사람 치수 세포와 치주 인대 세포의 유전자 발현에 관한 비교 연구
THE COMPARISON OF GENE EXPRESSION FROM HUMAN DENTAL PULP CELLS AND PERIODONTAL LIGAMENT CELLS
0
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
본 연구는 사람 치수세포 및 치주인대세포의 차이를 알아보고자 배양한 각각의 세포를 cDNA microarray assay를 통하여 유전자의 발현정도의 차이를 비교하였다. 그 결과를 바탕으로 각각의 세포에서 2배 이상의 유전자 발현의 차이를 보이는 유전자중 특징적인 3가지 유전자를 선택하여 RT-PCR로 검증한 결과 다음과 같은 결론을 얻었다; 1. Microarray assay 결과, 치주인대 세포에 비해 치수 세포에서 2배 이상 발현한 유전자 수는 총 51개가 나타났다. 2. RT-PCR의 결과, 치주인대세포에 비해 치수 세포에서 ITGA4, TGF-β2 등이 높게 나타났다. 3. Microarray assay 결과, 치수 세포에서 비해 치주인대 세포에서 2배 이상 발현한 유전자 수는 총 19개가 나타났다. 4. RT-PCR의 결과, 치수 세포에 비해 치주인대세포에서 LUM, WISP1, MMP1 등이 높게 나타났다. 본 연구 결과로 치수세포에는 상아질 형성에 관여하는 특징적인 유전자가 치주인대세포에 비해 높게 발현되었으며, 치주인대세포에는 교원질 합성에 관여하는 특징적인 유전자가 치수세포에 비해 높게 발현되어, 치수세포와 치주인대 세 포는 유전자 발현의 차이가 나타남을 알 수 있었다.
Abstract
The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA microarray assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well. According to this study, the results were as follows: 1. From the microarray assay, 51 genes were more expressed (2 fold) from PC than PDLC. 2. RT-PCR confirmed that ITGA4 and TGF β2 were more expressed in PC than in PDLC 3. From the microarray assay, 19 genes were more expressed (2 fold) from PDLC than PC. 4. RT-PCR confirmed that LUM, WISP1, and MMP1 were more expressed in PDLC than in PC. From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.
참고문헌 (38)
- Ruch JV, "Tooth morphogenesis and differentiation, in Dentin and Dentinogenesis", pp.47~79 (1984), CRC press
- Kikuchi H, "Effects of a functional agar surface on in vitro dentinogenesis induced in proteolytically isolated agar-coated dental papillae in rat mandibular incisors", Arch Oral Biol, 41, pp.871~883 (1996)
- Ruch JV, "Odontoblast differentiation and the formation of the odontoblast layer", J Dent Res, 64, pp.489~498 (1985)
- Osman M, "Behavior of odontoblast and basal lamina of trypsin or EDTA-isolated mouse dental papillae in short-term culture", J Dent Res, 60, pp.1015~1027 (1981)
- Ruch JV, "Odontoblast commitment and differentiation", Biochem cell Biol, 76, pp.923~938 (1998)
- Gronthos S, "Stem Cell Properties of Human Dental Pulp Stem Cells", J Dent Res, 81(8), pp.531~535 (2002)
- Gronthos S, "Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo", Proc Natl Acad Sci, 97, pp.13625~13630 (2000)
- Miura M, "SHED: Stem cells from human exfoliated decidous teeth", Proc Natl Acad Sci, 100, pp.5807~5812 (2003)
- McCulloch CA, "Origin and function of cells essential for periodontal repair: the role of fibroblasts in tissue homeostasis", Oral Dis, 1(4), pp.271~278 (1995)
- Seo BM, "Investigation of multipotent postnatal stem cells from human periodontal ligament", Lancet, 364, pp.149~155 (2004)
- P¨a¨akk¨onen V, "Analysis of gene and protein expression in healthy and carious tooth pulp with cDNA microarray and two-dimensional gel electrophoresis", Eur J Oral Sci, 113, pp.369~379 (2005)
- Shi S, "Comparison of Human Dental Pulp and Bone Marrow Stromal Cells by cDNA Microarray Analysis", Bone, 295, pp.532~539 (2001)
- Wang PL, "DNA microarray analysis of human gingival fibroblasts from healthy and inflammatory gingival tissues", Biochem Biophys Res Commun, 305, pp.970~973 (2003)
- Han X, "Identification of Genes Differentially Expressed in Cultured Human Osteoblasts Versus Human Fibroblasts by DNA Microarray Analysis", Connect Tissue Res, 43, pp.63~75 (2002)
- Nair PNR, "Pathogenesis of Apical Periodontitis and the Cause of Endodontic Failures", Crit Rev Oral Biol Med, 15(6), pp.348~381 (2004)
- Anand PS, "Management of Periodontitis Associated with Endodontically Involved Teeth: A Case Series", J Contemp Dent Pract, 2(6), pp.118~129 (2005)
