Simvastatin Induces Unfolded Protein Response and Enhances Temozolomide-Induced Cell Death in Glioblastoma Cells
Authors: Afshin Samali
Pawan Sharma
Pooneh Mokarram
Saeid Ghavami
Shahla Shojaei
Sanaz Dastghaib
John B Patterson
Zohreh Mostafavi-Pour
Journal: Cells (2020)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: ER stress; autophagy; autophagy flux; glioblastoma; mevalonate cascade
Affiliations: Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz 7134845794, Iran., Department of Human Anatomy and Cell Science, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, Winnipeg, MB R3E 0J9, Canada., Center for Translational Medicine, Division of Pulmonary, Allergy and Critical Care Medicine, Jane & Leonard Korman Respiratory Institute, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA., Orinove, Newbury Park, CA 91320, USA., Apoptosis Research Centre, National University of Ireland, H91 W2TY Galway, Ireland.

Abstract:  Glioblastoma (GBM) is the most prevalent malignant primary brain tumor with a very poor survival rate. Temozolomide (TMZ) is the common chemotherapeutic agent used for GBM treatment. We recently demonstrated that simvastatin (Simva) increases TMZ-induced apoptosis via the inhibition of autophagic flux in GBM cells. Considering the role of the unfolded protein response (UPR) pathway in the regulation of autophagy, we investigated the involvement of UPR in Simva-TMZ-induced cell death by utilizing highly selective IRE1 RNase activity inhibitor MKC8866, PERK inhibitor GSK-2606414 (PERKi), and eIF2α inhibitor salubrinal. Simva-TMZ treatment decreased the viability of GBM cells and significantly increased apoptotic cell death when compared to TMZ or Simva alone. Simva-TMZ induced both UPR, as determined by an increase in GRP78, XBP splicing, eukaryote initiation factor 2α (eIF2α) phosphorylation, and inhibited autophagic flux (accumulation of LC3β-II and inhibition of p62 degradation). IRE1 RNase inhibition did not affect Simva-TMZ-induced cell death, but it significantly induced p62 degradation and increased the microtubule-associated proteins light chain 3 (LC3)β-II/LC3β-I ratio in U87 cells, while salubrinal did not affect the Simva-TMZ induced cytotoxicity of GBM cells. In contrast, protein kinase RNA-like endoplasmic reticulum kinase (PERK) inhibition significantly increased Simva-TMZ-induced cell death in U87 cells. Interestingly, whereas PERK inhibition induced p62 accumulation in both GBM cell lines, it differentially affected the LC3β-II/LC3β-I ratio in U87 (decrease) and U251 (increase) cells. Simvastatin sensitizes GBM cells to TMZ-induced cell death via a mechanism that involves autophagy and UPR pathways. More specifically, our results imply that the IRE1 and PERK signaling arms of the UPR regulate Simva-TMZ-mediated autophagy flux inhibition in U251 and U87 GBM cells. 

Retracted: No

PubPeer comments:
 1. The GAPDH for U251 in Fig. 1 has been reported as control for Fig. 4A. U87cells. 2. Again, the GAPDH control in Fig. 1, while disparately labeled for the lanes, used for Fig. 6A and 9A. 3. The protein loading controls (GAPDH) are the same for two different cells in Fig. 7, as well. ![file] ,