HOTAIR与山羊皮肤黑色素沉积的关系及体外表达规律的研究

doi:10.11843/j.issn.0366-6964.2020.03.007

摘要/Abstract

摘要： 旨在探讨山羊HOX转录反义RNA（HOTAIR）及其部分家族基因与山羊皮肤黑色素沉积的关系以及它们在黑色素细胞中的表达规律。本研究采用qRT-PCR检测HOTAIR和HOXD10、HOXC11、HOXC12基因在健康成年雌性酉州乌羊（（19.16±1.44）kg）、板角山羊（（23.27±3.24）kg）皮肤组织中的mRNA水平，及其在小鼠皮肤黑色素瘤细胞（B16-F10）增殖、分化过程中（培养1、3、5和7 d）的表达模式，并分析HOTAIR与其他3个基因表达的相关性。结果显示，在皮肤组织中，酉州乌羊HOTAIR和HOXC12的相对表达量极显著高于板角山羊（P<0.01），酉州乌羊HOXD10的相对表达量显著低于板角山羊（P<0.05）；HOTAIR与HOXC12表达显著正相关（P<0.05），与HOXD10表达显著负相关（P<0.05）。在B16-F10细胞增殖阶段，HOTAIR在各阶段表达差异不显著（P>0.05），HOXC11、HOXC12和HOXD10呈"高-低-高"的表达模式，且均在培养1 d的相对表达量极显著高于3和5 d（P<0.01）；HOTAIR与HOXC11表达呈极显著正相关（P<0.01）。在B16-F10细胞分化阶段，HOTAIR表达持续上升，HOXD10表达持续下降，HOXC11在各阶段的相对表达差异不显著（P>0.05），HOXC12在培养1和5 d的相对表达量极显著高于3和7 d（P<0.01）；HOTAIR与HOXD10表达极显著负相关（P<0.01）。HOTAIR在乌皮山羊皮肤中高表达，在B16-F10细胞分化阶段的表达随着培养时间的延长而增加，HOTAIR对HOXD10存在负向调控作用。HOTAIR与HOXD10互作可能调控山羊皮肤黑色素细胞的分化与黑色素生成。

关键词: 山羊, HOX转录反义RNA, HOX家族基因, 皮肤, 黑色素细胞

Abstract: The aim of this study was to explore the relationship between goat HOX transcript antisense RNA (HOTAIR) and its partial family genes and skin pigmentation in goats as well as its expression pattern in the melanocytes. qRT-PCR was used to detect the mRNA expression level of HOTAIR, HOXD10, HOXC11 and HOXC12 genes in skin tissues of adult female Youzhou Dark-skin goats ((19.16±1.44) kg) and Banjiao goats ((23.27±3.24) kg). Then the expression pattern of these genes in mouse skin melanoma cells (B16-F10) at different proliferation and differentiation stages (1, 3, 5 and 7 d cultured) were also detected by qRT-PCR, and the relationship in expression level between HOTAIR and the other 3 genes was analyzed. The results showed that expression of HOTAIR and HOXC12 in the skin tissue of Youzhou Dark-skin goats were significantly higher than those of Banjiao goats (P<0.01), however, expression of HOXD10 in the skin tissue of Youzhou Dark-skin goats was significantly lower than that of Banjiao goats (P<0.05). Pearson correlation analysis indicated that expression level of HOTAIR was significantly positively correlated with HOXC12 (P<0.05), but significantly negatively correlated with HOXD10 (P<0.05). In the proliferation stage of B16-F10 cells, expression of HOTAIR was not significantly different (P>0.05) among different proliferation stages; The expression of HOXC11, HOXC12 and HOXD10 showed a pattern of "high-low-high", and their mRNA levels on 1 d cultured were higher than those on 3 and 5 d cultured (P<0.01); The expression level of HOTAIR was positively correlated with HOXC11 (P<0.01). At the differentiation stage of B16-F10 cells, expression of HOTAIR kept increasing during cell differentiation; Instead, expression of HOXD10 kept decreasing during this stage; Expression of HOXC11 was not significantly different (P>0.05) among different differentiation stages; Expression of HOXC12 on 1 and 5 d cultured were very significantly higher than those on 3 and 7 d cultured (P<0.01); Expression level of HOTAIR was significantly negatively correlated with HOXD10 (P<0.01). HOTAIR shows high expression level in the dark skin of goats, and its expression level remains increasing during differentiation in B16-F10 cells. HOTAIR has a negative regulatory effect on HOXD10. The interaction between HOTAIR and HOXD10 may affect skin pigmentation by regulating the differentiation of skin melanocytes in goats.

Key words: goat, HOTAIR, HOX family gene, skin, melanocyte

中图分类号:

S827.2

蒋婧, 李杰, 周鹏, 王高富, 陈灿灿, 付琳, 张丽, 刘良佳, 任航行. HOTAIR与山羊皮肤黑色素沉积的关系及体外表达规律的研究[J]. 畜牧兽医学报, 2020, 51(3): 465-474.

JIANG Jing, LI Jie, ZHOU Peng, WANG Gaofu, CHEN Cancan, FU Lin, ZHANG Li, LIU Liangjia, REN Hangxing. The Relationship between HOTAIR and Skin Pigmentation in Goats and Its Expression Pattern in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(3): 465-474.

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