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Inhibition of mouse embryonic carcinoma cell growth by lidamycin through down-regulation of embryonic stem cell-like genes Oct4, Sox2 and Myc

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Summary

Lidamycin (LDM, also known as C-1027) as an anti-cancer agent inhibits growth in a variety of cancer cells by inducing apoptosis and cell cycle arrest. In this study we demonstrated that inhibition of mouse embryonic carcinoma (EC) cell growth using LDM at low concentrations can be attributed to a loss of the cell’s self-renewal capability but not to apoptosis or cell death, which can be correlated to the down-regulation of embryonic stem (ES) cell-like genes Oct4, Sox2 and c-Myc. MTT assays showed that LDM inhibited the growth of mouse P19 EC cells in a time- and dose-dependent manner. The EC cells exposed to a low dose (0.01 nM) of LDM lost their capability to generate colonies, as evidenced by the colony forming assay. Flow cytometer analyses demonstrated that LDM induced G1 arrest in exposed EC cells without apoptosis. Real-time qPCR, Western blotting and immunocytochemistry revealed that Oct4, Sox2 and c-Myc were down-regulated in LDM-exposed EC cells, but not adriamycin (ADM)-exposed cells. Furthermore, a combination of the low dose of LDM and ADM significantly reduced the proliferation of the cancer cells than single-agent treatment. This suggested that synergy of ADM and LDM improved chemotherapy. Taking together, our results indicate that LDM can reduce the capability for self-renewal that mouse EC cells possess through the repression of ES cell-like genes, thereby inhibiting carcinoma cell growth. This data also suggests that LDM might have potential for application in CSC-based therapy and be a useful tool for studying ES cell pluripotency and differentiation.

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Acknowledgements

We would like to thank Yong-Shu Zhen (Professor, Department of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Science, Beijing, China) for kindly providing us with the anti-cancer drug lidamycin, Hong-Ti Jia (Professor, Department of Molecular and Biochemistry, Peking University Health Science Center) for the discussion of the manuscript, Ling-Song Li (Chairman, Department of Cell Biology, Peking University Health Science Center) for some financial support and the experimental facility, Prof. Tai-Ping Wang for helping to proofread the manuscript and Dr. Chris Capel, Urs. Lienert and Qimin Huang for the final revision. We are gratefully indebted to Lan Yuan and Ji-Hong Liu for technical support for the confocal microscope. This work was also supported by the Foundation of “211 Project” and “985 Project” in P R China.

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Authors and Affiliations

  1. Department of Cell Biology, Peking University Health Sciences Center, Xue Yuan Road 38, Beijing, 100191, People’s Republic of China

    Hong-Ying Zhen, Shu-Ling Wang, Yi-Nan Liu, Wei-Hua Wu, Xiao-Yan Zhang, Ai-Li Lu & Li Shen

  2. Medical Analysis Center, Peking University Health Sciences Center, Beijing, People’s Republic of China

    Qi-Hua He

  3. Department of Histology and Embryology, North China Coal Medical University, Tangshan, 063000, People’s Republic of China

    Yong-Zhan Zhen

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  1. Hong-Ying Zhen
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Correspondence to Hong-Ying Zhen or Li Shen.

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Hong-Ying Zhen and Qi-Hua He contributed equally to this study.

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Fig. 1

Low dose of LDM maintenance of P19 EC cell growth in non-apoptotic status. As described in the Methods section, the cells were exposed to 0.01 nM LDM for the indicated number of hours and collected from the medium and dishes by centrifuge, separately. Apoptosis assay was carried out as Fig. 4a described by flow cytometer (GIF 312 kb)

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Zhen, HY., He, QH., Zhen, YZ. et al. Inhibition of mouse embryonic carcinoma cell growth by lidamycin through down-regulation of embryonic stem cell-like genes Oct4, Sox2 and Myc. Invest New Drugs 29, 1188–1197 (2011). https://doi.org/10.1007/s10637-010-9463-x

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