title: The thiazole derivative CPTH6 impairs autophagy.
abstract: We have previously demonstrated that the thiazole derivative 3-methylcyclopentylidene-[4-(4'-chlorophenyl)thiazol-2-yl]hydrazone (CPTH6) induces apoptosis and cell cycle arrest in human leukemia cells. The aim of this study was to evaluate whether CPTH6 is able to affect autophagy. By using several human tumor cell lines with different origins we demonstrated that CPTH6 treatment induced, in a dose-dependent manner, a significant increase in autophagic features, as imaged by electron microscopy, immunoblotting analysis of membrane-bound form of microtubule-associated protein 1 light chain 3 (LC3B-II) levels and by appearance of typical LC3B-II-associated autophagosomal puncta. To gain insights into the molecular mechanisms of elevated markers of autophagy induced by CPTH6 treatment, we silenced the expression of several proteins acting at different steps of autophagy. We found that the effect of CPTH6 on autophagy developed through a noncanonical mechanism that did not require beclin-1-dependent nucleation, but involved Atg-7-mediated elongation of autophagosomal membranes. Strikingly, a combined treatment of CPTH6 with late-stage autophagy inhibitors, such as chloroquine and bafilomycin A1, demonstrates that under basal condition CPTH6 reduces autophagosome turnover through an impairment of their degradation pathway, rather than enhancing autophagosome formation, as confirmed by immunofluorescence experiments. According to these results, CPTH6-induced enhancement of autophagy substrate p62 and NBR1 protein levels confirms a blockage of autophagic cargo degradation. In addition, CPTH6 inhibited autophagosome maturation and compounds having high structural similarities with CPTH6 produced similar effects on the autophagic pathway. Finally, the evidence that CPTH6 treatment decreased α-tubulin acetylation and failed to increase autophagic markers in cells in which acetyltransferase ATAT1 expression was silenced indicates a possible role of α-tubulin acetylation in CPTH6-induced alteration in autophagy. Overall, CPTH6 could be a valuable agent for the treatment of cancer and should be further studied as a possible antineoplastic agent.
authors.affiliation: Experimental Chemotherapy Laboratory, Regina Elena National Cancer Institute, Rome, Italy.
authors.forename: Y
authors.initials: Y
authors.lastname: Ragazzoni
authors.forename: M
authors.initials: M
authors.lastname: Desideri
authors.forename: C
authors.initials: C
authors.lastname: Gabellini
authors.forename: T
authors.initials: T
authors.lastname: De Luca
authors.forename: S
authors.initials: S
authors.lastname: Carradori
authors.forename: D
authors.initials: D
authors.lastname: Secci
authors.forename: R
authors.initials: R
authors.lastname: Nescatelli
authors.forename: A
authors.initials: A
authors.lastname: Candiloro
authors.forename: M
authors.initials: M
authors.lastname: Condello
authors.forename: S
authors.initials: S
authors.lastname: Meschini
authors.forename: D
authors.initials: D
authors.lastname: Del Bufalo
authors.forename: D
authors.initials: D
authors.lastname: Trisciuoglio
mesh_terms: D000123:Acetyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D001343:Autophagy / Q000187:drug effects*; D000071193:Autophagy-Related Protein 7; D045744:Cell Line, Tumor; D018922:HL-60 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D000071456:Sequestosome-1 Protein; D013844:Thiazoles / Q000737:chemistry / Q000494:pharmacology*; D044764:Ubiquitin-Activating Enzymes / Q000378:metabolism
chemical_list: C573972:3-methylcyclopentylidene-(4-(4'-chlorophenyl)thiazol-2-yl)hydrazone; D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents; D047908:Intracellular Signaling Peptides and Proteins; C477012:MAP1LC3A protein, human; D008869:Microtubule-Associated Proteins; C549756:NBR1 protein, human; D011506:Proteins; D034741:RNA, Small Interfering; C099718:SQSTM1 protein, human; D000071456:Sequestosome-1 Protein; D013844:Thiazoles; D000123:Acetyltransferases; C509349:ATG7 protein, human; D000071193:Autophagy-Related Protein 7; D044764:Ubiquitin-Activating Enzymes