Hydrophobic statins induce autophagy in cultured human rhabdomyosarcoma cells

doi:10.1016/j.bbrc.2007.12.166

Biochemical and Biophysical Research Communications

Volume 367, Issue 2, 7 March 2008, Pages 462-467

https://doi.org/10.1016/j.bbrc.2007.12.166 Get rights and content

Abstract

Statins are widely used to treat hypercholesterolemia, but they are associated with muscle-related adverse events, by as yet, inadequately resolved mechanisms. In this study, we report that statins induced autophagy in cultured human rhabdomyosarcoma A204 cells. Potency differed widely among the statins: cerivastatin induced autophagy at 0.1 μM, simvastatin at 10 μM but none was induced by pravastatin. Addition of mevalonate, but not cholesterol, blocked induction of autophagy by cerivastatin, suggesting that this induction is dependent on modulation of isoprenoid metabolic pathways. The statin-induced autophagy was not observed in other types of cells, such as human hepatoma HepG2 or embryonic kidney HEK293 cells. Muscle-specific abortive induction of autophagy by hydrophobic statins is a possible mechanism for statin-induced muscle-related side effects.

Section snippets

Materials and methods

Materials. Simvastatin and pravastatin were purchased from Wako (Tokyo, Japan) and cerivastatin from Sequoia Research Products (Pangbourne, UK). Mevalonolactone was purchased from Tokyo-Kasei (Tokyo, Japan). Expression plasmid of GFP-LC3 (microtubule-associated protein 1 light chain 3, an autophagy marker protein) [10] was a gift from Prof. N. Mizushima (Tokyo Medical and Dental University, Tokyo, Japan).

Cell culture and transfection. Human rhabdomyosarcoma A204 cells were obtained from ATCC

Cerivastatin dose-dependently induces cellular autophagy

To examine the possibility that statins induce autophagy in muscle, we first established a human rhabdomyosarcoma cell line (A204) that constitutively expressed LC3 fused with GFP [10]. The fluorescent GFP-LC3 on autophagosome membranes that had been formed during autophagy was investigated with confocal microscopy to monitor the induction of autophagy. The cells inducing autophagy by starvation by incubating in Hank’s balanced salt solution (HBSS) served as a positive control, and the

Discussion

Microscopic observation and biochemical analysis revealed that hydrophobic cerivastatin and simvastatin but not hydrophilic pravastatin induced autophagy in cultured human rhabdomyosarcoma cells in a dose-dependent manner (Fig. 1). When compared with previous studies in which adverse effects of statins were investigated, the statin concentrations used in this study were low and the incubation period to detect the side effects was short: 0.1–1.0 μM cerivastatin in 12 h in this study vs 100 μM for 24

Acknowledgments

This work was supported in part by Meiyaku Open Research Project and Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science.

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Citation Excerpt :
This selective statin-induced autophagy is due to inhibition of cholesterol synthesis and counteracts concomitant cholesterol depletion-dependent apoptotic demise of leukemic cells. Recently, we and others have shown that various statins, including simvastatin, lovastatin, atorvastatin, fluvastatin, pravastatin, cerivastatin, and rosuvastatin induce autophagy in hepatocellular and colorectal carcinoma, glioma, mesothelioma, rhabdomyosarcoma, prostate, thyroid, bladder and ovarian cancer cell lines (Araki et al., 2012; Araki and Motojima, 2008; Asakura et al., 2011; He et al., 2012; Jiang et al., 2014; Kang et al., 2014; Misirkic et al., 2012; Parikh et al., 2010; Robinson et al., 2013; Toepfer et al., 2011; Yang et al., 2010; Zeybek et al., 2011), as well as in normal fibroblasts and smooth muscle cells (Ghavami et al., 2011; Ghavami et al., 2014, 2012; Wei et al., 2013). As statin-induced autophagy has thus far not been studied in leukemic cells, we screened a panel of human cell lines, primary tumor samples, and normal human leukocytes to determine their sensitivity to statin treatment.
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Hydrophobic statins induce autophagy in cultured human rhabdomyosarcoma cells

Abstract

Section snippets

Materials and methods

Cerivastatin dose-dependently induces cellular autophagy

Discussion

Acknowledgments

J. Am. Coll. Cardiol.

Toxicol. Appl. Pharmacol.

Int. J. Pharm.

Biochem. Biophys. Res. Commun.

Trends Biochem. Sci.

Cell Metab.

Cell Metab.

Inhibition of cholesterol synthesis in vitro and in vivo by ML-236A and ML-236B, competitive inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase

Eur. J. Biochem.

Statin-associated myopathy

Jama

FDA adverse event reports on statin-associated rhabdomyolysis

Ann. Pharmacother.

Statin-induced myopathy: the two faces of Janus

J. Cardiovasc. Pharmacol. Ther.

A metabolic switching hypothesis for the first step in the hypolipidemic effects of fibrates

Biol. Pharm. Bull.