苏博美利奴羊毛囊发育不同时期皮肤组织LncRNAs及其靶基因的差异表达
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摘要
长链非编码RNA (long noncoding RNA, LncRNA)参与哺乳动物多种生物学过程,目前研究其在毛囊形成过程中的作用机制却相对较少,因此研究LncRNA对毛囊发育的影响成为了解毛囊形成生物学过程的新方向。为验证前期获得的苏博美利奴羊(Ovis aries)毛囊发育不同时期皮肤组织的RNA-Seq结果以及与预测靶基因表达量差异,本研究利用qRT-PCR技术检测了在苏博美利奴羊毛囊发育6个关键时期两种在皮肤组织中差异表达且靶基因与毛囊发育相关的LncRNA 00202353和LncRNA 00316173及其靶基因的表达情况。结果表明,LncRNA 00202353及其靶基因分泌型卷曲相关蛋白2 (secreted frizzledrelated protein 2, SFRP2)的表达量在胚胎期85 d极显著高于胚胎期65 d、胚胎期135 d、出生后7 d和出生后30 d的表达量(P<0.01);LncRNA 00316173及其靶基因胰岛素样生长因子2 (insulin-like growth factor2, IGF2)的表达量在胚胎期65 d时极显著高于胚胎期135 d、出生后7 d和出生后30 d的表达量(P<0.01);将苏博美利奴羊毛囊发育时期分为发育期和成熟期进行分析,发现LncRNA 00202353及其靶基因SFRP2与LncRNA 00316173及其靶基因IGF2的表达量在毛囊发育期极显著高于毛囊成熟期(P<0.01),研究表明LncRNA 00202353与LncRNA 00316173及其靶基因参与毛囊的发育过程。本研究结果对后期深入研究LncRNA对毛囊发育的研究具有重要的参考价值,也为探究超细毛羊毛囊周期性变化的生物学过程提供研究线索。
Long noncoding RNA(LncRNA) is involved in a variety of biological processes in mammals, andthe current research on its role in hair follicle formation process is relatively small, therefore, studying the effect of LncRNA on hair follicle development has become a new direction to understand the biological process of hair follicle formation. In order to verify the RNA-Seq results of skin tissue in the non-period of the development of the Subo Merino sheep(Ovis aries) and the difference in the expression of predicted target genes, the expression of two LncRNA 00202353 and LncRNA 00316173 and their target genes in skin tissue,which were differentially expressed in skin tissue during the development of Subo Merino sheep and their target genes associated with hair follicle development were detected by qRT-PCR. The results showed that the expression level of LncRNA 00202353 and its target gene secreted frizzled-related protein 2(SFRP2) at 85 day of embryonic stage was extremely significantly higher than 65 and 135 d of embryonic stage, 7 and 30 d after birth(P<0.01). The expression level of LncRNA 00316173 and its target gene insulin-like growth factor2(IGF2) at 65 d of embryonic stage was extremely significantly higher than 135 d of embryonic stage, 7 and30 d after birth(P<0.01). Analysis of the hair follicle developmental stage of the Subo Merino sheep was divided into developmental and maturity stages, the expression levels of LncRNA 00202353 and its target gene SFRP2, LncRNA 00316173 and its target gene IGF2 were extremely significantly higher in the hair follicle development than in the hair follicle maturation(P<0.01). It indicatd that LncRNA 00202353 and LncRNA 00316173 and their target genes were involved in the development of hair follicles. These resullts provide important reference value for further studies on the development of hair follicles in LncRNAs, and supply research clues for exploring the biological processes of the periodic changes of ultrafine woolen sacs.
Long noncoding RNA(LncRNA) is involved in a variety of biological processes in mammals, andthe current research on its role in hair follicle formation process is relatively small, therefore, studying the effect of LncRNA on hair follicle development has become a new direction to understand the biological process of hair follicle formation. In order to verify the RNA-Seq results of skin tissue in the non-period of the development of the Subo Merino sheep(Ovis aries) and the difference in the expression of predicted target genes, the expression of two LncRNA 00202353 and LncRNA 00316173 and their target genes in skin tissue,which were differentially expressed in skin tissue during the development of Subo Merino sheep and their target genes associated with hair follicle development were detected by qRT-PCR. The results showed that the expression level of LncRNA 00202353 and its target gene secreted frizzled-related protein 2(SFRP2) at 85 day of embryonic stage was extremely significantly higher than 65 and 135 d of embryonic stage, 7 and 30 d after birth(P<0.01). The expression level of LncRNA 00316173 and its target gene insulin-like growth factor2(IGF2) at 65 d of embryonic stage was extremely significantly higher than 135 d of embryonic stage, 7 and30 d after birth(P<0.01). Analysis of the hair follicle developmental stage of the Subo Merino sheep was divided into developmental and maturity stages, the expression levels of LncRNA 00202353 and its target gene SFRP2, LncRNA 00316173 and its target gene IGF2 were extremely significantly higher in the hair follicle development than in the hair follicle maturation(P<0.01). It indicatd that LncRNA 00202353 and LncRNA 00316173 and their target genes were involved in the development of hair follicles. These resullts provide important reference value for further studies on the development of hair follicles in LncRNAs, and supply research clues for exploring the biological processes of the periodic changes of ultrafine woolen sacs.
