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HepG2 cells acquire stem cell-like characteristics after immune cell stimulation

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Abstract

Background

The presence of cancer stem cells (CSCs) is currently regarded as one of the main culprits of tumor formation and therapy failure. It is known that chronic inflammation is associated with CSCs, but it is not clear yet how inflammation affects the development of CSCs. In the present study we aimed to examine the relationship between cancer cell stimulation mediated by immune cells and the acquisition of a CSC-like phenotype.

Methods

Cancer cells derived from single hepatocarcinoma HepG2 cells were treated with mouse splenic B cells (MSBCs) and mouse peritoneal macrophage cells (MPMCs), respectively. The stem cell-like characteristics of the resulting HepG2 cells (MSBC-HepG2 and MPMC-HepG2) were evaluated using different assays, including biomarker assays, in vitro tumoroid and colony forming assays, in vivo tumor forming assays and signal transduction pathway activation assays.

Results

Various stemness characteristics of HepG2 cells, including self-renewal, proliferation, chemoresistance and tumorigenicity were evaluated. The expression levels of stemness-related genes and its encoded proteins in the MSBC-HepG2 and MPMC-HepG2 cells were assessed using RT-PCR and FACS analyses. We found that MSBC-HepG2 and MPMC-HepG2 cells possess hepatic CSC properties, including persistent self-renewal, extensive proliferation, drug resistance, high tumorigenic capacity and over-expression of CSC-related genes and proteins (i.e., EpCAM, ALDH, CD133 and CD44), compared to the parental cells. We also found that 1x103 MSBC-HepG2 and MPMC-HepG2 cells were able to form tumors in NOD/SCID mice and that the Notch and SHH signaling pathways were highly activated in MSBC-HepG2 cells.

Conclusions

We conclude that the immune system may have a double-edge effect on cancer development. On one hand, immune cells such as B lymphocytes and macrophages may recognize, attack and eliminate cancer cells, whereas on the other hand, they may promote a subset of cancer cells to acquire stem cell-like characteristics.

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Acknowledgments

This study was supported by the Fujian Minfaigaoji foundation (NO. 2014–514), the Fujian Education Office (NO. JA12033) and the Fuzhou University (NO.2013-XQ-27), Fujian province, P.R. China.

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Authors and Affiliations

  1. Institute of Pharmaceutical Biotechnology and Engineering, College of Biological Science and Biotechnology, Fuzhou University, Fuzhou, Fujian, 350108, China

    Hang Wang, Miqing Yang, Ling Lin, Hongzhen Ren, Chaotong Lin, Suling Lin, Guoying Shen, Binfeng Ji & Chun Meng

  2. Qi Shan Campus, 2 Xue Yuan Road, University Town, Fuzhou, Fujian, 350108, People’s Republic of China

    Chun Meng

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  1. Hang Wang
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  2. Miqing Yang
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  3. Ling Lin
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  4. Hongzhen Ren
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Correspondence to Chun Meng.

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Hang Wang and Miqing Yang are Joint first authors

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Wang, H., Yang, M., Lin, L. et al. HepG2 cells acquire stem cell-like characteristics after immune cell stimulation. Cell Oncol. 39, 35–45 (2016). https://doi.org/10.1007/s13402-015-0249-1

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