단백질 질량분석과 바이오인포매틱스 기법을 이용한 암 특이적 단백질 마커의 발굴
Identification of protein biomarkers in endometrial cancer by using mass-spectrometry and bioinformatics
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프로테오믹스 기법과 바이오인포메틱스 기법을 융합하여 자궁내막암의 진단 및 치료의 타깃이 될 수 있는 단백질 바이오마커의 개발을 수행하였다. 인간 자궁내막암 세포주인 HEC-1A 세포에 발암과 암전이에 중요 인자로 알려진 신테칸-1을 감발현시키는 방법으로 암세포의 암거동을 변화시킨 후, 세포내에서 발현되는 단백질의 변화를 2-D 전기영동과 질량분석 방법으로 조사하였다. 대조군과의 비교를 통해 총 14가지의 단백질이 발현량에 차이가 나는 것으로 밝혀졌다. 이들의 펩티드 맵핑과 in silico DB 검색을 통해 각 단백질을 동정하였다. 이들은 기능에 따라 (i) 단백질의 안정성과 회전속도(turnover rate)에 관여하는 단백질 그룹, (ii) 산화스트레스에 반응하는 단백질 그룹, (iii) DNA 수선에 관여하는 단백질 그룹, (iv) RNA 처리와 운송(trafficking)에 관여하는 단백질 그룹으로 구분되었다. 장차 이들의 항체를 제작하여 자궁내막암의 진단 및 치료 수단으로서의 가능성을 열어놓았다.
Abstract
Syndecan-1 plays a key role in invasion and metastasis in endometrial cancer by helping liberate vital signals. Identification of proteins that associate with syndecan-1-enhanced cancer invasion and metastasis is a first step towards better response prediction and tailored treatment of patients. In the present study, we intended to identify putative protein biomarkers indicative of syndecan-1-induced invasion and metastasis in endometrial cancer, using 2-D gel electrophoresis coupled with MALDI-TOF mass-spectroscopy. Comparative proteome analysis was performed on human endometrial carcinoma (HEC)-1A cells that were treated with shRNA targeting syndecan-1, also referred to as 'HEC-1A/SDC-1 KD'. Peptides and proteins were identified by matching mass and pI values on 2-D gels obtained from HEC-1A/SDC-1A KD and its corresponding control cells by using an image enhancement software PDQuest. Comparative proteome analysis revealed 14putatively and differentially abundant proteins between HEC-1A and HEC-1A/SCD-1 KD cells, and identification of 14 differentially abundant proteins was performed by peptidemapping and the in silico database searching. They were further classified into subgroups based on their common functionality, including (i) protein stability and repair, (ii) oxidation stress, (iii) DNA repair (iv) RNA processing and trafficking. In the next studies, antibodies raised against these putative biomarkers will be tested for their relevances in endometrial cancer diagnosis and intervention.
참고문헌 (12)
- 주연, "프로테오믹스: 단백질에 대한 이해 및 기능 해석의 새로운 접근" (2004), 범문사
- Beauvais DM, "Syndecans in tumor cell adhesion and signaling", Reprod. Biol. Endocrino.l, 2, pp.1~12 (2004)
- Bernfield M, "Functions of cell surface heparan sulfate proteoglycans", Annu. Rev. Biochem, 68, pp.729~777 (1999)
- Bradford MM, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding", Anal. Biochem, 72, pp.248~254 (1976)
- 최동순, "Syndecan-1, a key regulator of cell viability in endometrial cancer", INTERNATIONAL JOURNAL OF CANCER, 121, pp.741~750, WILEY-LISS
- Gershenson DM, "Ovarian Cancer: Controversies in Management. 2nd Ed", pp.3~20 (1998), Churchill Livingstone Inc
- Kato M, "Loss of cell surface syndecan-1 causes epithelia to transform into anchorage-independent mesenchyme-like cells", Mol. Biol. Cell, 6, pp.559~576 (1995)
- Oakley BR, "A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels", Anal. Biochem, 105, pp.361~363 (1980)
- Rapraeger AC, "Molecular interactions of syndecans during development", Semin. Cell. Dev. Biol, 12, pp.107~116 (2001)
- Shevchenko A, "Mass spectrometric sequencing of proteins from silver-stained polyacrylamide", Gels. Anal. Chem, 68, pp.850~858 (1996)
- Westermeir R, "Proteomics in practice: a guide to successful experimental design. 3rd Ed", pp.35~70 (2006), Wley-VCH
- Zimmermann P, "The syndecans, tuners of transmembrane signaling", FASEB. J, 13, pp.S91~S100 (1999)