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Korean Journal of Obstetrics & Gynecology 2005;48(7):1686-1697.
Published online July 1, 2005.
Proteomic analyses of cervical cancer tissues by two-dimensional gel electrophoresis and mass spectrometry.
Sung Ha Lee, Su Mi Bae, Ok Kyoung Kim, Hyun Jung Kim, Eun Kyung Park, Hae Nam Lee, Yong Wook Kim, Duck Yeong Ro, Joon Mo Lee, Sung Eun Namkoong, Young Lae Cho, Gye Hyun Nam, Byung Don Han, Yong Wan Kim, Chong Kook Kim, Woong Shick Ahn
1Department of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea. ahnws@catholic.ac.kr
2Cancer Research Center, Catholic Research Institutes of Medical Science, Korea.
3Department of Obstetrics and Gynecology, College of Medicine, Kyungpook National University, Korea.
4Department of Obstetrics and Gynecology, College of Medicine, Soonchunhyang University, Korea.
5KOMA Biotech, Korea.
6College of Pharmacy, Seoul National University, Korea.
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.
Key Words: Cervical cancer, Squamous cell carcinoma, Two-dimensional gel electrophoresis (2-DE), Matrix assisted laser desorption/ ionization-time of flighting (MALDI-TOF)
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