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Identification of side population cells in human hepatocellular carcinoma cell lines with stepwise metastatic potentials

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Abstract

Purpose

To identify the side population (SP) cells from four hepatocellular carcinoma (HCC) cell lines with stepwise metastatic potentials.

Methods

SP cells were sorted from HCCLM3, MHCC97-H, MHCC97-L and Hep3B by flow cytometry, and then analyzed by differentiation study, clonogenic assay, chemoresistance study and tumorigenicity assay in vivo. The expression of ABCG2 in SP cells was detected by immunocytochemistry, western blotting and real-time quantitative PCR, respectively.

Results

There was significant difference in SP proportion among HCCLM3, MHCC97-H, MHCC97-L and Hep3B (28.7 ± 1.6%, 14.5 ± 0.6%, 4.2 ± 0.4%, 0.9 ± 0.1%, respectively, P < 0.01). All the SP cells showed similar characteristics of self-renewal, high clonogenicity, remarkable chemo-resistance and high expression of ABCG2. As low as 2,000 SP cells could initiate tumors in non-obese diabetic/severe combined immunodeficiency mice successfully.

Conclusions

SP cells purified from HCC cell lines harbors cancer stem cell-like properties, and may be related to the metastatic potentials and therapeutic-resistance of HCC.

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Abbreviations

HCC:

Hepatocellular carcinoma

SP:

Side population

ABCG2 :

ATP-binding cassette transporter family G2

BCRP1:

Breast cancer resistance protein1

MP:

Main population

CSCs:

Cancer stem cells

NOD/SCID:

Non-obese diabetic/severe combined immunodeficiency

FCM:

Flow cytometry

PI:

Propidium iodide

LIF:

Leukemia inhibitory factor

5-FU:

5-Fluorouracil

MMC:

Mitomycin C

ICC:

Immunocytochemistry

TACE:

Transcatheter arterial chemoembolization

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Acknowledgments

We thank Drs. Guo-ping Zhang for his expertise in flow cytometry. This study was combinedly supported by National Natural Science Foundation of China (30500594, 30571801); Shanghai Science and Technology Development Funds (06QA14012); Foundation of Shanghai Science Technology Commission (07JC14010, 06xD14004, 044119608); the Program for Excellent Disciplinary Leaders of Shanghai Health Bureau (LJ06004) and the Foundation of China National “211” Project for Higher Education. The funding agencies had no role in the design, conduct, decision to report or writing of the study.

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Authors and Affiliations

  1. Key Laboratory for Carcinogenesis and Cancer Invasion, Liver Cancer Institute, Zhongshan Hospital and Shanghai Medical School, Fudan University, the Chinese Ministry of Education, 136 Yi Xue Yuan Road, Shanghai, 200032, People’s Republic of China

    Guo-Ming Shi, Yang Xu, Jia Fan, Jian Zhou, Xin-Rong Yang, Shuang-Jian Qiu, Wei-Zhong Wu, Yuan Ji, Ai-Wu Ke, Zhen-Bin Ding, Yi-Zhou He, Bing Wu, Guo-Huan Yang, Wen-Zhen Qin & Zhi-Quan Wu

  2. Institute of Nutritional Sciences, Chinese Academy of Science, Shanghai, 200032, People’s Republic of China

    Zhi-Hui Min

  3. Cancer Center, Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, People’s Republic of China

    Jia Fan

  4. Department of Molecular Pathology, Anderson Cancer Center, University of Texas, Houston, TX, 77030, USA

    Yong Liao

  5. Ruijin Hospital, Shanghai 2nd Medical School, Shanghai Jiaotong University, Shanghai, 200030, People’s Republic of China

    Wu Zhang & Jiang Zhu

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  1. Guo-Ming Shi
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  2. Yang Xu
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  17. Jiang Zhu
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Corresponding author

Correspondence to Jia Fan.

Additional information

Guo-Ming Shi and Yang Xu contributed equally to this work.

This is an original work by all the authors and no previous presentations, reports, or publications contain any material that appears in the article.

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Shi, GM., Xu, Y., Fan, J. et al. Identification of side population cells in human hepatocellular carcinoma cell lines with stepwise metastatic potentials. J Cancer Res Clin Oncol 134, 1155–1163 (2008). https://doi.org/10.1007/s00432-008-0407-1

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  • DOI: https://doi.org/10.1007/s00432-008-0407-1

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