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Overexpression of Granulocyte Macrophage Colony Stimulating Factor in Breast Cancer Cells Leads Towards Drug Sensitization

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Abstract

This report describes the effect of overexpressing granulocyte macrophage colony stimulating factor (GMCSF) in breast cancer cells, which otherwise is involved in proliferation and differentiation of granulocyte and macrophage lineages. The purified recombinant GMCSF cytokine is known to exert dose-dependent proliferative response on various cancer cells, but its effect during overexpression is yet to be evaluated. In our present study, we have generated MCF-7 (breast cancer) cells overexpressing GMCSF. Interestingly, cell viability studies showed pronounced sensitivity of GMCSF overexpressing MCF-7 cells towards anticancer drugs, such as, doxorubicin, 5FU and cisplatin. These findings were substantiated by cell cycle analysis of the drug-treated GMCSF overexpressing MCF-7 cells. Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) results revealed differential expressions of cyclins, and the carboxyfluorescein succinimidyl ester (CFSE)-based assay established decrease in doubling time of GMCSF overexpressed cells with respect to the control populations. Thus, overexpressing of proliferative GMCSF cytokine in breast cancer cells may increase susceptibility to anticancer drugs.

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Acknowledgments

This research work was supported by Department of Biotechnology (no. BT/01/NE/PS/08) and Department of Electronics and Information Technology, Government of India for financial support (no. 5(9)/2012-NANO (Vol. II)). The authors acknowledge the assistance from the Central Instruments Facility (CIF) and the Centre for Nanotechnology, IIT Guwahati.

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Authors and Affiliations

  1. Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India

    Nidhi Chaubey & Siddhartha Sankar Ghosh

  2. Centre for Nanotechnology, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India

    Siddhartha Sankar Ghosh

Authors
  1. Nidhi Chaubey
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  2. Siddhartha Sankar Ghosh
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Correspondence to Siddhartha Sankar Ghosh.

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Research Highlights

1. Granulocyte macrophage colony stimulating factor (GMCSF) expressing breast cancer (MCF-7) cell line was generated.

2. Expression of GMCSF cytokine was confirmed by semiquantitative PCR and Western blot analysis.

3. GMCSF overexpressing cells were found to be sensitive to chemotherapeutic drugs such as 5FU, cisplatin and doxorubicin.

4. Flow cytometry experiments revealed cell cycle arrests during drug treatment on GMCSF overexpressing cancer cells.

5. Cyclin expression profile and reduction in doubling time corroborated cell cycle arrest leading to drug sensitization of GMCSF overexpressing cells.

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Chaubey, N., Ghosh, S.S. Overexpression of Granulocyte Macrophage Colony Stimulating Factor in Breast Cancer Cells Leads Towards Drug Sensitization. Appl Biochem Biotechnol 175, 1948–1959 (2015). https://doi.org/10.1007/s12010-014-1373-5

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  • DOI: https://doi.org/10.1007/s12010-014-1373-5

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