Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
권호 표지

Expression of amino acid transport system L in the differentiation of periodontal ligament fibroblast cells

치주인대섬유모세포의 분화과정에서 아미노산 수송계 L의 발현

  • Hwang, Kyu-Young (Department of Periodontology, College of Dentistry, Chosun University) ;
  • Kim, Do-Kyung (0ral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Kim, Chang-Hyun (0ral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Jang, Hyun-Seon (Department of Periodontology, Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Park, Joo-Cheol (0ral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Choi, Seong-Mi (Department of Periodontology, College of Dentistry, Chosun University) ;
  • Kim, Byung-Ock (Department of Periodontology, Oral Biology Research Institute, College of Dentistry, Chosun University)
  • 황규영 (조선대학교 치과대학 치주과학교실) ;
  • 김도경 (조선대학교 치과대학 구강생물학연구소) ;
  • 김창현 (조선대학교 치과대학 구강생물학연구소) ;
  • 장현선 (조선대학교 치과대학 치주과학교실, 구강생물학연구소) ;
  • 박주철 (조선대학교 치과대학 구강생물학연구소) ;
  • 최성미 (조선대학교 치과대학 치주과학교실) ;
  • 김병옥 (조선대학교 치과대학 치주과학교실, 구강생물학연구소)
  • Published : 2006.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

The periodontium is a topographically complex organ consisting of epithelial tissue, soft and mineralized tissues. Structures comprising the periodontium include the gingiva, periodontal ligament (PDL) , cementum and the alveolar bone. The molecular mechanism of differentiation in PDL fibroblast cells remain unclear. Amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In this study, the expression pattern of amino acid transport system L was, therefore, investigated in the differentiation of PDL fibroblast cells. To determine the expression level of amino acid transport system L participating in intracellular transport of amino acids in the differentiation of PDL fibroblast cells, it was examined by RT-PCR, observation of cell morphology, Alizaline red-S staining and uptake analysis after inducing experimental differentiation in PDL fibroblast cells isolated from mouse molar teeth. The results are as follows. 1. The LAT1 mRNA was expressed in the early stage of PDL fibroblast cell differentiation. This expression level was gradually reduced by differentiation- inducing time and it was not observed after the late stage. 2. The expression level of LAT2 mRNA was increased in time-dependent manner during differentiation induction of PDL fibroblast cells. 3. There was no changes in. the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during differentiation of PDL fibroblast cells. 4. The expression level of ALP mRNA was gradually increased and the expression level of Col I mRNA was decreased during differentiation of PDL fibroblast cells. 5. The L-leucine transport was reduced by time from the early stage to the late stage in PDL fibroblast cell differentiation. As the results, it is considered that among neutral ammo acid transport system L in differentiation of PDL fibroblast cells, the LATl has a key role in cell proliferation in the early stage of cell differentiation and the LAT2 has an important role in the late stage of cell differentiation for providing cells with neutral amino acids including several essential amino acids.

Keywords

References

  1. McCulloch CA, Lekic P, McKee MD. Role of physical forces in regulating the form and function of the periodontal ligament. Periodontol 2000 2000:24:56-72 https://doi.org/10.1034/j.1600-0757.2000.2240104.x
  2. Christensen HN. Role of amino acid transport and countertransport in nutrition and metabolism. Physiol Rev 1990:70:43-77 https://doi.org/10.1152/physrev.1990.70.1.43
  3. Kanai Y, Segawa H, Miyamoto K, et, al. Expression cloning and characterization of a transporter for large neutral amino acids activated by the heavy chain of 4F2 antigen (CD98). J Biol Chern 1998:273:23629-23632 https://doi.org/10.1074/jbc.273.37.23629
  4. Yanagida 0, Kanai Y, Chairoungdua A, et, al. Human L-type amino acid transporter 1 (LAT1): characterization of function and expression in turror cell lines. Biochim Biophys Acta 2001;1514:291-302 https://doi.org/10.1016/S0005-2736(01)00384-4
  5. Uchino H, Kanai Y, Kim DK, et, al. Transport of amino acid-related compounds mediated by L-type amino acid transporter 1 (LAT1): insights into the mechanism of substrate recognition. Mol Pharmacol 2002: 61:729-737 https://doi.org/10.1124/mol.61.4.729
  6. Nakamura E, Sato M, Yang H, et, al. 4F2 (CD98) heavy chain is associated covalently with an amino acid transporter and controls intracellular trafficking and membrane topology of 4F2 heterodimer. J Biol Chem 1999:274:3009-3016 https://doi.org/10.1074/jbc.274.5.3009
  7. Sang J, Lim YF, Panzica M, Finch P, Thompson NL. TAl, a highly conserved oncofetal complementary DNA from rat hepatoma, encodes an integral membrane protein associated with liver development, carcinogenesis, and cell activation. Cancer Res 1995:55:1152-1159
  8. Wolf DA, Wang S, Panzica MA, Bassily NH, Thompson NL. Expression of a highly conserved oncofetal gene, TAl/E16, in human colon carcinoma and other primary cancers: homology to Schistosoma mansoni amino acid permease and Caenorhabditis elegans gene products. Cancer Res 1996:56:5012-5022
  9. Segawa H, Fukasawa Y, Miyamoto K, et. al. Identification and functional characterization of a Na+-independent neutral amino acid transporter with broad substrate selectivity. J Biol Chem 1999:274:19745-19751 https://doi.org/10.1074/jbc.274.28.19745
  10. Verrey F, Meier C, Rossier G, Kuhn LC. Glycoprotein-associated amino acid exchangers: broadening the range of transport specificity. Pflugers Arch 2000:440:503-512 https://doi.org/10.1007/s004240000274
  11. Pineda M, Fernandez E, Torrents D, et, al, Identification of a membrane protein, LAT-2, that co-expressed with 4F2 heavy chain, an L-type amino acid transporter activity with broad specificity for small large zwitterionic amino aicds. J Biol Chem 1999:274:19738-19744 https://doi.org/10.1074/jbc.274.28.19738
  12. Rossier G, Meier C, Bauch C, et. al. LAT2, a new basolateral 4F2hc/CD98-associated amino acid transporter of kindney and intestine. J Biol Chem 1999:274:34948-34954 https://doi.org/10.1074/jbc.274.49.34948
  13. Shibuya N, Nemoto E, Kanaya S, Kunii R, Shimauchi H. Retinoic acid is a potential negative regulator for differentiation of human periodontal ligament cells. J Periodontal Res 2005;40:432-440 https://doi.org/10.1111/j.1600-0765.2005.00811.x
  14. Kim DK, Kanai Y, Choi HW, et, al, Characterization of the system L amino acid transporter in T24 human bladder carcinoma cells. Biochim Biophys Acta 2002;1565:112-121 https://doi.org/10.1016/S0005-2736(02)00516-3
  15. Yoon JH, Kim YE, Kim MS, et, al, Expression and functional characterization 794 of the system L amino acid transporter in KB human oral epidermoid carcinoma cells. Cancer Lett 2004;205:215-226 https://doi.org/10.1016/j.canlet.2003.10.009
  16. Foster BL, Nociti FH Jr, Swanson EC, et. al. Regulation of cementoblast gene expression by inorganic phosphate in vitro. Calcif Tissue Int 2006;78:103-112 https://doi.org/10.1007/s00223-005-0184-7
  17. Yoon JH, Kim lJ, Kim H, et. al. Amino acid transport system L is differently expressed in human normal oral keratinocytes and human oral cancer cells. Cancer Lett 2005; 222:237-245 https://doi.org/10.1016/j.canlet.2004.09.040