Supplementary Figure S2 from Simvastatin-Induced Apoptosis in Osteosarcoma Cells: A Key Role of RhoA-AMPK/p38 MAPK Signaling in Antitumor Activity

Kamel, Walied A.; Sugihara, Eiji; Nobusue, Hiroyuki; Yamaguchi-Iwai, Sayaka; Onishi, Nobuyuki; Maki, Kenta; Fukuchi, Yumi; Matsuo, Koichi; Muto, Akihiro; Saya, Hideyuki; Shimizu, Takatsune

doi:10.1158/1535-7163.22508220.v1

Supplementary Figure S2 from Simvastatin-Induced Apoptosis in Osteosarcoma Cells: A Key Role of RhoA-AMPK/p38 MAPK Signaling in Antitumor Activity

https://doi.org/10.1158/1535-7163.22508220

journal contribution

posted on 2023-04-03, 15:45 authored by Walied A. Kamel, Eiji Sugihara, Hiroyuki Nobusue, Sayaka Yamaguchi-Iwai, Nobuyuki Onishi, Kenta Maki, Yumi Fukuchi, Koichi Matsuo, Akihiro Muto, Hideyuki Saya, Takatsune Shimizu

<p>(A) Immunoblot analysis of AMPK and AKT in AXT cells treated with simvastatin for 12 h. The phosphorylation level of AMPK at Ser485 was not affected by simvastatin, while treatment of 1 μg/ml of insulin for 1 h (Ins) slightly enhanced phosphorylation level of AMPK at Ser485/Ser491. (B) Phase-contrast microscopy of AXT cells exposed to the indicated reagents for 20 h. (C, D) Immunoblot analysis of caspase-3 cleavage and either AMPK (B) or p38 MAPK (C) phosphorylation in AXT cells exposed to the indicated concentrations of 5-iodotubercidin or BIRB796, respectively, in the additional presence of 5 μM simvastatin for 20 h. (E) Immunoblot analysis of AMPK, p38 MAPK, and ACC phosphorylation in AXT cells treated with AICAR for 22 h. (F) Viability of AXT cells exposed to compound C in the additional presence of AICAR for 2 days. (G) Phase-contrast microscopy of AXT cells exposed to C3 transferase for 12 h.</p>

Funding

Japan Agency for Medical Research and Development

MEXT

Takeda Science Foundation

History

Related Materials

1.
DOI - Is supplement to Simvastatin-Induced Apoptosis in Osteosarcoma Cells: A Key Role of RhoA-AMPK/p38 MAPK Signaling in Antitumor Activity

ARTICLE ABSTRACT

Osteosarcoma is the most common type of primary bone tumor, novel therapeutic agents for which are urgently needed. To identify such agents, we screened a panel of approved drugs with a mouse model of osteosarcoma. The screen identified simvastatin, which inhibited the proliferation and migration of osteosarcoma cells in vitro. Simvastatin also induced apoptosis in osteosarcoma cells in a manner dependent on inhibition of the mevalonate biosynthetic pathway. It also disrupted the function of the small GTPase RhoA and induced activation of AMP-activated protein kinase (AMPK) and p38 MAPK, with AMPK functioning upstream of p38 MAPK. Inhibitors of AMPK or p38 MAPK attenuated the induction of apoptosis by simvastatin, whereas metformin enhanced this effect of simvastatin by further activation of AMPK. Although treatment with simvastatin alone did not inhibit osteosarcoma tumor growth in vivo, its combination with a fat-free diet induced a significant antitumor effect that was enhanced further by metformin administration. Our findings suggest that simvastatin induces apoptosis in osteosarcoma cells via activation of AMPK and p38 MAPK, and that, in combination with other approaches, it holds therapeutic potential for osteosarcoma. Mol Cancer Ther; 16(1); 182–92. ©2016 AACR.