Abstract 153: Honokiol, a promising small molecular weight natural agent, activates AMP-activated protein kinase in breast cancer cells via LKB1-dependent pathway and inhibits breast carcinogenesis
Authors: Nagalingam Arumugam , Arbiser Jack L , Bonner Michael Y. , Saxena Neeraj , Sharma Dipali 
Published: August 2012

Abstract
Introduction: Research over the past few years have established LKB1-AMPK pathway as an important metabolic checkpoint interconnecting the signaling pathways controlling metabolism and cancer cell growth. Thus, it is of major interest to find effective therapeutic strategies to manipulate LKB1-AMPK axis to inhibit cancer growth. The importance of active constitutive agents in natural products has become increasingly apparent owing to their potential cancer preventive as well as therapeutic properties. Honokiol, a small-molecule polyphenol isolated from magnolia species, is widely known for its therapeutic potential as an anti-inflammatory, anti-thrombosis, anti-oxidant agent and more recently for its protective function in carcinogenesis. In the present study, we sought to examine the effectiveness of honokiol in modulating LKB1-AMPK axis, preventing mammary tumorigenesis and inhibiting metastasis. Methods: Clonogenicity, 3D-colony formation, matrigel invasion, scratch-migration and spheroid-migration assays were used to examine breast cancer cell growth and metastatic potential upon honokiol treatment. Western blot and immunofluorescence analysis were used to examine activation of LKB1-AMPK axis. Functional importance of AMPK and LKB1 was examined by using AMPK-null and AMPK-WT immortalized mouse embryonic fibroblasts (MEFs) and isogenic LKB1-knockdown cell line pairs. Breast cancer xenografts, immunohistochemical and western blot analysis of tumors were used. Results: Analysis of the underlying molecular mechanisms revealed that honokiol treatment increases AMP-activated protein kinase (AMPK) phosphorylation and activity as evident by increased phosphorylation of downstream target of AMPK, acetyl-coenzyme A carboxylase (ACC) and inhibition of phosphorylation of p70S6kinase (pS6K) and eukaryotic translation initiation factor 4E binding protein 1 (4EBP1). Using AMPK-null and AMPK-WT (MEFs); we found that AMPK is required for honokiol-mediated modulation of pACC-pS6K. Intriguingly, we discovered that honokiol treatment increased the expression and cytoplasmic translocation of tumor suppressor gene LKB1 in breast cancer cells. LKB1 knockdown inhibited honokiol-mediated activation of AMPK and more importantly, inhibition of migration and invasion of breast cancer cells. Furthermore, honokiol treatment resulted in inhibition of breast tumorigenesis in vivo. Analysis of tumors showed significant increase in the levels of cytoplasmic LKB1 and phospho-AMPK in honokiol-treated tumors. Conclusions: Taken together, these data providing first in vitro and in vivo evidence of the efficacy of honokiol in activating LKB1-AMPK axis resulting in effective inhibition of mammary tumorigenesis and metastasis provide impetus to determine its activity in clinical setting.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 153. doi:1538-7445.AM2012-153