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Differential proteomic analysis of human hepatocellular carcinoma cell line metastasis-associated proteins

  • Original Paper
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Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

The comparative study of differentially expression of protein profiles of hepatocellular carcinoma cell lines with various metastasic potential and screening key molecules related to hepatocellular carcinoma metastasis and recurrence.

Methods

Using two-dimensional electrophoresis and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), we analyzed differentially displayed proteomics of human hepatocellular carcinoma cell lines Hep3B, MHCC97L, MHCC97H with different metastasic potential.

Results

Approximate 1,000 protein spots were detected on silver-stained gel by ImageMaster (977±113 spots in Hep3B, 1092±40 in MHCC97L, and 889±14 in MHCC97H). Fifty distinct different protein spots were analyzed with online LC-ESI-MS/MS. Only 26 protein spots had a positive result, including annexin1, S100A4, and so on. In comparison with nonmetastasis Hep3B cell lines, there were 16 proteins overexpressed in MHCC97H and MHCC97L, 10 proteins underexpressed in MHCC97H and MHCC97L. Applying cell immunohistochemistry and RT-PCR, we further validated two interesting and different proteins, annexin1 and S100A4.

Conclusion

The protein profile of metastatic hepatocellular carcinoma cell lines displayed obvious differences compared with non-metastatic liver cancer cell lines. The results imply that various different proteins may lead to HCC metastasis together.

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Fig. 1A–C
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Fig. 3A–C
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Fig. 5A–F

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Abbreviations

HCC:

Hepatocellular carcinoma

IPG-DALT:

Immobilized pH gradient-Dalton

2-DE:

Two-dimensional electrophoresis

LC-ESI-MS/MS:

Liquid chromatography-electrospray ionization-tandem mass spectrometry

RT-PCR:

Reverse transcription polymerase chain reaction

DMEM:

Dulbecco’s modified Eagle medium

CBB:

Coomassie brilliant blue

HPLC:

High performance liquid chromatography

PBS:

Phosphate buffered saline

BCIP/NBT:

5-bromo-4-chloro-3-indodyl phosphate /nitroblue tetrazolium

rARLP-1:

Rat aldose reductase-like protein1

Rak-C:

Rat aldo-keto reductase protein-C

References

  • Brunagel G, Schoen RE, Bauer AJ, Vietmeier BN, Getzenberg RH (2002) Nuclear matrix protein alterations associated with colon cancer metastasis to the liver. Clin Cancer Res 8:3039–3045

    CAS  PubMed  Google Scholar 

  • Coffey DS(2002) Nuclear matrix proteins as proteomic marks of preneoplastic and cancer lesions. Clin Cancer Res 8:3031–3033

    PubMed  Google Scholar 

  • Hanazaki K, Kajikawa S, Shimozawa N, Mihara M, Shimada K, Hiraguri M, Koide N, Adachi W, Amano J (2000) Survival and recurrence after hepatic resection of 386 consecutive patients with hepatocellular carcinoma. J Am Coll Surg 191:381–388

    Article  CAS  PubMed  Google Scholar 

  • Komatsu K, Kobune-Fujiwara Y, Andoh A, Ishiguro S, Hunai H, Suzuki N, Kameyama M, Murata K, Miyoshi J, Akedo H, Tatsuta M, Nakamura H (2000) Increased expression of S100A6 at the invading fronts of the primary lesion and liver metastasis in patients with colorectal adenocarcinoma. Br J Cancer 83:769–774

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Tang ZY, Ye SL, Liu YK, Chen J, Xue Q, Chen J, Gao DM, Bao WH (2001) Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97. World J Gastroenterol 7:630–636

    CAS  PubMed  Google Scholar 

  • Masaki T, Tokuda M, Ohnishi M, Watanabe S, Fujimura T, Miyamoto K, Itano T, Matsui H, Arima K, Shirai M, Maeba T, Sogawa K, Konishi R, Taniguchi K, Hatanaka Y, Hatase O, Nishioka M (1996) Enhanced expression of protein kinase substrate annexin1 in human hepatocellular carcinoma. Hepatology 24:72–81

    Article  CAS  PubMed  Google Scholar 

  • Nakamura T, Ajiki T, Murao S, Kamigaki T, Maeda S, Ku Y, Kuroda Y (2002) Prognostic significance of S100A4 expression in gallbladder cancer. Int J Oncol 20:937–941

    CAS  PubMed  Google Scholar 

  • Pedersen KB, Nesland JM, Fodstad, Malandsmo GM (2002) Expression of S100A4, E-Cadherin, α- and β-catenin in breast cancer biopsies. Br J Cancer 87:1281–1286

