사이클로포스파마이드에 인해 퇴축된 흰쥐 가슴샘의 재생과정에서 cDNA꼬리표를 이용한 발현 유전자의 특성 분석
Expression and Characterization of Genes by Expressed Sequence Tag Analysis in the Rat Thymus during Regeneration following Acute Thymic Involution Induced by Cyclophosphamide
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
가슴샘은 T 세포가 생성되는 면역계의 중추 기관이다. 가슴샘 퇴축으로 인해 감소된 면역력를 회복시키기 위해서는 가슴샘 재생과정의 분자기작을 이해하는 것이 필수적이다. 가슴샘의 재생과정에서 발현되는 유전자들을 분석하기 위해서, 재생중의 가슴샘 cDNA 라이브러리를 구축하였고, 무작위적으로 700개의 클론들을 선별하여 630개의 cDNA꼬리표를 확보하였다. 이를 분석한 결과 알려진유전자 486개(78%), 알려지지 않은 유전자 125개(19%) 그리고 새로운 유전자로 추정되는 cDNA꼬리표들을 포함하여 미확인유전자 19개(3%) 및 그 발현 빈도수를 확인하였다. 또한, 일반적인 기능별로 크게 6개의 범주로나누어 분류할 수 있었는데, 세포 대사에 관련된 유전자가 44%, 신호전달에 관련된 유전자가 20%, 막수송에 관련된 유전자가 7%, 그리고 세포골격, 세포분열 및 방어기전에 관련된 유전자가 각각 2%를 차지하였다. 확보된 cDNA꼬리표들 중 putative gene 01과 putative E2IG2 gene, musculin 및 osteoactivin 유전자의 발현이가슴샘 재생과정에서 현저하게 증가한다는 사실을 확인하였다. Putative gene 01의 유전자를 동정하기 위해 전체길이 cDNA의 염기서열을 분석하였고, 이에 대한 열린읽기틀을 확인하였다. 열린읽기틀 분석결과로 얻어진 아미노산 염기서열을 상동성 조사 및 아미노산 정렬을 실시한 결과 미토콘드리아 ribosomal protein S4와 아주 높은 상동성이 있음을 확인하였다. 따라서 본 연구의 결과는 가슴샘재생의 분자기전을 이해하는 데 유용한 정보를 제공할 뿐만 아니라 가슴샘의재생과 관련된 유용한 유전자를 발굴하는 데 기여할 수 있을 것이다.
Abstract
The thymus is the central lymphoid organ for the development of bone marrow-derived precursorcells into mature T-cells. Understanding the molecular mechanism of thymic involution and regeneration is criticalto develop methods to normalize or improve host immunity from the decreased immune function caused bythymic involution. In this study, the regenerating thymus cDNA library was constructed in the rat from a model of thymic involutionand regeneration induced by cyclophosphamide. Expressed sequence tags (ESTs) were obtained by partialsequencing of 700 randomly selected insert-containing clones. A total of 630 ESTs were analyzed, of which 486ESTs (78%) matched to known genes and 125 ESTs (19%) matched to other ESTs (unknown genes). The 19 ESTs(3%) did not match with any known sequences. The ESTs were grouped into six main functional categories: metabolism(44%), signaling components (20%), membrane transport (7%), cytoskeleton (2%), cell division (2%) anddefense (2%). As a result of RT-PCR analysis, expression of putative gene 01, putative E2IG2 gene, musculin and osteoactivinsignificantly increased in rat thymus during regeneration. The putative gene 01 showed complete homologywith mitochondrial ribosomal protein S4 by homology search and multiple alignment of amino acid. These results provide the extensive molecular information on thymus regeneration and will be useful source toidentify various genes which may play an important role in the thymus regeneration as well as to clone novel genes. Furthermore, the availability of these data will serve as a basis for further research to understand the molecularmechanism of thymus regeneration.
