鸡VLDLR基因在不同组织中的表达分析
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摘要
极低密度脂蛋白受体(VLDLR)是低密度脂蛋白受体家族的一员,能以高亲和力结合富含Apo E的脂蛋白,是机体细胞摄取血液中富含甘油三酯的极低密度脂蛋白(VLDL)的主要受体。本研究旨在探讨VLDLR基因在鸡不同组织内的表达水平和分布情况,为进一步研究VLDLR在肿瘤发生中的变化提供依据。采集成年鸡骨骼肌、肺脏、脂肪、脾脏、肝脏等组织,提取RNA,根据Gen Bank中的鸡VLDLR基因序列设计引物并进行RT-PCR扩增,检测鸡5种组织中VLDLR基因的相对表达水平。结果显示VLDLR基因在鸡不同组织间的表达存在差异,表现为肌肉表达量最高,肺脏和脂肪组织次之,肝脏表达量最低。相对表达量由高到低依次为骨骼肌(10.95±0.14)、肺脏(3.87±0.23)、脂肪组织(3.02±0.11)、脾脏(2.31±0.08)和肝脏(1.00±0.17)。本试验证实VLDLR基因在肺脏有较高的表达量,脾脏也有一定的表达。
The very low density lipoprotein receptor( VLDLR) is a member of the low-density lipoprotein receptor family. It possesses high affinity in binding Apo E-rich lipoprotein and is the main receptor for uptake of blood triglyceride-rich very-low-density lipid protein( VLDL) in the body cells. This study was to investigate the expression level and distribution of the VLDLR gene in different tissues of chickens,and to provide evidence for the further study of VLDLR expression in tumorigenesis. Samples of lung,fat,spleen,liver and other tissues were collected quickly and stored in liquid nitrogen for RNA extraction. The expression of VLDLR in the five tissues of chickens was analyzed by designing primers based on chicken VLDLR sequence in Gen Bank and by performing RT-PCR amplification. The relative expression levels of VLDLR in the five tissues were detected. The results showed that the expressions of the VLDLR gene were different among the tissue samples: the highest in muscle,followed by lung and adipose tissue,and the lowest in liver. The relative expressions were that in skeletal muscle( 10. 95 ± 0. 14) > that in lung( 3. 87±0. 23) >that in adipose( 3. 02±0. 11) >that in spleen( 2. 31±0. 08) >that in liver( 1. 00±0. 17).To sum up,the VLDLR gene in the lungs had a higher expression,and that in the spleen had expression to a certain extent.
The very low density lipoprotein receptor( VLDLR) is a member of the low-density lipoprotein receptor family. It possesses high affinity in binding Apo E-rich lipoprotein and is the main receptor for uptake of blood triglyceride-rich very-low-density lipid protein( VLDL) in the body cells. This study was to investigate the expression level and distribution of the VLDLR gene in different tissues of chickens,and to provide evidence for the further study of VLDLR expression in tumorigenesis. Samples of lung,fat,spleen,liver and other tissues were collected quickly and stored in liquid nitrogen for RNA extraction. The expression of VLDLR in the five tissues of chickens was analyzed by designing primers based on chicken VLDLR sequence in Gen Bank and by performing RT-PCR amplification. The relative expression levels of VLDLR in the five tissues were detected. The results showed that the expressions of the VLDLR gene were different among the tissue samples: the highest in muscle,followed by lung and adipose tissue,and the lowest in liver. The relative expressions were that in skeletal muscle( 10. 95 ± 0. 14) > that in lung( 3. 87±0. 23) >that in adipose( 3. 02±0. 11) >that in spleen( 2. 31±0. 08) >that in liver( 1. 00±0. 17).To sum up,the VLDLR gene in the lungs had a higher expression,and that in the spleen had expression to a certain extent.
引文
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[13]杨璞.极低密度脂蛋白受体亚型在肿瘤细胞中的变化及其意义探讨[D].武汉:华中科技大学,2008.