    Article  CAS  PubMed  Google Scholar 

  • Soskic V, Gorlach M, Poznanovic S, Boehmer FD, Godovac-Zimmermann J (1999) Functional proteomics analysis of signal transduction pathways of the platelet-derived growth factor βreceptor. Biochemistry 38:1757–1764

    Article  CAS  PubMed  Google Scholar 

  • Steiner S, Witzmann FA (2000) Proteomics: application and opportunities in preclinical drug development. Electrophoresis 21:2099–2104

    Article  CAS  PubMed  Google Scholar 

  • Tang ZY(2000) Surgery of hepatocellular carcinoma with special reference to studies on metastasis and recurrence. Gastroenterol Today 4:191–195

    Google Scholar 

  • Tang ZY(2003). Overview. In: Tang ZY (ed) Metastasis and recurrence of hepatocellular carcinoma basic and clinical studies. Shanghai Science and Technology, Shanghai, pp 1–24

  • Tang ZY, Yu YQ, Zhou XD, Ma ZC, Wu ZQ (1998) Progress and prospects in hepatocellular carcinoma surgery. Ann Chir 52:558–563

    CAS  PubMed  Google Scholar 

  • Tang ZY, Zhou XD, Li ZY, Yang BH, Ma ZC, Ye SL, Wu ZQ, Fan J, Liu YK, Liu KD, Qin LX, Tian J, Sun HC, He B, Xia JL, Qiu SJ, Zhou J (1999) Surgical treatment of hepatocellular carcinoma and related basic research with special reference to recurrence and metastasis. Chin Med J 112:887–891

    CAS  PubMed  Google Scholar 

  • Tian J, Tang ZY, Ye SL, Liu YK, Lin ZY, Chen J, Xue Q (1999) New human hepatocellular carcinoma (HCC) cell line with highly metastatic potential (MHCC97) and its expressions of the factors associated with metastasis. Br J Cancer 81:814–821

    Article  CAS  PubMed  Google Scholar 

  • Timperman AT, Aebersold R (2000) Peptide electroextraction for direct coupling of in gel digests with capillary LC-MS/MS for protein identification and sequencing. Anal Chem 72:4115–4121

    Article  CAS  PubMed  Google Scholar 

  • Wu WG, Tang XM, Hu W, Lotan R, Hong EK, Mao L (2002) Identification and validation of metastasis-associated protein in head and neck cancer cell line by two-dimensional electrophoresis and mass spectrometry. Clin Exp Metastasis 19:319–326

    Article  CAS  PubMed  Google Scholar 

  • Yu LR, Zeng R, Shao XX, Wang N, Xu YH, Xia QC (2000) Identification of differentially expressed proteins between human hepatoma and normal liver cell lines by two-dimensional electrophoresis and liquid chromatography-ion trap mass spectrometry. Electrophoresis 21:3058–3068

    Article  CAS  PubMed  Google Scholar 

  • Zeindl-Eberhart E, Klugbauer S, Dimitrijevic N, Jungblut PR, Lamer S, Rabes HM (2001) Proteome analysis of rat hepatomas: carcinogen-dependent tumor-associated protein variants. Electrophoresis 22:3009–3018

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This research was supported by the National Basic Research Priorities programme (001CB510205) and Nation Nature Science Foundation(30170416)

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Authors and Affiliations

  1. Liver Cancer Institute, Zhongshan Hospital, Fudan University, 136 Yi Xue Yuan Road, 200032, Shanghai, China

    Jie-Feng Cui, Yin-Kun Liu, Hai-Yan Song, Yu Zhang, Rui-Xia Sun, Jie Chen, Ju-Tao Feng & Zhao-You Tang

  2. Department of Clinical Laboratory Medcine, Zhongshan Hospital, Fudan University, Shanghai, China

    Jie-Feng Cui & Bo-Shen Pan

  3. Department of Chemistry, Fudan University, Shanghai, China

    Yan-Ling Yu, Hua-Li Shen & Peng-Yuan Yang

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  1. Jie-Feng Cui
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  2. Yin-Kun Liu
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  3. Bo-Shen Pan
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  4. Hai-Yan Song
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  5. Yu Zhang
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  7. Jie Chen
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  9. Zhao-You Tang
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  10. Yan-Ling Yu
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Corresponding author

Correspondence to Yin-Kun Liu.

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Cui, JF., Liu, YK., Pan, BS. et al. Differential proteomic analysis of human hepatocellular carcinoma cell line metastasis-associated proteins. J Cancer Res Clin Oncol 130, 615–622 (2004). https://doi.org/10.1007/s00432-004-0576-5

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  • DOI: https://doi.org/10.1007/s00432-004-0576-5

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