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TM9SF4 is a novel V-ATPase-interacting protein that modulates tumor pH alterations associated with drug resistance and invasiveness of colon cancer cells

Oncogene volume 34, pages 5163–5174 (2015)Cite this article

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Abstract

An inverted pH gradient across the cell membranes is a typical feature of malignant cancer cells that are characterized by extracellular acidosis and cytosol alkalization. These dysregulations are able to create a unique milieu that favors tumor progression, metastasis and chemo/immune-resistance traits of solid tumors. A key event mediating tumor cell pH alterations is an aberrant activation of ion channels and proton pumps such as (H+)-vacuolar-ATPase (V-ATPase). TM9SF4 is a poorly characterized transmembrane protein that we have recently shown to be related to cannibal behavior of metastatic melanoma cells. Here, we demonstrate that TM9SF4 represents a novel V-ATPase-associated protein involved in V-ATPase activation. We have observed in HCT116 and SW480 colon cancer cell lines that TM9SF4 interacts with the ATP6V1H subunit of the V-ATPase V1 sector. Suppression of TM9SF4 with small interfering RNAs strongly reduces assembly of V-ATPase V0/V1 sectors, thus reversing tumor pH gradient with a decrease of cytosolic pH, alkalization of intracellular vesicles and a reduction of extracellular acidity. Such effects are associated with a significant inhibition of the invasive behavior of colon cancer cells and with an increased sensitivity to the cytotoxic effects of 5-fluorouracil. Our study shows for the first time the important role of TM9SF4 in the aberrant constitutive activation of the V-ATPase, and the development of a malignant phenotype, supporting the potential use of TM9SF4 as a target for future anticancer therapies.

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Acknowledgements

We thank Dr Angelo De Milito for helpful discussion, Pasqualina Leone for her contribution in plasmid cloning and Dr Tonino Sofia for his suggestions. This study was supported by AIRC Associazione Italiana per la Ricerca sul Cancro. Airc Grant '10602' and ‘IG 11505n’. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

Author Contributions

FL conceived the study, designed and supervised the experiments; MB, FM, GV, SF and AC provided critical advice and contributed to manuscript revisions; MB, FM, EI, TA, PM, SM and RM performed experiments; AC performed quantitative reverse transcription–PCR experiments; TA performed statistical analyses; and FL wrote the manuscript.

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Authors and Affiliations

  1. Therapeutic Research and Medicines Evaluation Department, Istituto Superiore di Sanità, Rome, Italy

    F Lozupone, F Marzoli, T Azzarito, P Matarrese, E Iessi, G Venturi, A Canitano, R Bona, A Cara & S Fais

  2. Parasitic and Immune-Mediated Diseases Department, Infectious, Istituto Superiore di Sanità, Rome, Italy

    M Borghi

  3. Technology and Health Department, Istituto Superiore di Sanità, Rome, Italy

    S Meschini

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  1. F Lozupone
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Correspondence to F Lozupone.

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Lozupone, F., Borghi, M., Marzoli, F. et al. TM9SF4 is a novel V-ATPase-interacting protein that modulates tumor pH alterations associated with drug resistance and invasiveness of colon cancer cells. Oncogene 34, 5163–5174 (2015). https://doi.org/10.1038/onc.2014.437

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