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논문KCI 등재

자궁경부암 조직에서 이차원 전기영동과 질량분석을이용한 단백질발현의 변화 분석

Proteomic analyses of cervical cancer tissues by two-dimensional gel electrophoresis and mass spectrometry

이준모|김용완|김종국|조영래|이해남|안웅식|김용욱|배수미|남계현|한병돈|남궁성은|김옥경|김현정|박은경|노덕영|이성하
대한산부인과학회 (Korean Soceity of Obstetrics and Gynecology)|2005.07|Obstetrics & Gynecology Science|제48권 제7호|pp. 1686-1697

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키워드

Cervical cancer질량분석Squamous cell carcinoma자궁경부편평상피세포암이차원 전기영동Two-dimensional gel electrophoresis (2-DE)Matrix assisted laser desorption/ ionization-time of flighting (MALDI-TOF)
주제분류산부인과학

초록

목적: 자궁경부 암조직에서 이차원 전기영동과 질량분석을 통한 단백질발현의 변화 분석. 연구방법: 정상 자궁경부 조직과 자궁 편평상피세포 암조직을 2-DE완충액을 사용하여 용해시킨 후, IEF를 실시하였다 (pH3-10 linear, IPG strip 17 cm). 그리고 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)를 실시하고 은 염색을 하였다. 스캔된 젤들은 PDQuest 2-D softwareTM을 사용하여 분석하였고, 변화가 확인된 단백질은 질량분석기를 이용하여 peptide의 스펙트럼을 얻은 후 Swiss-prot/TrEMBL와 Mascot의 데이터 정보 검색을 이용하여 단백질을 동정하였다. 결과: 본 연구에서 정상 자궁경부 조직과 자궁 편평상피세포 암조직에서 9개의 증가된 단백질과 (alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta-actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2), 7개의 감소된 단백질 (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) 그리고 6의 증가 또는 감소된 단백질 (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55)을 확인할 수 있었다. 결론: 자궁경부의 정상조직과 암조직 사이의 이차원 전기영동을 이용한 단백질 발현의 비교를 통하여 서로 다르게 발현되는 단백질 21개를 동정하였고, 이들 단백질 중 9개의 단백질은 기존에 다른 암 조직에서 암 표적물로 발표된 바 있으며, 나머지 12개의 단백질들은 자궁경부 편평 상피세포 암조직에 특이적인 종양표지물로 생각된다.

Abstract

Objective: Comparison of protein expressions by two-dimensional gel electrophoresis (2-DE) in normal cervix and squamous cell carcinoma tissues in Korean women. Methods: Normal cervix and squamous cell carcinoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH3-10 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sliver stain. Scanned image was analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrum identifications were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. Results: We found 9 up-regulation proteins (Alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta -actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2, unknown), 7 down-reguation proteins (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) and 6 up and down-regulation proteins (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55) between normal cervix and squamous cell carcinoma cell tissues.Conclusion: 2-DE offers total protein expressions between normal cervix and squamous cell carcinoma cell tissues, and searching of differently expressed protein for the diagnostic markers of squamous cell carcinoma tissue.

참고문헌 (39)

  1. 통계청, "Annual Report on the cause of death statistics 사망원인 통계연보 2001" (2001), 통계청
  2. Tommasino M, "The Role of TP53 in cervical carcinogenesis", 21(3), pp.307~12 (2003)
  3. Brenna SM, "Regulation of cell cycles is of key importance in human papillomavirus associated cervical carcinogenesis", 121(3), pp.128~32 (2003)
  4. Chen Y, "Identification of cervical cancer markers by cDNA and tissue microarrays", 63(8), pp.1927~35 (2003)
  5. Luo A, "Discovery of Ca -relevant and differentiation-associated genes downregulated in esophageal squamous cell carcinoma using cDNA microarray", pp.1~9 (2003)
  6. Cheng Q, "Identification of molecular markers for the early detection of human squamous cell carcinoma of the uterine cervix", 86(2), pp.274~81 (2002)
  7. Van Trappen PO, "A model for co-expression pattern analysis of genes implicated in angiogenesis and tumour cell invasion in cervical cancer", 87(5), pp.537~44 (2002)
  8. Alazawi W, "Changes in cervical keratinocyte gene expression associated with integration of human papillomavirus 16", 62(23), pp.6959~65 (2002)
  9. Cheng Q, "Identification and characterization of genes involved in the carcinogenesis of human squamous cell cervical carcinoma", 98(3), pp.419~26 (2002)
  10. Wilkins MR, "From proteins to proteomes:large-scale protein identification by two- dimensional electrophoresis and amino acid analysis", 14, pp.61~5 (1996)
  11. Mortz E, "Improved silver staining protocols for high sensitivity protein identification using matrix-assisted laser desorption", ionization-time of flight analysis, 2001, pp.1(11), 1359~63
  12. Humphery-Smith I, "Proteome research:complementarity and limitations with respect to the RNA and DNA worlds", 18, pp.1217~42 (1997)
  13. Pennington SR, "Proteome analysis:from protein characterization to biologycal function", 7, pp.168~73 (1997)
  14. Oh J, "Annexin-I inhibits PMA-induced c-fos SRE activation by suppressing cytosolic phospholipase A2 signal", 477(3), pp.244~8 (2000)
  15. Kim YS, "PKCdelta-dependent cleavage and nuclear translocation of annexin A1 by phorbol 12-myristate 13-acetate", 270(20), pp.4089~94 (2003)
  16. Alldridge LC, "The annexin protein lipocortin 1 regulates the MAPK", ERK pathway, 1999, pp.274(53), 37620~8
  17. Hannon R, "Aberrant inflammation and resistance to glucocorticoids in annexin 1-", - mouse, 2003, pp.17(2), 253~5
  18. Vercoutter-Edouart AS, "Proteomic analysis reveals that 14-3-3 sigma is down-regulated in human breast cancer cells", 61(1), pp.76~80 (2001)
  19. McConnell JE, "a kinase regulator whose expression pattern is modulated in mesenchyme and neuronal differentiation", 169(1), pp.218~28 (1995)
  20. Cell Signal 2000, "regulation of subcellular localization by molecular interference", 12, pp.703~70 (2000)
  21. Rosenquist M, "14-3-3 proteins in apoptosis", 36(4), pp.403~8 (2003)
  22. Solito E, "Dexamethasone induces rapid serine-phosphorylation and membrane translocation of annexin 1 in a human folliculostellate cell line via a novel nongenomic mechanism involving the glucocorticoid receptor", 144(4), pp.1164~74 (2003)
  23. Yaffe MB, "The structural basis for 14-3-3", 91(7), pp.961~71 (1997)
  24. Luk SC, "Developmental regulation of 14-3-3 epsilon isoform in rat heart", 68(2), pp.195~9 (1998)
  25. Kominea A, "Expression of the 27-kDa heat shock protein", 128(8), pp.426~32 (2002)
  26. Rane MJ, "Heat shock protein 27 controls apoptosis by regulating Akt activation", 278(30), pp.27828~35 (2003)
  27. Gunst SJ, "focal adhesion and small heat shock proteins in the regulation of actin remodeling and contractility in smooth muscle", 91(2), pp.963~72 (2001)
  28. Sasaki A, "Prognostic significance of heat shock protein 27 in patients with oral squamous cell carcinoma", 9(2), pp.341~4 (2002)
  29. Piotrowicz RS, "Heat shock protein 27 kDa expression and phosphorylation regulates endothelial cell migration", 12(14), pp.1481~90 (1998)
  30. Kaneko K, "Myosin light chain kinase inhibitors can block invasion and adhesion of human pancreatic cancer cell lines", 24(1), pp.34~41 (2002)
  31. Somlyo AP,, "Ca2+ sensitivity of smooth muscle and nonmuscle myosin II modulated by G proteins", 83(4), pp.1325~58 (2003)
  32. Von Bulow GU, "NF-AT activation induced by a CAML- interacting member of the tumor necrosis factor receptor superfamily", 278(5335), pp.138~41 (1997)
  33. Li L, "Evidence for serum response factor-mediated regulatory networks governing SM22alpha transcription in smooth", pp.311~21 (1997)
  34. Chang YS, "Enolase-alpha is frequently down-regulated in non-small cell lung cancer and predicts aggressive biological behavior", 9(101), pp.3641~4 (2003), Clin Cancer Res
  35. Arcuri D, "ENO1 gene product binds to the c-myc promoter and acts as a transcriptional repressor relationship with Myc romoter-binding protein 1", 473(1), pp.47~52 (2000)
  36. Banks-Schlegel SP, "Growth and differentiation of human esophageal carcinoma cell lines", 46(1), pp.250~8 (1986)
  37. Raval GN, "Loss of expression of tropomyosin-1 a novel class II tumor suppressor that induces anoikis", 22(40), pp.6194~203 (2003)
  38. , ~
  39. Schick C, "Squamous cell carcinoma antigen 2 is a novel serpin that inhibits the chymotrypsin-like proteinases cathepsin G and mast cell chymase", 272(3), pp.1849~55 (1997)

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