Molecular Mechanisms of Cigarette Smoke-Induced Proliferation of Lung Cells and Prevention by Vitamin C

Figure 10

Exposure of A549 cells to AECS/p-BQ results in overexpression of Hras + Kras and activation of downstream proliferation and survival signaling Akt and ERK 1/2. Serum-starved A549 cells were either nontreated (NT) or exposed to 2 μL/mL AECS or 200 ng/mL p-BQ for 1 hr followed by incubation in serum containing media for 6 hr. After 6 hr, cells were lysed, and cell lysates were separated by SDS-PAGE, transferred to PVDF membrane, and immunoblotted with antiphosphotyrosine (PY20), anti-HRAS + KRAS, anti-phospho-Akt, anti-Akt, anti-phospho-ERK 1/2, and anti-ERK 1/2 antibodies. β-actin was used as the loading control.