by 24 990%, and the expression of TYR was significantly increased by 35%. Western-blot results showed that the protein relative expression level of Foxj2 gene in the silence group extremely significantly decreased by 25%, and the protein relative expression level of TYR extremely significantly increased by 25%. The wavelength analysis of microplate reader showed that compared with the NC group, the total melanin relative expression levels of the silence group significantly increased by 88.0%, the eumelanin relative expression levels extremely significantly increased by 21.8%, and the pheomelanin relative expression level extremely significantly increased by 21.6%. Inhibiting the transcription of Foxj2 gene in alpaca melanocytes could increase melanogenesis in alpaca melanocytes, and play a role in promoting the main regulatory gene TYR of melanogenesis, proving that Foxj2 gene may play a regulatory role in melanogenesis in alpaca melanocytes.
- Subjects
GENE expression; BINDING sites; AMINO acid transport; MELANINS; FORKHEAD transcription factors; REGULATOR genes; MELANOGENESIS
- Publication
Journal of Shanxi Agricultural Sciences, 2023, Vol 51, Issue 6, p702
- ISSN
1002-2481
- Publication type
Article
- DOI
10.3969/j.issn.1002-2481.2023.06.14