참고문헌 (30)
- Cunningham CP, "Thymic export in aged sheep: a continuous role for the thymus throughout pre- and postnatal life", Eur J Immunol, 31, pp.802~811 (2001)
- Mackall CL, "T-cell immunodeficiency following cytotoxic antineoplastic therapy: a review", Stem Cells, 181, pp.10~18 (2000)
- Yoon S, "Ultrastructural alterations of the cortical epithelial cells of the rat thymus after cyclophosphamide treatment", Histol Histopathol, 12, pp.401~413 (1997)
- Yoon S, "Upregulation of TrkA neurotrophin receptor expression in the thymic subcapsular, paraseptal, perivascular, and cortical epithelial cells during thymus regeneration", Histochem Cell Biol, 119, pp.55~68 (2003)
- Hinton PS, "IGF-I alters lymphocyte survival and regeneration in thymus and spleen after dexamethasone treatment", Am J Physiol, 274, pp.912~920 (1998)
- Awong G, "Key players for t-cell regeneration", Curr Opin Hematol, 17, pp.327~332 (2010)
- Olsen M, "A common language for physical mapping of the human genome", Science, 245, pp.1434~1435 (1989)
- Brown SM, "Sequence similarity searches on the world wide web", Biotechniques, 24, pp.248~250 (1998)
- Vander Heiden MG, "Bcl-xL prevents cell death following growth factor withdrawal by facilitating mitochondrial ATP", ADP exchange, 1999, pp.3, 159~167
- 김은희, "Adenine Nucleotide Translocator as a Regulator of Mitochondrial Function: Implication in the Pathogenesis of Metabolic Syndrome", Diabetes and Metabolism Journal, 34(3), pp.146~153 (2010), 대한당뇨병학회
- Gutiérrez-Aguilar M, "Physiological and pathological roles of mitochondrial SLC25 carriers", Biochem J, 454, pp.371~386 (2013)
- Jang JY, "Mitochondrial adenine nucleotide translocator 3 is regulated by IL-4 and IFN-gamma via STATdependent pathways", Cell Immunol, 226, pp.11~19 (2003)
- Smart N, "Thymosin beta4 and angiogenesis: modes of action and therapeutic potential", Angiogenesis, 10, pp.229~241 (2007)
- Sribenja S, "Roles and mechanisms of β-thymosins in cell migration and cancer metastasis: an update", Cancer Invest, 31, pp.103~110 (2013)
- Choi YL, "The primary structure of rat ribosomal protein L10: relationship to a Jun-binding protein and to a putative Wilms’ tumor suppressor", Biochem Biophys Res Commun, 225, pp.952~956 (1996)
- Hosoya K, "Unique behavior and function of the mitochondrial ribosomal protein S4(RPS4) in early dictyostelium development", Zoolog Sci, 12, pp.1455~1465 (2003)
- Probst P, "Identification and characterization of t cellstimulating antigens from Leishmania by CD4 t cell expression cloning", J Immunol, 166, pp.498~505 (2001)
- April HC, "Effects of estrogen on global gene expression:identification of novel targets of estrogen action", Cancer Res, 60, pp.5977~5983 (2000)
- Weintraub H, "The MyoD family and myogenesis: redundancy, networks, and thresholds", Cell, 75, pp.1241~1244 (1993)
- Porcher C, "The t cell leukemia oncoprotein SCL", tal-1 is essential for development of all hematopoietic lineages, 1996, pp.86, 47~57
- Bertrand N, "Proneural genes and the specification of neural cell types", Nat Rev Neurosci, 3, pp.517~530 (2002)
- Molkentin JD, "Defining the regulatory networks for muscle development", Curr Opin Genet Dev, 6, pp.445~453 (1996)
- Norton JD, "Id helix-loophelix proteins in cell growth and differentiation", Trends Cell Biol, 8, pp.58~65 (1998)
- Murre CC, "Structure and function of helix-loophelix proteins", Biochim Biophys Acta, 1218, pp.129~135 (1994)
- Robb L, "Musculin:a murine basic helix-loop-helix transcription factor gene expressed in embryonic skeletal muscle", Mechanisms of Development, 76, pp.197~201 (1998)
- Lu J, "Capsulin: a novel bHLH transcription factor expressed in epicardial progenitors and mesenchyme of visceral organs", Mechanisms of Development, 73, pp.23~32 (1998)
- Mendez-Ferrer S, "Mesenchymal and haematopoietic stem cells form a unique bone marrow niche", Nature, 466, pp.829~834 (2010)
- Singh M, "Functional roles of osteoactivin in normal and disease processes", Crit Rev Eukaryot Gene Expr, 20, pp.341~357 (2010)
- Sheng MH, "Osteoactivin is a novel osteoclastic protein and plays a key role in osteoclast differentiation and activity", FEBS Lett, 582, pp.1451~1458 (2008)
- Yoon TD, "Identification and analysis of expressed genes using a cDNA library from rat thymus during regeneration following cyclophosphamide-induced T cell depletion", Int J Mol Med, 31, pp.731~739 (2013)