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The effects of growth factors on hIK1 protein expression levels in transfected HEK293 cells
Author(s) -
O’Rourke Kellie Leatha,
Mague Alissa Renee,
Jones Heather Marie
Publication year - 2008
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.22.1_supplement.1201.12
Subject(s) - hek 293 cells , epidermal growth factor , transfection , cell growth , western blot , growth factor , microbiology and biotechnology , biology , cell , cell culture , chemistry , receptor , gene , biochemistry , genetics
The calcium (Ca 2+ )‐activated potassium (K + ) channel, hIK1, is a human intermediate conductance channel expressed in numerous epithelia linings including the placenta, lungs and gastrointestinal tract. The functional roles for hIK1 underline processes such as ion transport, volume regulation, cell growth and proliferation. To investigate hIK1's role in cell growth, we examined the effects of the growth factors: epidermal growth factor (EGF), fibroblast growth factor (FGF), and tumor necrosis growth factor alpha (TNF‐α) on hIK1 protein expression. The protein levels were measured by treating human embryonic kidney (HEK) cells transfected with hIK1. The growth factors were administered 24 hours prior to analysis. Cells were lysed, total protein was collected and equal quantities were run on SDS‐page gel. Antibodies were used to detect hIK1 expression. From the total isolated protein the relative thickness of the treated cell bands on the western blot were compared to the untreated hIK1‐HEK cell bands to determine if total hIK1 protein expression was increased or decreased. It was found that EGF‐treatment increased total hIK1 protein expression and TNF‐α treatment decreased hIK1 protein levels. These results suggest that growth factor interactions with hIK1 are mediating the channel's role in cell growth and proliferation.