Abstract 2765: Metformin represses cancer cells via alternate pathways in N-Cadherin wild-type and N-Cadherin-deficient cells

Abstract

Abstract BACKGROUND Metformin, one of the most commonly used medications for treatment of type 2 diabetes, has emerged as a potential anticancer agent. The molecular mechanisms associated with the antitumor effect of metformin are still poorly understood. In this study, we show that Metformin represses cancer cells via alternate pathways in N-Cadherin Wild type and N-Cadherin deficient cells. METHODS Cell viability and apoptosis were determined by MTT, TUNEL and FACS. Various stable cell lines were generated in vitro and then injected into both flanks of nude mice to generate xenografts. Mice were treated with i.p. doxorubicin (every 5 days × 4 cycles with 4 mg/kg) or p.o.metformin (200 ug/ml, diluted in the drinking water). Xenograft tumor volume was measured every 5 days. RESULTS We found that metformin represses N-cadherin, independent of AMPK, in wild-type N-cadherin cancer cells. Ectopic expression of N-cadherin rendered cells resistant to metformin. Silencing or over-expression of AMPK did not alter sensitivity of cancer cells to metformin in N-cadherin wild-type cells. In metformin-resistant cancer cells, N-cadherin level did not change with metformin treatment. Suppression of N-cadherin changed the phenotype of metformin-resistant to metformin-sensitive cells. We also demonstrated that NF-kB is a downstream molecule of N-cadherin and metformin regulated NF-kB signaling via inhibiting N-cadherin, rather than direct modulation of NF-kB. Silencing of NF-kB in resistant cells sensitized the cells to metformin. We also found that metformin down-regulated N-cadherin and TWIST1 concurrently. Ectopic expression of TWIST1 regulated N-cadherin/NF-kB signaling and made cells more resistant to metformin. Suppression of TWIST1 sensitized resistant cells to metformin. Metformin inhibited growth of tumor xenografts, and high levels of N-cadherin made tumors more resistant to metformin, while suppression of N-cadherin made tumors more sensitive. In N-cadherin deficient cancer cells, metformin activated AMPK. Suppression of AMPK compromised metformin-mediated inhibition of NF-kB and changed the cells to a more metformin-resistant phenotype. In the deficient cells, Ectopic expression of N-cadherin attenuated AMPK's ability to suppress NF-kB, leading to a more metformin resistant phenotype. Similarly, a specific AMPK inhibitor, compound C, inhibited the therapeutic effects of metformin. Similar to the in vitro findings, suppression of AMPK in xenografts rendered tumors resistant to metformin. CONCLUSIONS: We suggest that metformin's anti-cancer therapeutic effect in N-cadherin wild-type cells may be mediated through repression of the TWIST/N-cadherin/NF-kB signaling pathway, independent of AMPK. However, in N-cadherin deficient cells, metformin activates AMPK. Elucidating the molecular pathways responsible for metformin's anti-cancer effect may help delineate its role for cancer therapies. Citation Format: Ge Rongbin, Zongwei Wang, Jijun Li, Aria Olumi. Metformin represses cancer cells via alternate pathways in N-Cadherin wild-type and N-Cadherin-deficient cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2765. doi:10.1158/1538-7445.AM2014-2765