- Satchell PG, "Apoptosis: an introduction for the endodontist", Int Endod J, 36, pp.237~245 (2003)
- Han X, "Identification of genes differentially expressed in cultured human periodontal ligament fibroblasts vs. human gingival fibroblasts by DNA microarray analysis", J Dent Res, 81(6), pp.399~405 (2002)
- Chen S, "Altered gene expression in human cleidocranial dysplasia dental pulp cells", Arch Oral Biol, 50, pp.227~236 (2005)
- Kamata N, "Immortalization of human dental papilla, dental pulp, periodontal ligament cells and gingival fibroblasts by telomerase reverse transcriptase", J Oral Pathol Med, 33, pp.417~423 (2004)
- Silva TA, "Dentin matrix proteins and soluble factors: intrinsic regulatory signals for healing and resorption of dental and periodontal tissues", Oral Diseases, 10, pp.63~74 (2004)
- Asano M, "Effect of connective tissue growth factor (CCN2", CTGF) on proliferation and differentiation of mouse periodontal ligament-derived cells, 2005, pp.5, 3~11
- Zhu Q, "Intergin expression in human dental pulp cells and their role in cell attachment on extracellular matrix proteins", J Endod, 24(10), pp.641~644 (1998)
- Luthman J, "Occurrence and distribution of different neurochemical markers in the human dental pulp", Arch Oral Biol, 37(3), pp.193~208 (1992)
- Beck LS, "In vivo induction of bone by recombinant human transforming growth factor beta 1", J Bone Miner Res, 6, pp.961~968 (1991)
- Machwate M, "Temporal variation of c-fos proto-oncogene expression during osteoblast differentiation and osteogenesis in developing rat bone", J Cell Biochem, 57, pp.62~70 (1995)
- Centrella M, "Transforming growth factor beta and remodeling of bone", J Bone Joint Surg Am, 73, pp.1418~1428 (1991)
- Ingram RT, "Effects of transforming growth factor beta and 1,25- dihydroxyvitamin D3 on the function, cytochemistry and morphology of normal human osteoblast-like cells", Differentiation, 55, pp.153~163 (1994)
- Ripamonti U, "Recombinant transforming growth factor beta induces endochondral bone in the baboon and synergizes with recombinant osteogenic protein-1(bone morphorgenetic protein-7) to initiate rapid bone formation", J Bone Miner Res, 12, pp.1584~1595 (1997)
- Rosen D, "Systemic adminstration of recombinant transforming growth factor beta 2 stimulates parameters of cancellous bone formation in juvenile and adult rats", Bone, 15, pp.355~359 (1994)
- Toyono T, "Expression of TGF-beta superfamily receptors in dental pulp", J Dent Res, 76, pp.1555~1560 (1997)
- Sloan AJ, "TGF-beta receptor expression in human odontoblasts and pulp cells", Histochem J, 31, pp.565~569 (1999)
- Gusman H, "Matrix metalloproteinase and gelatinolytic activity in clinically healthy and inflammed human dental pulps", Eur J Oral Sci, 110(5), pp.353~357 (2003)
- Shin SJ, "Tissue levels of matrix metalloproteinase in pulps and periapical lesions", J Endod, 28(4), pp.313~315 (2002)
- Lin SK, "Induction of dental pulp fibroblast matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase- 1 gene expression by interleukin-1 alpha and tumor necrosis factor alpha through a prostaglandin-dependent pathway", J Endod, 27(3), pp.185~189 (2001)
- Ababneh KT, "The proteoglycans of human cementum : immunohistochemcal localization in healthy, periodontally involved and ageing teeth", J Periodontal Res, 34(2), pp.87~96 (1999)
- Hall RC, "Immunochemical localization of the small leucin-rich proteoglycan lumican in human predentin and dentin", Arch Oral Biol, 42(10), pp.783~786 (1997)
- Chakravarti S, "Microarray studies reveal macrophage-like function of stromal keratocytes in the cornea", Invest Opthalmol Vis Sci, 45(10), pp.3475~3484 (2004)
