Rat mammary epithelial cells line: A biphasic activation of extracellular signal-regulated kinase (ERK) 1/2
محورهای موضوعی : کلینیکال پاتولوژی
1 - Department of Medical Laboratory Sciences, Urmia Branch, Islamic Azad University, Urmia, Iran
کلید واژه: Rat, Biphasic, ERK 1/2, Phosphorylation,
چکیده مقاله :
Animal models help scientists evaluate the molecular basis of cancer and explore new treatment options. ERK 1/2, a cell signaling component, is a critical molecule in pathogenesis and an important target for breast cancer. In the brain and some other tissues, ERK 1/2 phosphorylation shows a biphasic time-dependent activation. We hypothesize that ERK 1/2 phosphorylation exhibits a similar pattern in rat breast cell lines. The rat mammary epithelial cells (RMECs) were isolated and exposed to RPMI 1640 containing 5%, 10%, and 15% rat serum, with serum-free media as a negative control. Using the WST-1 technique, we evaluated cell proliferation. The cells were exposed to 10% serum-enriched media, and the cell lysate was treated with RIPA buffer in a time-dependent manner (1, 5, 10, 15, and 30 minutes). ERK 1/2 phosphorylation was evaluated by Western blot using antibodies against total and phosphorylated rat ERK 1/2 proteins. The cell proliferation assay showed that serum at a concentration over 5% can induce cell proliferation in RMECs. Serum media at concentrations higher than 10% had the same effect on cell proliferation. ERK1/2 proteins were phosphorylated 5 minutes after exposure, underwent dephosphorylation after 10 minutes, and rephosphorylation occurred from 15 to 30 minutes as a second phase. Taken together, we can infer that the behavior of cell signaling components, especially the ERK1/2 phosphorylation pattern, differs from species to species.
Animal models help scientists evaluate the molecular basis of cancer and explore new treatment options. ERK 1/2, a cell signaling component, is a critical molecule in pathogenesis and an important target for breast cancer. In the brain and some other tissues, ERK 1/2 phosphorylation shows a biphasic time-dependent activation. We hypothesize that ERK 1/2 phosphorylation exhibits a similar pattern in rat breast cell lines. The rat mammary epithelial cells (RMECs) were isolated and exposed to RPMI 1640 containing 5%, 10%, and 15% rat serum, with serum-free media as a negative control. Using the WST-1 technique, we evaluated cell proliferation. The cells were exposed to 10% serum-enriched media, and the cell lysate was treated with RIPA buffer in a time-dependent manner (1, 5, 10, 15, and 30 minutes). ERK 1/2 phosphorylation was evaluated by Western blot using antibodies against total and phosphorylated rat ERK 1/2 proteins. The cell proliferation assay showed that serum at a concentration over 5% can induce cell proliferation in RMECs. Serum media at concentrations higher than 10% had the same effect on cell proliferation. ERK1/2 proteins were phosphorylated 5 minutes after exposure, underwent dephosphorylation after 10 minutes, and rephosphorylation occurred from 15 to 30 minutes as a second phase. Taken together, we can infer that the behavior of cell signaling components, especially the ERK1/2 phosphorylation pattern, differs from species to species.
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