Abstract
Purpose: Approximately 20% of breast cancers (BC) among US women overexpress HER2, an oncoprotein that stimulates increased cellular proliferation and survival. HER2 acts through two major signaling pathways, PI3K/Akt and Raf/MAPK, and its overexpression is associated with aggressive disease, resistance to therapy, and poor prognosis. Our laboratory has previously demonstrated in preclinical models that calorie restriction (CR) modulates HER2 BC pathogenesis, decreasing incidence and increasing latency in transgenic MMTV-HER2/neu mice. Our current study aims to determine the impact of CR on HER2 signaling and cancer cell phenotypes, including proliferation, cell cycle progression, and apoptosis. Methods: In vitro experiments were performed using a BC cell line derived from the MMTV-HER2/neu mouse model. To mimic CR, cells were treated with nutrient restricted media containing either reduced glucose (1mM), reduced serum (1%) or both (1mM/1%). The impact of CR on proliferation and survival was measured using growth curve experiments, and flow cytometric analysis of cell cycle and apoptosis. HER2 signaling proteins were assessed by western blotting. Results: Relative to cells grown in control media (25mM glucose/10% BCS): a) cells grown in glucose restricted (1mM) + serum restricted (1%) media grew 87% slower (p<0.01); b) cells grown in serum-restricted media (1%) with control glucose levels (25mm) grew 86% slower (p<0.01); and c) cells grown in glucose restricted media (1mM) with control serum levels (10%) grew 67% slower (p<0.05). Serum restricted groups, regardless of glucose levels, also exhibited increased G1 cell cycle arrest following 24 hours of treatment and increased percentage of apoptotic cells following 72 hours of treatment compared to control. In contrast, glucose restriction alone did not significantly affect apoptosis compared to control; however, it induced a G2/M arrest, possibly explaining reduction in growth. Western blotting analysis revealed that serum restricted cells had reduced levels of pAkt (Ser473) but no differences in Akt, pERK, or ERK, compared to the control and 1mM groups following 24 hours of CR treatment. Conclusions: In vitro models of CR, specifically reduced serum proteins alone or in combination with glucose, reduced activation of PI3K/Akt in HER2 BC cells. This altered signaling was associated with reduced tumorigenic potential, as evidenced by decreased growth and increased apoptosis. These findings suggest that combining a nutrient restriction regimen or pharmacologic mimetic of CR with HER2-targeted therapy, which also inhibits PI3K/Akt signaling, may act synergistically. Future experiments will test the hypothesis that CR modulates the efficacy of HER2 targeted therapies through in vitro and in vivo models of HER2 BC.
Citation Format: Laura A. Smith, Magdalena A. Rainey, Nishita T. Sheth, Ciara H. O'Flanagan, Laura W. Bowers, Stephen D. Hursting. Calorie restriction reduces PI3K/Akt signaling and tumorigenic potential in HER2-overexpressing breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 379.
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