引文
阿布来提·苏来曼,田月珍,许琴,等.2017.苏博美利奴羊胎儿皮肤毛囊结构及形态发育[J].中国农业科学,50(16):3226-3235.(Ablat Sulayman,Tian Y Z,Xu Q,et al.2017.Study on fetal skin hair follicle structure and morphological development of Subo Merino sheep[J].Scientia Agricultura Sinica,50(16):3226-3235.)
丁健华,蔡刚.2003.H19/IGF2基因印记调控机制研究进展[J].医学分子生物学杂志,25(4):241-245.(Ding J H,Cai G.2003.Research Progress of H19/IGF2 gene imprinting regulatory mechanism[J].Journal of Medical Molecular Biology,25(4):241-245.)
江玮,范一星,乔贤,等.2015.皮肤毛囊发育的转录组研究进展[J].遗传,37(6):528-534.(Jiang W,Fan Y X,Qiao X,et al.2015.The transcriptome research progresses of skin hair follicle development[J].Hereditas,37(6):528-534.)
李睿,罗云波.2016.lncRNA及其生物学功能[J].农业生物技术学报,24(4):600-612.(Li R,Luo Y B.2016.lncRNA and its biological function[J].Journal of Agricultural Biotechnology,24(4):600-612.)
邵勇,倪振洪,李玉红.2010.Wnt信号通路与毛囊干细胞[J].生物医学工程学杂志,27(4):946-948.(Shao Y,Ni Z H,Li Y H,et al.2010.Wnt signal transduction pathways and hair follicle stem cells[J].Journal of Biomedical Engineering,27(4):946-948.)
苏立宁,李华,旭日干,等.2011.毛囊分子调控机制[J].生命的化学,31(03):455-460.(Su L N,Li H,Xu R G,et al.2011.The research advance in the molecular regulation of hair follicles[J].Chemistry of Life,31(03):455-460.)
孙露露,石福岳,秦立志,等.2013.皖系长毛兔不同周龄Wnt10b、SFRP2基因在皮肤中的表达规律[J].遗传育种,49(13):4-8.(Sun L L,Shi F Y,Qin L Z,et al.2013.The expressions rules of Wnt10b and SFRP2 gene in skin of Wanxi angora rabbit[J].Genetics and Breeding,49(13):4-8.)
王宁,荣恩光,闫晓红.2012.毛囊发育与毛发生产研究进展[J].东北农业大学学报,43(9):6-11.(Wang N,Rong EG,Yan X H.Research progress of hair follicle development and hair production[J].Journal of Northeast Agricultural University,43(9):6-11.)
武晓英,任有蛇,张春香,等.2010.毛囊周期发育相关信号通路的研究进展[J].中国兽医杂志,46(6):69-71.(Wu X Y,Ren Y S,Zhang C X,et al.2010.Research progress of hair growth-related signaling pathway in hair follicles[J].Chinese Journal of Veterinary Medicine,46(6):69-71.)
柳楠,田可川,石国庆,等.2015.苏博美利奴羊核心群级进杂交不同世代对产毛性状影响的研究[J].中国畜牧杂志,51(15):6-10.(Liu N,Tian K C,Shi G Q,et al.Effect of different generations on wool traits during upgarding cross stages in Subo Merino nuleus herd[J].Chinese Journal of Animal Science,51(15):6-10.)
许永安,付小兵.2010.毛囊干细胞增殖与分化相关信号通路研究进展[J].中国修复重建外科杂志,24(02):161-164.(Xu Y A,Fu X B.2010.Development of hair follicle stem cells related signal transduction in proliferation and differentiation[J].Chinese Journal of Reparative and Reconstructive Surgery,24(02):161-164.)
