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Evaluating the Role of NHE1 Palmitoylation in the Regulation of Cell Proliferation and Migration
Author(s) -
Hanowski Stephanie A.,
Provost Joseph J.,
Wallert Mark A.
Publication year - 2019
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.2019.33.1_supplement.476.10
Subject(s) - palmitoylation , hek 293 cells , cell growth , microbiology and biotechnology , cell migration , embryonic stem cell , cancer cell , chemistry , cancer research , biology , cell , cell culture , cancer , biochemistry , cysteine , gene , genetics , enzyme
The Na + ‐ H + Exchanger Isoform I (NHE1) is a key regulator of cell proliferation and migration in a range of cell types. Activation of NHE1 is an initial step in the development of the transformed phenotypes of cancer cells and therefore a key driver for tumor development and cancer progression. NHE1 also contributes to two of the hallmarks of cancer, the activation of invasion and metastasis and sustained proliferative growth. We have identified for the first time, that NHE1 undergoes reversible palmitoylation, the addition of a palmitic acid residue to the cytoplasmic regulatory domain of NHE1. This suggests that targeting the potential palmitoylation sites on NHE1 may provide a therapeutic target for cancer treatment. This research evaluates the role of NHE1 palmitoylation in two cell lines: 1) Chinese hamster lung fibroblasts expressing human NHE1 (PSN) and 2) Human embryonic kidney (HEK293). The HEK293 were initiated by the transformation and culturing of normal HEK cells with sheared adenovirus 5 DNA in order to employ CRISPR‐CAS 9 gene‐editing techniques. The HEK293 cells were analyzed in the same ways the PSNs were to show a correlation within the different conditions being experimented with. We hypothesize the process of inhibiting palmitoylation will alter regulation of NHE1 and will play a role in the control of cell proliferation and migration. Using the palmitoylation inhibitor, 2‐bromopalmitate (2BP), we evaluated cell growth and migration in the presence and absence of palmitoylation. We also evaluated cell growth and migration in the presence and absence of the chemical inhibitor of NHE1, Cariporide (C). In proliferation assays, the rate of cell proliferation increased in both cell lines by approximately 5‐fold when the serum present in the growth media was increased from 0.5% to 10%. Under both conditions, the addition of 15 μM 2BP or 10 μM cariporide dramatically reduced or eliminated the increased proliferation. We also evaluated the rate of cell migration and once again, the rate was reduced by 50% or more in the presence of 15 μM 2BP or 10 μM cariporide. These data demonstrated that inhibiting the palmitoylation of NHE1 decreases both cell proliferation and the rate of cell migration. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .