不同年龄腰突出椎间盘组织CCNDBP1表达及意义
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摘要
目的比较细胞周期D1型结合蛋白1(CCNDBP1)在不同年龄阶段腰椎间盘突出症病人椎间盘中的表达,探讨其与腰椎间盘退变的关系。方法收集不同年龄阶段腰椎间盘突出症病人共42例,其中30～岁、40～岁、50～岁年龄段病人各14例。取病人突出的椎间盘组织,苏木精-伊红染色观察其退变情况,采用免疫组化法、PCR法、Western印迹法从细胞、基因、蛋白水平检测CCNDBP1的表达量。结果苏木精-伊红染色结果显示,各个年龄阶段腰椎间盘突出症病人腰椎间盘组织均有不同程度退变,退变程度与年龄呈正相关。免疫组化法、PCR法和Western印迹法检测结果显示,腰椎间盘组织CCNDBP1的表达随病人年龄的增加而减少,其中30～岁年龄段病人CCNDBP1的表达显著高于40～岁年龄段病人,差异有统计学意义(F=15.42～168.30,t=3.06～6.78,P<0.05),40～岁年龄段病人CCNDBP1的表达显著高于50～岁年龄段病人,差异有统计学意义(t=2.58～9.91,P<0.05)。结论人腰椎间盘髓核组织退变过程中有CCNDBP1的参与,随着年龄的增长,CCNDBP1在退变腰椎间盘组织中的表达量逐渐减少。
Objective To investigate the expression of cyclin-D1-binding protein 1(CCNDBP1)in the intervertebral disc of patients with lumbar disc herniation at different ages and its association with lumbar disc degeneration. Methods A total of 42 patients with lumbar disc herniation at different ages were enrolled,with 14 patients each in the 30-,40-,and 50-year groups.The prominent intervertebral disc tissue was collected,and HE staining was used to observe disc degeneration.Immunohistochemistry was used to measure the expression of CCNDBP1 in cells,and PCR and Western blot were used to measure the mRNA and protein expression of CCNDBP1. Results HE staining showed varying degrees of lumbar disc degeneration in the patients with lumbar disc herniation at different ages,and the degree of degeneration was positively correlated with age.Immunohistochemistry,PCR,and Western blot showed that the expression of CCNDBP1 in the intervertebral disc decreased with age;the 30-year group had significantly higher expression of CCNDBP1 than the 40-year group(F=15.42-168.30,t=3.06-6.78,P<0.05),and the40-year group had significantly higher expression of CCNDBP1 than the 50-year group(t=2.58-9.91,P<0.05). Conclusion CCNDBP1 is involved in the degeneration of human lumbar disc nucleus pulposus,and the expression of CCNDBP1 in the degenerated intervertebral disc gradually decreases with age.
Objective To investigate the expression of cyclin-D1-binding protein 1(CCNDBP1)in the intervertebral disc of patients with lumbar disc herniation at different ages and its association with lumbar disc degeneration. Methods A total of 42 patients with lumbar disc herniation at different ages were enrolled,with 14 patients each in the 30-,40-,and 50-year groups.The prominent intervertebral disc tissue was collected,and HE staining was used to observe disc degeneration.Immunohistochemistry was used to measure the expression of CCNDBP1 in cells,and PCR and Western blot were used to measure the mRNA and protein expression of CCNDBP1. Results HE staining showed varying degrees of lumbar disc degeneration in the patients with lumbar disc herniation at different ages,and the degree of degeneration was positively correlated with age.Immunohistochemistry,PCR,and Western blot showed that the expression of CCNDBP1 in the intervertebral disc decreased with age;the 30-year group had significantly higher expression of CCNDBP1 than the 40-year group(F=15.42-168.30,t=3.06-6.78,P<0.05),and the40-year group had significantly higher expression of CCNDBP1 than the 50-year group(t=2.58-9.91,P<0.05). Conclusion CCNDBP1 is involved in the degeneration of human lumbar disc nucleus pulposus,and the expression of CCNDBP1 in the degenerated intervertebral disc gradually decreases with age.
引文
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[14]KAWAGUCHI S,YAMASHITA T,YOKOGUSHI K,et al.Immunophenotypic analysis of the inflammatory infiltrates in herniated intervertebral discs[J].Spine,2001,26(11):1209-1214.
[15]TAPIAPEREZ H.Intervertebral disc pathologies from an immunological perspective[J].Revista de Neurologia,2008,46(12):751-757.
[16]TERAI S,AOKI H,ASHIDA K,et al.Human homologue of maid:a dominant inhibitory helix-loop-helix protein associated with liver-specific gene expression[J].Hepatology,2000,32(2):357-366.
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[18]XIA C,BAO Z,TABASSAM F,et al.GCIP,a novel human grap2and cyclin D interacting protein,regulates E2F-mediated transcriptional activity[J].Journal of Biological Chemistry,2000,275(27):20942-20948.
[19]BENEZRA R,DAVIS R L,LASSAR A,et al.The protein Id:a negative regulator of helix-loop-helix DNA binding proteins[J].Cell,1990,61(1):49-59.
[20]CHRISTY B A,SANDERS L K,LAU L F,et al.An Id-related helix-loop-helix protein encoded by agrowth factor-inducible gene[J].Proceedings of the National Academy of Sciences of the United States of America,1991,88(5):1815-1819.
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[24]MEZIANI R,YAMADA R,TAKAHASHI M,et al.A transethnic genetic study of rheumatoid arthritis identified FCGR2Aas a candidate common risk factor in Japanese and European populations[J].Japanese Journal of Rheumatology,2012,22(1):52-58.
[25]XIANG Hua,WANG Juan,MAO Yingwei,et al.Human telomerase accelerates growth of lens epithelial cells through regulation of the genes mediating RB/E2Fpathway[J].Oncogene,2002,21(23):3784-3791.
[26]MA W,STAFFORD L J,LI D,et al.GCIP/CCNDBP1,a helix-loop-helix protein,suppresses tumorigenesis[J].Journal of Cellular Biochemistry,2010,100(6):1376-1386.
[27]RUZINOVA,MARIANNA B,BENEZRA,et al.Id proteins in development,cell cycle and cancer[J].Trends in Cell Biology,2003,13(8):410-418.
[28]SIKDER H A,DEVLIN M K,DUNLAP S,et al.Id proteins in cell growth and tumorigenesis[J].Cancer Cell,2003,3(6):525-530.
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[30]NERLICH A G,WEILER C,ZIPPERER J,et al.Immunolocalization of phagocytic cells in normal and degenerated intervertebral discs[J].Spine,2003,28(15):2484-2490.
[2]殷相姣,宫赫,王丽珍,等.椎间盘退行性变的影响因素及其相关形态结构变化的研究进展[J].生物医学工程与临床,2016,33(1):112-117.
[3]曾佳兴,梁斌,尹东,等.青少年与中老年腰椎间盘突出相关因素分析[J].中国矫形外科杂志,2013,21(11):1121-1126.
[4]曾佳兴,梁斌,尹东,等.MMP-3、IgG和CD68在青少年与中老年突出腰椎间盘组织中的表达分析[J].中国脊柱脊髓杂志,2013,23(12):1109-1115.
[5]BABU N S,KRISHNAN S S,CHERUKUVADA V B,et al.Quantitative proteomic analysis of normal and degenerated human intervertebral disc[J].Spine Journal,2016,16(8):989-1000.
[6]王宗亮,刘雅,蔡明,等.组织蛋白酶K与人椎间盘退变相关性研究[J].创伤外科杂志,2009,11(3):245-247.
[7]MAITRE C L,FREEMONT A J,HOYLAND A A.The role of interleukin-1in the pathogenesis of human intervertebral disc degeneration[J].Arthritis Research&Therapy,2005,7(4):R732-R745.
[8]HARO H,KOMORI H,KATO T,et al.Experimental studies on the effects of recombinant human matrix metalloproteinases on herniated disc tissues-how to facilitate the natural resorption process of herniated discs[J].Journal of Orthopaedic Research,2010,23(2):412-419.
[9]ROBERTS S,EVANS H,MENAGE J,et al.TNFα-stimulated gene product(TSG-6)and its binding protein,IαI,in the human intervertebral disc:new molecules for the disc[J].European Spine Journal,2005,14(1):36-42.
[10]NAYLOR A,HAPPEY F,TURNER R L,et al.Enzymic and immunological activity in the intervertebral disk[J].The Orthopedic Clinics of North America,1975,6(1):51-58.
