PF-06409577 Activates AMPK Signaling and Inhibits Osteosarcoma Cell Growth.

Yun-Rong Zhu,Xiang-Yang Zhang,Cheng-Jian Yu,Qiu-Ping Wu,Yuan-Yuan Liu,Yun-Qing Zhang

doi:10.3389/fonc.2021.659181

Abstract

Osteosarcoma (OS) is a common primary bone malignancy. We here investigated the potential activity of PF-06409577, a novel, potent, and direct activator of AMP-activated protein kinase (AMPK), against human OS cells. In established (U2OS, MG-63, and SaOs-2 lines) and primary human OS cells, PF-06409577 inhibited cell viability and proliferation, while inducing cell apoptosis and cell cycle arrest. PF-06409577 induced AMPK activation, mTORC1 inhibition, autophagy induction, and downregulation of multiple receptor tyrosine kinase inOS cells. AMPK inactivation by AMPKα1 shRNA, CRISPR/Cas9 knockout, or dominant negative mutation (T172A) was able to abolish PF-06409577-induced activity in OS cells. In vivo, PF-06409577 oral administration at well-tolerated doses potently inhibited growth of U2OS cells and primary human OS cells in severe combined immunodeficient mice. AMPK activation, mTORC1 inhibition, autophagy induction, as well as RTK degradation and apoptosis activation were detected in PF-06409577-treated xenografts. In conclusion, activation of AMPK by PF-06409577 inhibits OS cell growth.

Highlights

Osteosarcoma (OS) is a common primary and malignant bone tumor detected mostly in children, adolescents, and young adults [1]
We have previously shown that AMPK activation by microRNA-135b inhibited human OS cell growth and proliferation [6]
By performing MTT assay, we show that PF06409577 efficiently inhibited U2OS cell viability (Figure 1A)

Summary

INTRODUCTION

Osteosarcoma (OS) is a common primary and malignant bone tumor detected mostly in children, adolescents, and young adults [1]. AMPK negatively regulates aerobic glycolysis and cellular biosynthesis to inhibit human cancer cell growth and proliferation [13]. Overexpression of AMPKa1 can suppress human cancer cell growth and proliferation [17]. Existing studies have confirmed that forced activation of AMPK cascade can efficiently inhibit human OS cell growth [22,23,24,25]. Fan et al revealed that 6-gingerol, by activating AMPK cascade, induced proliferation inhibition and apoptosis in OS cells [24]. We have previously shown that AMPK activation by microRNA-135b inhibited human OS cell growth and proliferation [6]. These results indicated that activating AMPK signaling can induce profound OS cell inhibitory activity. We show that the activation of AMPK by PF06409577 inhibited human OS cell growth in vitro and in vivo

MATERIAL AND METHODS

RESULTS

DISCUSSION

ETHICS STATEMENT