Dependence of ABCB1 transporter expression and function on distinct sphingolipids generated by ceramide synthases-2 and -6 in chemoresistant renal cancer
| Autor: | Wing-Kee Lee, Michelle Maaß, Amy Quach, Nataliya Poscic, Holly Prangley, Erin-Claire Pallott, Jiyoon L. Kim, Jason S. Pierce, Besim Ogretmen, Anthony H. Futerman, Frank Thévenod |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
TG
thapsigargin Male cancer drug resistance FASN fatty acid synthase Biochemistry ERAD ER-associated degradation UPR unfolded protein response MDR multidrug resistance Tandem Mass Spectrometry Sphingosine N-Acyltransferase Tumor Microenvironment HERPUD1 homocysteine-inducible ER protein with ubiquitin-like domain 1 GRP78 glucose-regulated protein 78 kDa unfolded protein response Endoplasmic Reticulum-Associated Degradation Kidney Neoplasms XBP1 X-box binding protein 1 lipids (amino acids peptides and proteins) Female ABC transporter ATP6α activating transcription factor 6α Research Article ATP Binding Cassette Transporter Subfamily B PDIA4 protein disulfide isomerase family A member 4 P-glycoprotein CHOP C/EBP homologous protein Ceramides ACACA acetyl-CoA carboxylase alpha ER endoplasmic reticulum VLC very long chain LCC long-chain ceramide HPCT human proximal convoluted tubule Humans RNA Messenger TUN tunicamycin Molecular Biology Sphingolipids SM sphingomyelin UCSC University of California Santa Cruz SREBP sterol-regulatory element binding protein LD lipid domain Tumor Suppressor Proteins Membrane Proteins Cell Biology DOX doxorubicin Doxorubicin Drug Resistance Neoplasm Mff mitochondrial fission factor CerS ceramide synthase HMGCL 3-hydroxymethyl-3-methylglutaryl-CoA lyase TM transmembrane |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | Oncogenic multidrug resistance (MDR) is commonly intrinsic to renal cancer based on the physiological expression of detoxification transporters, particularly ABCB1, thus hampering chemotherapy. ABCB1 activity is directly dependent on its lipid microenvironment, localizing to cholesterol- and sphingomyelin-rich domains. As ceramides are the sole source for sphingomyelins, we hypothesized that ceramide synthase (CerS)-derived ceramides regulate ABCB1 activity. Using data from RNA-seq databases, we found patient kidney tumors exhibited increased CerS2 mRNA, which was inversely correlated with CerS6 mRNA in ABCB1+ clear cell carcinomas. Endogenous elevated CerS2 and lower CerS5/6 mRNA and protein resulted in disproportionately higher CerS2 to CerS5/6 activities (2-fold) in chemoresistant ABCB1high (A498, Caki-1) compared to chemosensitive ABCB1low (ACHN, HPCT) cells. In addition, lipidomics analyses by HPLC-MS/MS showed bias towards CerS2-associated C20:0/C20:1-ceramides compared to CerS5/6-associated C14:0/C16:0-ceramides (2:1). Sphingomyelins were similarly altered. We demonstrated that chemoresistance to doxorubicin in ABCB1high cells was partially reversed by inhibitors of de novo ceramide synthesis (L-cycloserine) and CerS (fumonisin B1) in cell viability assays. Additionally, downregulation of CerS2/6, but not CerS5, attenuated ABCB1 mRNA, protein, plasma membrane localization, rhodamine 123+ efflux transport activity, and doxorubicin resistance. Similar findings were observed with catalytically-inactive CerS6-H212A. Furthermore, CerS6-targeting siRNA shifted ceramide and sphingomyelin composition to ultra long-chain species (C22-C26). Inhibitors of ER-associated degradation (ERAD) (eeyarestatin I) and the proteasome (MG132, bortezomib) prevented ABCB1 loss induced by CerS2/6 downregulation. We conclude that a critical balance in ceramide/sphingomyelin species is prerequisite to ABCB1 expression and functionalization, which could be targeted to reverse MDR in renal cancers. Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved. |
| Databáze: | OpenAIRE |
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