杨峰,易凡,曹慧青,等.2014.长链非编码RNA研究进展[J].遗传,36(5):456-468.(Yang F,Yi F,Cao H Q,et al The emerging landscape of long non-coding RNAs[J]Hereditas,36(5):456-468.)
Arab K,Park Y J,Lindroth A M,et al.2014.Long noncoding RNA TARID directs demethylation and activation of the tumor suppressor TCF21 via GADD45A[J].Molecular Cell,55(4):604-614.
Chen L L,Carmichael G G.2010.Decoding the function of nuclear long non-coding RNAs[J].Current opinion in cell biology,22(3):357-364.
Higgins C A,Chen J C,Cerise J E,et al.2013.Microenvironmental reprogramming by three-dimensional culture enables dermal papilla cells to induce de novo human hairfollicle growth[J].roceedings of the National Academy of the Sciences of the USA,110(49):19679-19688.
Kim B K,Yoon S K.2014.Expression of sfrp2 is increased in catagen of hair follicles and inhibits keratinocyte proliferation[J].Annals of Dermatology,26(1):79-87.
Lei M,Schumacher L J,Lai Y C,et al.2017.Self-organization process in newborn skin organoid formation inspires strategy to restore hair regeneration of adult cells[J].Proceedings of the National Academy of the Sciences of the USA,114(34):e7101-e7110.
Lou X,Yuan H,Xie J,et al.2015.Growth factors have a protective effect on neomycin-induced hair cell loss[J].Cell Biology International,39(1):65-73.
Marchese F P,Raimondi I,Huarte M.2017.The multidimensional mechanisms of long noncoding RNA function[J].Genome Biology,18(1):206.
Mignone J L,Roig-Lopez J L,Fedtsova N,et al.2007.Neural potential of a stem cell population in the hair follicle[J].Cell Cycle,6(17):2161-2170.
Ren J,Jian F F,Jiang H,et al.2018.Decreased expression of SFRP2 promotes development of the pituitary corticotroph adenoma by upregulating Wnt signaling[J].International Journal of Oncology,52(6):1934-1946.
Wang S,Ge W,Luo Z,et al.2017.Integrated analysis of coding genes and non-coding RNAs during hair follicle cycle of cashmere goat(Capra hircus)[J].BMC Genomics,18(1):767.
Wu X,Brewer G.2012.The regulation of mRNA stability in mammalian cells:2.0[J].Gene,500(1):10-21.
Yue Y,Guo T,Yuan C,et al.2016.Integrated analysis of the roles of long noncoding RNA and coding RNA expression in sheep(Ovis aries)skin during initiation of secondary hair follicle[J].PLoS One,11(6):e0156890.
Zhou G,Kang D,Ma S,et al.2018.Integrative analysis reveals ncRNA-mediated molecular regulatory network driving secondary hair follicle regression in cashmere goats[J].BMC Genomics,19(1):222.
丁健华,蔡刚.2003.H19/IGF2基因印记调控机制研究进展[J].医学分子生物学杂志,25(4):241-245.(Ding J H,Cai G.2003.Research Progress of H19/IGF2 gene imprinting regulatory mechanism[J].Journal of Medical Molecular Biology,25(4):241-245.)
江玮,范一星,乔贤,等.2015.皮肤毛囊发育的转录组研究进展[J].遗传,37(6):528-534.(Jiang W,Fan Y X,Qiao X,et al.2015.The transcriptome research progresses of skin hair follicle development[J].Hereditas,37(6):528-534.)
李睿,罗云波.2016.lncRNA及其生物学功能[J].农业生物技术学报,24(4):600-612.(Li R,Luo Y B.2016.lncRNA and its biological function[J].Journal of Agricultural Biotechnology,24(4):600-612.)
邵勇,倪振洪,李玉红.2010.Wnt信号通路与毛囊干细胞[J].生物医学工程学杂志,27(4):946-948.(Shao Y,Ni Z H,Li Y H,et al.2010.Wnt signal transduction pathways and hair follicle stem cells[J].Journal of Biomedical Engineering,27(4):946-948.)
苏立宁,李华,旭日干,等.2011.毛囊分子调控机制[J].生命的化学,31(03):455-460.(Su L N,Li H,Xu R G,et al.2011.The research advance in the molecular regulation of hair follicles[J].Chemistry of Life,31(03):455-460.)
