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Comparative proteomics and molecular mechanical analysis in CDA-II induced therapy of LCI-D20 hepatocellular carcinoma model

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Abstract

Purpose

To investigate the differential proteins and related molecular mechanism of CDA-II (cell differentiation agent-II) induced therapy on a human hepatocellular carcinoma model in nude mice with high metastatic potential (LCI-D20).

Methods

After tumors were transplanted 11 days, mice were intraperitoneally injected with CDA-II (1,800 mg/kg) for 20 days continuously. The tumor growth-inhibitory efficiency in CDA-II treated groups was calculated. Proteins extracted from tumor tissue were separated by two-dimensional gel electrophoresis (2DE) and the differential proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Western blotting (WB) was performed to verify the expression of certain candidate proteins. Reverse transcription-polymerase chain reaction (RT-PCR) was engaged to study the molecular mechanism of the therapy.

Results

CDA-II suppressed the growth and metastasis of tumor. The tumor growth-inhibitory efficiency was 41.8%. In total, 27 differentially expressed proteins were identified, including HSP27, UGDH, CK8, Hsp60, ENOA and AnxA5, with functions involved in oncogene expression and/or cell differentiation. In addition, apparent alternations of HSP60 and β-actin expression levels and their different posttranslational modifications (PTMs) were investigated. RT-PCR analysis confirmed that the cancer related genes c-myc, N-ras and MMP-9 were significantly down-regulated.

Conclusion

Our results demonstrate that CDA-II presence can change the proteome profiling and favors of the tumor suppression in LCI-D20 cell differentiation. Our results also suggest that the dynamic PTM of HSP60 expression levels could be used to predict HCC and might be a promising and useful biomarker to prognosticate CDA-II therapeutic efficacy.

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Abbreviations

CDA-II:

Cell differentiation agent II

HCC:

Hepatocellular carcinoma

LCI-D20:

A nude mice model for human HCC with high metastatic potential (Liver Cancer Institute, passage time 20 days)

2DE:

Two dimensional gel electrophoresis

MS:

Mass spectrometry

RT-PCR:

Reverse transcription-polymerase chain reaction

MALDI-TOF-MS:

Matrix assisted laser desorption/ionization-time of flight mass spectrometry

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Acknowledgments

This study was supported by National Natural Science Foundation of China (Grant No. 20675020), National 863 Program (2006AA02A308, 2007AA02Z479) and Shanghai Science and Technology Development Program (03DZ14024).

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Authors and Affiliations

  1. Chemistry Department of Fudan University, 220 Handan Rd, 200433, Shanghai, China

    Hui-zhi Fan, Hang Liu & Peng-yuan Yang

  2. Liver Cancer Institute, Zhongshan Hospital, Fudan University, 200032, Shanghai, China

    Hui-zhi Fan, Dong-mei Gao, Qun Xue, Jun Chen, Rui-xia Sun & Yin-kun Liu

  3. Institute of Biomedical Sciences, Fudan University, 200032, Shanghai, China

    Chen Zhang, Yin-kun Liu & Peng-yuan Yang

Authors
  1. Hui-zhi Fan
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  2. Hang Liu
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  3. Chen Zhang
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  4. Dong-mei Gao
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  5. Qun Xue
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  6. Jun Chen
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  8. Yin-kun Liu
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Correspondence to Yin-kun Liu or Peng-yuan Yang.

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Fan, Hz., Liu, H., Zhang, C. et al. Comparative proteomics and molecular mechanical analysis in CDA-II induced therapy of LCI-D20 hepatocellular carcinoma model. J Cancer Res Clin Oncol 135, 591–602 (2009). https://doi.org/10.1007/s00432-008-0493-0

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  • DOI: https://doi.org/10.1007/s00432-008-0493-0

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