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NK-1 receptor antagonists induce apoptosis and counteract substance P-related mitogenesis in human laryngeal cancer cell line HEp-2

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Summary

It has been demonstrated that substance P (SP) induces cell proliferation and neurokinin-1 (NK-1) receptor antagonists inhibit growth in several human cancer cell lines, but it is currently unknown whether such actions are exerted on human laryngeal carcinoma cell line HEp-2. In addition, the presence of NK-1 receptor has not been demonstrated in this cell line. We carried out an in vitro study of the growth inhibitory capacity of the NK-1 receptor antagonists L-733,060 and L-732,138 against human laryngeal carcinoma cell line HEp-2. Coulter counter was used to determine viable cell numbers followed by application of the tetrazolium compound MTS. Furthermore, an immunoblot analysis was used to determine the NK-1 receptor, and the 4′,6-diamidino-2-phenylindole (DAPI) method was applied to demonstrate apoptosis of the laryngeal carcinoma cells. We observed the presence of several NK-1 receptors isoforms (34, 46, 58 and 75 kDa). Nanomolar concentrations of SP increased the growth rate of the cell line and micromolar concentrations of L-733,060 and L-732,138 inhibited the growth of the HEp-2 cells in a dose-dependent manner, with and without previous administration of SP. The 50% inhibition concentration values were 21.34 μM and 37.97 (48 h) respectively for HEp-2. NK-1 receptor presence on HEp-2 cells was confirmed by western blotting. DAPI staining revealed the presence of apoptosis following NK-1 receptor antagonists treatment. We demonstrated that NK-1 receptors were present in this laryngeal cancer cell line; these findings demonstrate that SP acts as a mitogen on the human laryngeal carcinoma cell line HEp-2 through the NK-1 receptor, and also indicate that both NK-1 receptors antagonists induced apoptosis of the tumour cells. This new action, reported here for the first time, suggests that the NK-1 receptor is a new and promising target in the treatment of human laryngeal carcinoma.

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Acknowledgements

The authors thank Manuel Sánchez for technical assistance.

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Authors and Affiliations

  1. Hospital Universitario Virgen del Rocío, Seville, Spain

    Miguel Muñoz, Marisa Rosso, Francisco J Aguilar & Francisco Esteban

  2. University of Granada, Granada, Spain

    Miguel A González-Moles

  3. Departament of Biochemistry, Hospital Costa del Sol, Marbella, Spain

    Maximino Redondo

  4. Unidad de Cuidados Intensivos Pediátricos, Hospital Infantil Virgen del Rocío, Av. Manuel Siurot s/n, 41013, Seville, Spain

    Miguel Muñoz

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  1. Miguel Muñoz
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Correspondence to Miguel Muñoz.

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Muñoz, M., Rosso, M., Aguilar, F.J. et al. NK-1 receptor antagonists induce apoptosis and counteract substance P-related mitogenesis in human laryngeal cancer cell line HEp-2. Invest New Drugs 26, 111–118 (2008). https://doi.org/10.1007/s10637-007-9087-y

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