孙露露,石福岳,秦立志,等.2013.皖系长毛兔不同周龄Wnt10b、SFRP2基因在皮肤中的表达规律[J].遗传育种,49(13):4-8.(Sun L L,Shi F Y,Qin L Z,et al.2013.The expressions rules of Wnt10b and SFRP2 gene in skin of Wanxi angora rabbit[J].Genetics and Breeding,49(13):4-8.)
王宁,荣恩光,闫晓红.2012.毛囊发育与毛发生产研究进展[J].东北农业大学学报,43(9):6-11.(Wang N,Rong EG,Yan X H.Research progress of hair follicle development and hair production[J].Journal of Northeast Agricultural University,43(9):6-11.)
武晓英,任有蛇,张春香,等.2010.毛囊周期发育相关信号通路的研究进展[J].中国兽医杂志,46(6):69-71.(Wu X Y,Ren Y S,Zhang C X,et al.2010.Research progress of hair growth-related signaling pathway in hair follicles[J].Chinese Journal of Veterinary Medicine,46(6):69-71.)
柳楠,田可川,石国庆,等.2015.苏博美利奴羊核心群级进杂交不同世代对产毛性状影响的研究[J].中国畜牧杂志,51(15):6-10.(Liu N,Tian K C,Shi G Q,et al.Effect of different generations on wool traits during upgarding cross stages in Subo Merino nuleus herd[J].Chinese Journal of Animal Science,51(15):6-10.)
许永安,付小兵.2010.毛囊干细胞增殖与分化相关信号通路研究进展[J].中国修复重建外科杂志,24(02):161-164.(Xu Y A,Fu X B.2010.Development of hair follicle stem cells related signal transduction in proliferation and differentiation[J].Chinese Journal of Reparative and Reconstructive Surgery,24(02):161-164.)
杨峰,易凡,曹慧青,等.2014.长链非编码RNA研究进展[J].遗传,36(5):456-468.(Yang F,Yi F,Cao H Q,et al The emerging landscape of long non-coding RNAs[J]Hereditas,36(5):456-468.)
Arab K,Park Y J,Lindroth A M,et al.2014.Long noncoding RNA TARID directs demethylation and activation of the tumor suppressor TCF21 via GADD45A[J].Molecular Cell,55(4):604-614.
Chen L L,Carmichael G G.2010.Decoding the function of nuclear long non-coding RNAs[J].Current opinion in cell biology,22(3):357-364.
Higgins C A,Chen J C,Cerise J E,et al.2013.Microenvironmental reprogramming by three-dimensional culture enables dermal papilla cells to induce de novo human hairfollicle growth[J].roceedings of the National Academy of the Sciences of the USA,110(49):19679-19688.
Kim B K,Yoon S K.2014.Expression of sfrp2 is increased in catagen of hair follicles and inhibits keratinocyte proliferation[J].Annals of Dermatology,26(1):79-87.
Lei M,Schumacher L J,Lai Y C,et al.2017.Self-organization process in newborn skin organoid formation inspires strategy to restore hair regeneration of adult cells[J].Proceedings of the National Academy of the Sciences of the USA,114(34):e7101-e7110.
Lou X,Yuan H,Xie J,et al.2015.Growth factors have a protective effect on neomycin-induced hair cell loss[J].Cell Biology International,39(1):65-73.
Marchese F P,Raimondi I,Huarte M.2017.The multidimensional mechanisms of long noncoding RNA function[J].Genome Biology,18(1):206.
Mignone J L,Roig-Lopez J L,Fedtsova N,et al.2007.Neural potential of a stem cell population in the hair follicle[J].Cell Cycle,6(17):2161-2170.
Ren J,Jian F F,Jiang H,et al.2018.Decreased expression of SFRP2 promotes development of the pituitary corticotroph adenoma by upregulating Wnt signaling[J].International Journal of Oncology,52(6):1934-1946.
Wang S,Ge W,Luo Z,et al.2017.Integrated analysis of coding genes and non-coding RNAs during hair follicle cycle of cashmere goat(Capra hircus)[J].BMC Genomics,18(1):767.
Wu X,Brewer G.2012.The regulation of mRNA stability in mammalian cells:2.0[J].Gene,500(1):10-21.
Yue Y,Guo T,Yuan C,et al.2016.Integrated analysis of the roles of long noncoding RNA and coding RNA expression in sheep(Ovis aries)skin during initiation of secondary hair follicle[J].PLoS One,11(6):e0156890.
Zhou G,Kang D,Ma S,et al.2018.Integrative analysis reveals ncRNA-mediated molecular regulatory network driving secondary hair follicle regression in cashmere goats[J].BMC Genomics,19(1):222.