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Cancer Proteomics : Hepatocellular Carcinoma

Cancer Proteomics : Hepatocellular Carcinoma. 생명과학부 임 승 외 2001.11.21. Content. Introduction(Cancer proteomics) Hepatocellular carcinomas(HCC) 2D-gel analysis 2D-gel electrophoresis Image analysis MALDI-TOF Identification Results. Genetic events in colorectal cancer. Introduction.

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Cancer Proteomics : Hepatocellular Carcinoma

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  1. Cancer Proteomics: Hepatocellular Carcinoma 생명과학부 임 승 외 2001.11.21

  2. Content • Introduction(Cancer proteomics) • Hepatocellular carcinomas(HCC) • 2D-gel analysis • 2D-gel electrophoresis • Image analysis • MALDI-TOF • Identification • Results

  3. Genetic events in colorectal cancer Introduction • Cancer proteomics : Protein-expression analysis -> Tumor marker or Drug target !

  4. 1. Sample Preparation 2.Separation 3. Analysis 4.Identification (of disease associated proteins) Normal Vs. Tumor 2D gels, HPLC Integration Mass spectrometry Bioinformatics Proteomics Research Flow Diagram

  5. Hepatocellular Carcinoma(HCC) • HCC 환자 중 90% 이상이 HBV, HCV에 의한(각각 70%, 20%) 만성 간질환을 동반 • Human의 경우, 대개 liver cirrhosis를 거쳐 HCC로 발전함. • 정상에서 HCC로 발전하기까지 약 20-40년이 소요. • Hepatocyte genome의 종양성 돌연변이

  6. Sample Preparation • Human cell line : HepG2, Huh7, Chang • Human tissue : Normal tissue Vs. Tumor tissue -> Total protein

  7. 2. Separation -1st IEF • IPG(immobilized pH gradients) strip -> Sample loading

  8. pH3 pH10 8.1 8.8 4.1 7.1 3.7 5.3 8.8 4.1 8.1 MW 7.1 5.3 3.7 2. Separation - 2nd SDS-PAGE • IEF가 끝난후 strip을 equiliburation 후 2nd SDS-PAGE로 Mw대로 분리. 2nd SDS-PAGE

  9. Gel Staining & Image Scan • CBB stain • Silver stain • SYPRO dye stain • RI labeling Human liver (SYPRO orange stain)

  10. 3. Analysis- Image Analysis Imagemaster 2D Elite 3.1

  11. 3. Analysis- MALDI-TOF • Matrix Assisted Laser Desorption Ionization - Time of Flight MALDI-TOF 동영상

  12. 4. Identification

  13. 5. Result 2001. Electrophoresis

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