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[15]Hussain MM.Structural,biochemi cal and signaling properties of the low-density lipoprotein receptor gene family[J].Front Biosci,2001(6):D417-428.
[16]刘志国,屈伸.极低密度脂蛋白受体研究进展[J].中国生物化学与分子生物学报,2003,19(6):683-689.
[17]Kreuter R,Souter A K,Wade D P,et al.Transcription factors CCAAT enhancer-binding proteinβand nuclear factor-Y bind to di screte regulatory elements in the very low density lipoprotein receptorpromoter[J].J Lipid Res,1999(40):376-386.
[2]Webb J C,Patel D D,Jones M D,et al.Characterization and tissue-specific expression of the human’very low density lipoprotein(VLDLR)receptor’m RNA[J].Hum Mol Genet,1994,3(4):531-537.
[3]Lijimah,Miyazawa M,Sakai J,et al.Expression and characterization of avery low density lipoprotein receptor variant lacking the OLink sugar regiongenerated by alternative splicing[J].J Biol Chem,1998,124:747-755.
[4]Tiebel O,Oka K,Robinson K,et al.Mouse very low-density lipoprotein receptor(VLDLR):Gene structure,tissue-specific expression and dietary and developmental regulation[J].Atherosclerosis,1999,145(2):239-251.
[5]苟华,胡深强.鸭极低密度脂蛋白受体和载脂蛋白E受体2在肌肉组织中的发育性表达模式研究[J].中国畜牧兽医,2014,41(3):7-13.
[6]Takahashi S,Sak AI J,Fujino T,et al.The very low density lipoprotein(VLDL)receptor-a peripheral lipoprotein receptor for remnant lipoproteins into fatty acid active tissues[J].Mol Cell Biochem,2003,248(1-2):121-127.
[7]Wyne K L,Patha R K,Seabra M C,et a1.Expression of the VLDL receptor in endothelial cells[J].Arteriosclerosis Thrombosis and Vascular Biology,1996,16(3):407-415.
[8]Takahashi S,Sak AI J,Fujino T,et al.The very low density lipoprotein(VLDL)receptor:characterization and functions as a peripheral lipoprotein receptor[J].J Ath Thro,2004,1(4):200-208.
[9]张金伟,张克英.产蛋鸡极低密度脂蛋白Y的转运与沉积机理[J].动物营养学报,2002,17(4):10-15.
[10]曹顶国,周艳,雷秋霞,等.鸡VLDLR基因多态性与蛋黄性状的关联分析[J].畜牧兽医学报,2012,43(6):849-856.
[11]Bujo H,Yamamoto T,Hayashi K,et al.Mutant oocytic low density lipoprotein receptor gene family member causes atherosclerosis and female sterility[J].Proc Natl Acad Sci,1995,92(21):9905-9909.
[12]何磊.Ⅱ型极低密度脂蛋白受体促肿瘤细胞侵袭迁移的作用及其机制研究[D].武汉:华中科技大学,2012.
[13]杨璞.极低密度脂蛋白受体亚型在肿瘤细胞中的变化及其意义探讨[D].武汉:华中科技大学,2008.
[14]Chen T,Wu F,Chen FM,et al.Variations of very low-density lipoprotein receptor subtype expression in gastrointestinal adenocarcinoma cells with various differentiations[J].World J Gastroenterol,2005,11(18):2817-2821.
[15]Hussain MM.Structural,biochemi cal and signaling properties of the low-density lipoprotein receptor gene family[J].Front Biosci,2001(6):D417-428.
[16]刘志国,屈伸.极低密度脂蛋白受体研究进展[J].中国生物化学与分子生物学报,2003,19(6):683-689.
[17]Kreuter R,Souter A K,Wade D P,et al.Transcription factors CCAAT enhancer-binding proteinβand nuclear factor-Y bind to di screte regulatory elements in the very low density lipoprotein receptorpromoter[J].J Lipid Res,1999(40):376-386.