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Characterizing the role of the Na + ‐ H + Exchanger Isoform 1 (NHE1) in cell proliferation and migration in ovarian cancer cells.
Author(s) -
Manzella Taylor M,
Provost Joseph J,
Wallert Mark A.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.934.17
Subject(s) - cell growth , protein kinase b , chemistry , microbiology and biotechnology , cell culture , lysophosphatidic acid , pi3k/akt/mtor pathway , cancer research , signal transduction , biology , biochemistry , receptor , genetics
To sustain proliferative growth, cancer cells exhibit disrupted signaling pathways that compromise the ability of the cell to regulate homeostasis. The Na + ‐ H + Exchanger Isoform 1 (NHE1) is a transmembrane protein activated in cancer cells that plays a role in sustaining proliferative growth. In this study, three ovarian adenocarcinoma lines, SKOV‐3, OVCAR‐3, and CAOV‐3 have been characterized to determine the role that NHE1 plays in their growth and progression. We have used proliferation assays to demonstrate NHE1 involvement in the regulation of cell growth in ovarian cancer cell lines. Cells growth is stimulated when cells are cultured in 10% serum compared to 0.5% serum and this growth stimulation is blocked by the NHE1 inhibitor Cariporide. Additionally, we investigated the effects of four inhibitors that block specific kinases that phosphorylate NHE1 to evaluate their impact on cell proliferation. Specific inhibitors, MK2206 (AKT), Sch772984 (ERK), Y27632 (Rock), and B‐ID1870 (RSK) were evaluated in the three ovarian cell lines to determine key pathways in regulating NHE1. Amongst the SKOV‐3 and CAOV‐3 cell lines, the RSK site inhibitor, B‐ID1870 demonstrated the largest impact of inhibition while in the OVCAR‐3 cell line, the AKT site inhibitor, MK2206, demonstrated the greatest impact. In addition to serum stimulation we are evaluating the ability of lysophosphatidic acid (LPA) and urokinase‐type plasminogen activator (uPA) to activate NHE1 and stimulate proliferation in these cells. Further investigation of the role of NHE1 will include preparing an NHE1 knock out cell line using CRISPR‐CAS9 gene editing technology as well as the evaluation of the role of NHE1 in invasion and metastasis using ECIS migration assays to define cell growth.