PMID- 37419396 OWN - NLM STAT- MEDLINE DCOM- 20230807 LR - 20230807 IS - 1879-260X (Electronic) IS - 0925-4439 (Linking) VI - 1869 IP - 7 DP - 2023 Oct TI - Melanoma cells with acquired resistance to vemurafenib have decreased autophagic flux and display enhanced ability to transfer resistance. PG - 166801 LID - S0925-4439(23)00167-9 [pii] LID - 10.1016/j.bbadis.2023.166801 [doi] AB - Over the last years, the incidence of melanoma, the deadliest form of skin cancer, has risen significantly. Nearly half of the melanoma patients exhibit the BRAF(V600E) mutation. Although the use of BRAF and MEK inhibitors (BRAFi and MEKi) showed an impressive success rate in melanoma patients, durability of response remains an issue because tumor quickly becomes resistant. Here, we generated and characterized Lu1205 and A375 melanoma cells resistant to vemurafenib (BRAFi). Resistant cells (Lu1205R and A375R) exhibit higher IC(50) (5-6 fold increase) and phospho-ERK levels and 2-3 times reduced apoptosis than their sensitive parents (Lu1205S and A375S). Moreover, resistant cells are 2-3 times bigger, display a more elongated morphology and have a modulation of migration capacity. Interestingly, pharmacological inhibition of sphingosine kinases, that prevents sphingosine-1-phosphate production, reduces migration of Lu1205R cells by 50 %. In addition, although Lu1205R cells showed increased basal levels of the autophagy markers LC3II and p62, they have decreased autophagosome degradation and autophagy flux. Remarkably, expression of Rab27A and Rab27B, which are involved in the release of extracellular vesicles are dramatically augmented in resistant cells (i.e. 5-7 fold increase). Indeed, conditioned media obtained from Lu1205R cells increased the resistance to vemurafenib of sensitive cells. Hence, these results support that resistance to vemurafenib modulates migration and the autophagic flux and may be transferred to nearby sensitive melanoma cells by factors that are released to the extracellular milieu by resistant cells. CI - Copyright (c) 2023 Elsevier B.V. All rights reserved. FAU - Perez, Celia N AU - Perez CN AD - Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis (UNSL), Ejercito de los Andes 950, 5700 San Luis, Argentina; Instituto Multidisciplinario de Investigaciones Biologicas (IMIBIO-SL), CONICET, Argentina. FAU - Falcon, Cristian R AU - Falcon CR AD - Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis (UNSL), Ejercito de los Andes 950, 5700 San Luis, Argentina; Instituto Multidisciplinario de Investigaciones Biologicas (IMIBIO-SL), CONICET, Argentina. FAU - Mons, Johinna Delgado AU - Mons JD AD - Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis (UNSL), Ejercito de los Andes 950, 5700 San Luis, Argentina; Instituto Multidisciplinario de Investigaciones Biologicas (IMIBIO-SL), CONICET, Argentina. FAU - Orlandi, Federico Cuello AU - Orlandi FC AD - Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis (UNSL), Ejercito de los Andes 950, 5700 San Luis, Argentina; Instituto Multidisciplinario de Investigaciones Biologicas (IMIBIO-SL), CONICET, Argentina. FAU - Sangiacomo, Mercedes AU - Sangiacomo M AD - Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis (UNSL), Ejercito de los Andes 950, 5700 San Luis, Argentina; Instituto Multidisciplinario de Investigaciones Biologicas (IMIBIO-SL), CONICET, Argentina. FAU - Fernandez-Munoz, Juan M AU - Fernandez-Munoz JM AD - Instituto de Biologia y Medicina Experimental de Cuyo (IMBECU), CONICET, Argentina. FAU - Guerrero, Martin AU - Guerrero M AD - Instituto de Biologia y Medicina Experimental de Cuyo (IMBECU), CONICET, Argentina. FAU - Benito, Paula G AU - Benito PG AD - Instituto de Histologia y Embriologia de Mendoza (IHEM), Universidad Nacional de Cuyo-CONICET, Argentina. FAU - Colombo, Maria I AU - Colombo MI AD - Instituto de Histologia y Embriologia de Mendoza (IHEM), Universidad Nacional de Cuyo-CONICET, Argentina. FAU - Zoppino, Felipe C M AU - Zoppino FCM AD - Instituto de Biologia y Medicina Experimental de Cuyo (IMBECU), CONICET, Argentina. FAU - Alvarez, Sergio E AU - Alvarez SE AD - Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis (UNSL), Ejercito de los Andes 950, 5700 San Luis, Argentina; Instituto Multidisciplinario de Investigaciones Biologicas (IMIBIO-SL), CONICET, Argentina. Electronic address: sealvar@email.unsl.edu.ar. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20230706 PL - Netherlands TA - Biochim Biophys Acta Mol Basis Dis JT - Biochimica et biophysica acta. Molecular basis of disease JID - 101731730 RN - 207SMY3FQT (Vemurafenib) RN - EC 2.7.11.1 (Proto-Oncogene Proteins B-raf) RN - 0 (Sulfonamides) RN - 0 (Indoles) RN - 0 (Protein Kinase Inhibitors) SB - IM MH - Humans MH - Vemurafenib/pharmacology MH - *Proto-Oncogene Proteins B-raf/genetics/metabolism MH - Sulfonamides/pharmacology MH - Indoles/pharmacology MH - Drug Resistance, Neoplasm/genetics MH - Cell Line, Tumor MH - Xenograft Model Antitumor Assays MH - *Melanoma/drug therapy/genetics/metabolism MH - Protein Kinase Inhibitors/pharmacology MH - Autophagy OTO - NOTNLM OT - Autophagy OT - BRAF(V600E) mutation OT - Melanoma OT - Resistance transfer OT - Vemurafenib COIS- Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2023/07/08 10:42 MHDA- 2023/08/07 06:41 CRDT- 2023/07/07 19:27 PHST- 2022/12/27 00:00 [received] PHST- 2023/05/31 00:00 [revised] PHST- 2023/06/28 00:00 [accepted] PHST- 2023/08/07 06:41 [medline] PHST- 2023/07/08 10:42 [pubmed] PHST- 2023/07/07 19:27 [entrez] AID - S0925-4439(23)00167-9 [pii] AID - 10.1016/j.bbadis.2023.166801 [doi] PST - ppublish SO - Biochim Biophys Acta Mol Basis Dis. 2023 Oct;1869(7):166801. doi: 10.1016/j.bbadis.2023.166801. Epub 2023 Jul 6.