[11]HABTEMARIAM A,VIRRI J,GRONBLAD M,et al.The role of mast cells in disc herniation inflammation[J].Spine,1999,24(15):1516-1520.
[12]王沛,董强,雪原.腰椎间盘突出病变部的压力测定及其病理学意义[J].中华骨科杂志,2002,22(3):129-133.
[13]GEISS A,LARSSON K,RYDEVIK B,et al.Autoimmune properties of nucleus pulposus:an experimental study in pigs[J].Spine,2007,32(2):168-173.
[14]KAWAGUCHI S,YAMASHITA T,YOKOGUSHI K,et al.Immunophenotypic analysis of the inflammatory infiltrates in herniated intervertebral discs[J].Spine,2001,26(11):1209-1214.
[15]TAPIAPEREZ H.Intervertebral disc pathologies from an immunological perspective[J].Revista de Neurologia,2008,46(12):751-757.
[16]TERAI S,AOKI H,ASHIDA K,et al.Human homologue of maid:a dominant inhibitory helix-loop-helix protein associated with liver-specific gene expression[J].Hepatology,2000,32(2):357-366.
[17]YAO Y,DOKI Y W,IMOTO M,et al.Cloning and characterization of Dip1,a novel protein that is related to the Id family of proteins[J].Experimental Cell Research,2000,257(1):22-32.
[18]XIA C,BAO Z,TABASSAM F,et al.GCIP,a novel human grap2and cyclin D interacting protein,regulates E2F-mediated transcriptional activity[J].Journal of Biological Chemistry,2000,275(27):20942-20948.
[19]BENEZRA R,DAVIS R L,LASSAR A,et al.The protein Id:a negative regulator of helix-loop-helix DNA binding proteins[J].Cell,1990,61(1):49-59.
[20]CHRISTY B A,SANDERS L K,LAU L F,et al.An Id-related helix-loop-helix protein encoded by agrowth factor-inducible gene[J].Proceedings of the National Academy of Sciences of the United States of America,1991,88(5):1815-1819.
[21]DEED R W,BIANCHI S M,ATHERTON G T,et al.An immediate early human gene encodes an Id-like helix-loop-helix protein and is regulated by protein kinase C activation in diverse cell types[J].Oncogene,1993,8(3):599-607.
[22]RIECHMANN V,VAN C I,SABLITZKY F.The expression pattern of Id4,a novel dominant negative helix-loop-helix protein,is distinct from Id1,Id2and Id3[J].Nucleic Acids Research,1994,22(5):749-755.
[23]常献,陈斌,李长青.髓核细胞老化机制研究进展[J].中国矫形外科杂志,2014,22(1):40-42.
[24]MEZIANI R,YAMADA R,TAKAHASHI M,et al.A transethnic genetic study of rheumatoid arthritis identified FCGR2Aas a candidate common risk factor in Japanese and European populations[J].Japanese Journal of Rheumatology,2012,22(1):52-58.
[25]XIANG Hua,WANG Juan,MAO Yingwei,et al.Human telomerase accelerates growth of lens epithelial cells through regulation of the genes mediating RB/E2Fpathway[J].Oncogene,2002,21(23):3784-3791.
[26]MA W,STAFFORD L J,LI D,et al.GCIP/CCNDBP1,a helix-loop-helix protein,suppresses tumorigenesis[J].Journal of Cellular Biochemistry,2010,100(6):1376-1386.
[27]RUZINOVA,MARIANNA B,BENEZRA,et al.Id proteins in development,cell cycle and cancer[J].Trends in Cell Biology,2003,13(8):410-418.
[28]SIKDER H A,DEVLIN M K,DUNLAP S,et al.Id proteins in cell growth and tumorigenesis[J].Cancer Cell,2003,3(6):525-530.
[29]WANG Q,HII G,SHUSTERMAN S,et al.ID2expression is not associated with MYCN amplification or expression in human neuroblastomas[J].Cancer Research,2003,63(7):1631-1635.
[30]NERLICH A G,WEILER C,ZIPPERER J,et al.Immunolocalization of phagocytic cells in normal and degenerated intervertebral discs[J].Spine,2003,28(15):2484-2490.