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Comparative Study
. 2004 Oct;130(10):615-22.
doi: 10.1007/s00432-004-0576-5. Epub 2004 Jul 9.

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

Jie-Feng Cui  1 , Yin-Kun Liu, Bo-Shen Pan, Hai-Yan Song, Yu Zhang, Rui-Xia Sun, Jie Chen, Ju-Tao Feng, Zhao-You Tang, Yan-Ling Yu, Hua-Li Shen, Peng-Yuan Yang
Affiliations
Comparative Study

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

Jie-Feng Cui et al. J Cancer Res Clin Oncol. 2004 Oct.

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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Figures

Fig. 1A–C
Fig. 1A–C
2-DE silver staining result of A Hep3B; B MHCC97L; C MHCC97H
Fig. 2A,B
Fig. 2A,B
Different protein spots in the CBB staining map of MHCC97H (A) and Hep3B (B) for ESI-MS/MS. The arrows represent different spots among HCC cell lines successfully identified by ESI-MS/MS
Fig. 3A–C
Fig. 3A–C
LC-ESI-MS/MS result for spot 12 (annexin 1). A spot 12 total ion chromatogram; B peptide MS map for spot 12 at 31 min; C tandem MS map for peptide1262
Fig. 4
Fig. 4
RT-PCR result for annexin1 and S100A4 in Hep3B, MHCC97L, and MHCC97H. A annexin1, S S100A4, C β-actin, 1 Hep3B, 2 MHCC97L, 3 MHCC97H, M 100 bp DNA ladder
Fig. 5A–F
Fig. 5A–F
The expression of annexin1 and S100A4 in HCC cell lines detected by immunohistochemistry (200×). Upper picture A MHCC97H; B MHCC97L; C Hep3B for annexin1. Bottom picture D MHCCC97H; E MHCC97L; F Hep3B for S100A4
See this image and copyright information in PMC

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