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Analysis of cell cycle, cell death and EGFR expression in hepatic cells WRL68 treated with ethanol
Author(s) -
AlvarezAyala Elizabeth,
ReyesEsparza Jorge,
RodriguezFragoso Lourdes
Publication year - 2010
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.24.1_supplement.565.8
Subject(s) - viability assay , cell cycle , apoptosis , flow cytometry , chemistry , cell growth , cell , cell culture , mtt assay , ethanol , necrosis , western blot , glutamine , hepatic stellate cell , microbiology and biotechnology , cell cycle checkpoint , biology , biochemistry , endocrinology , amino acid , genetics , gene
Our aim was to study the effect of the ethanol on viability, proliferation and EFGR expression in the hepatic cells (WRL68). For the study, we used the WRL‐68 cells (ATCC: CL‐48) cultured in MEM supplemented with 10 % fetal bovine serum, 2mol/L glutamine, 1% non essential amino acid. Cells were kept at 37°C, under an atmosphere 95% of air and 5% CO2. We treated hepatic cells with ethanol at concentrations of 5, 12, 25, 50, 100, 200, 400 and 800 mg/dL. The viability and cell cycle was analyzed using a MTT assay. Cell cycle and apoptosis were analyzed at different times (cell cycle at 9, 12, 15 and 24 h, and apoptosis at 24 h) by flow cytometry. EGFR expression was analyzed by western‐blot. The ethanol did not modify the viability and cell proliferation. Ethanol induced changes on S phase at 9–15h but did not alter the Go/G1 and G2+M phases in WRL68 cells. Ethanol induces death cell, we observed the presence of apoptotic (40%) and necrotic cells (10%). We observed the EGFR expression in the hepatic cell line WRL68. However, no changes in EGFR expression were observed in cells treated with ethanol. In conclusion, ethanol induces apoptosis, necrosis and changes in cell cycle, but not changes in EGFR expression were observed in hepatic cells WRL‐68.