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정상 자궁 근육조직과 자궁근종에서의 이차원전기영동을 이용한 단백질 발현의 변화

Comparison of Protein Expression in Normal Myometrium and UterineLeiomyoma Using Two-Dimensional Gel Electrophoresis in Korean Women

박은경|김민숙|이준모|김용욱|안웅식|배수미|남궁성은|김고운|김종국|노덕영|이승구
대한산부인과학회 (Korean Soceity of Obstetrics and Gynecology)|2004.04|Obstetrics & Gynecology Science|제47권 제4호|pp. 618-626

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키워드

암Cancer자궁근종질량분석자궁경부이차원 전기영동Two-dimensional gel electrophoresis (2-DE)Leiomyoma. MyometriumMatrix assisted laser desorption/ionization-time of flighting (MALDI-TOF)
주제분류산부인과학

초록

목적 : 한국 여성의 자궁 근육조직과 자궁근종에서의 단백질 발현을 비교하기 위하여 이차원 전기영동을 실시하였다. 연구 방법 : 6개의 자궁 근육조직과 자궁근종을 이차원 전기 영동 용액에 용해하였고, IPG strip (17 cm pH 4-8)를 사용하여 isoelectric focusing을 실시한 후, 12% SDS-PAGE를 이용하여 단백질을 전개한 후, 은 염색을 하여 관찰하였고, PDQuest 2-D software를 이용하여 분석하였다. 단백질은 질량분석기를 이용하여 각각의 스펙트럼을 얻었고, Swiss-prot/TrEMBL와 Mascot 그리고 MS-FIT 데이터를 이용하여 단백질을 확인하였다. 결과 : 자궁 근육조직과 자궁근종의 단백질 발현 비교에서 발현이 증가하는 17개의 단백질 (phosphate carrier protein, 60 kDa heat shock protein, acidic calcium-independent, glutathione transferase omega, chloride intracellular channel 4, Ras-related protein Rab-11B, phosphatidylinositol transfer protein alpha isoform, type II keratin subunit protein, Cofilin 2 isoform 1, transgelin, ATP carrier protein, alpha-catenin homolog, parkinson disease 2, apo- cellular retinoic acid binding protein II, osteoglycin preproprotein, proteasome activator subunit 1 isoform, Unnamed protein)과 감소하는 7개의 단백질 (Serum amyloid P component, annexin IV, alpha 1 actin precursor, hypoxanthine-guanine phosphoribosyltransferase, tumor necrosis factor receptor superfamily member EDAR precursor, peroxiredoxin 2, translation elongation factor EF-Tu precursor)을 확인하였다. 결론 : 이차원 전기영동을 이용해서 자궁 근육조직과 자궁근종의 전체단백질 발현의 정도를 보여주었고, 두 조직간에 서로 다르게 발현되는 단백질을 확인하였다.

Abstract

Objective : Comparison of protein expression by two-dimensional gel electrophoresis (2-DE) in normal myometrium and uterine leiomyoma in Korean women. Methods : Normal myometrium and uterine leiomyoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH4-8 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) and sliver stain. Scanned image 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 spectrums identification were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. Results : In this study, we found 17 up-regulated proteins (phosphate carrier protein, 60 kDa heat shock protein, acidic calcium-independent, glutathione transferase omega, chloride intracellular channel 4, Ras-related protein Rab-11B, phosphatidylinositol transfer protein alpha isoform, type II keratin subunit protein, Cofilin 2 isoform 1, transgelin, ATP carrier protein, alpha-catenin homolog, parkinson disease 2, apo-cellular retinoic acid binding protein II, osteoglycin preproprotein, proteasome activator subunit 1 isoform, Unnamed protein) and 7 down-regulated proteins (Serum amyloid P component, annexin IV, alpha 1 actin precursor, hypoxanthine-guanine phosphoribosyltransferase, tumor necrosis factor receptor superfamily member EDAR precursor, peroxiredoxin 2, translation elongation factor EF-Tu precursor) between myometrium and leiomyoma. Conclusion : 2-DE offer total protein expression between normal myometrium and uterine leiomyoma, and searching of differently expressed protein for the diagnostic markers of leiomyoma.

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