梅花鹿SCF基因参与色素合成的作用研究

doi:10.16431/j.cnki.1671-7236.2023.09.022

摘要/Abstract

摘要： 【目的】探究干细胞因子(stem cell factor,SCF)对梅花鹿色素合成过程的影响。【方法】构建梅花鹿SCF基因野生型(SCF1-9)和突变型(SCF789)表达载体并转染HEK293细胞。用孵育HEK293细胞的培养液培养小鼠黑色素瘤细胞(B16),模拟SCF在机体的转运过程,检测B16细胞的黑色素含量及酪氨酸酶活性;用RT-PCR法检测小眼症转录因子M型(MITF-M)、酪氨酸酶(TYR)、酪氨酸酶相关蛋白1(TYRP1)、TYRP2、溶质载体家族45成员2(SLC45A2)、SRY盒转录因子10(SOX10)和cAMP反应元件结合蛋白(CREB)的mRNA表达量;用Western boltting检测MITF-M、TYR蛋白表达水平。【结果】SCF1-9组黑色素含量、酪氨酸酶活性和MITF-M、TYR、SOX10基因mRNA表达量及TYR和MITF-M的蛋白表达量与对照组、SCF789组相比均极显著或显著升高(P<0.01;P<0.05)。SCF1-9组SCL45A2基因mRNA表达量与SCF789组相比显著升高(P<0.05),与对照组无显著差异(P＞0.05)。【结论】梅花鹿SCF基因通过SCF/c-kit信号通路调节色素基因MITF-M、SOX10、TYR、SLC45A2表达,促进色素合成。试验结果为进一步研究色素合成相关基因的表达调控提供理论依据。

关键词: 梅花鹿; SCF基因; 表达载体; 黑色素细胞

Abstract: 【Objective】 The purpose of this study was to explore the influence of stem cell factor (SCF) on the pigment synthesis process of sika deer.【Method】 Wild type (SCF1-9) and mutant (SCF789) expression vectors of sika deer SCF gene were constructed and transfected into HEK293 cells.Mouse melanoma cells (B16) were cultured with HEK293 cell culture medium to simulate the transport process of SCF in the body,and the melanin content and tyrosinase activity of B16 cells were detected.RT-PCR was used to detect the mRNA expression of microeye transcription factor M type (MITF-M),tyrosinase (TYR),tyrosinase-associated protein 1 (TYRP1),TYRP2,solute carry family 45 member 2 (SLC45A2),SRY box transcription factor 10 (SOX10) and cAMP response element-binding protein (CREB).The protein expression levels of MITF-M and TYR were detected by Western boltting.【Result】 Compared with control and SCF789 groups,the melanin content,tyrosinase activity,mRNA expression of MITF-M, TYR and SOX10 genes,and protein expression of TYR and MITF-M in SCF1-9 group were extremely significantly or significantly increased (P<0.01 or P<0.05).The mRNA expression of SCL45A2 gene in SCF1-9 group was significantly higher than that in SCF789 group (P<0.05),had no significant difference with control group (P>0.05).【Conclusion】 The SCF gene of sika deer regulated expression of pigment genes MITF-M,SOX10,TYR and SLC45A2 through SCF/c-kit signaling pathway to promote pigment synthesis.The results provided theoretical basis for further study on the expression regulation of pigment synthesis related genes.

Key words: sika deer; SCF gene; expression vector; melanocyte

中图分类号:

S825

陈旭, 董依萌, 邢秀梅, 杨苏坤, 刘欣. 梅花鹿SCF基因参与色素合成的作用研究[J]. 中国畜牧兽医, 2023, 50(9): 3663-3670.

CHEN Xu, DONG Yimeng, XING Xiumei, YANG Sukun, LIU Xin. Study on the Role of SCF Gene in Pigment Synthesis in Sika Deer[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3663-3670.

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