Violacein induces death of resistant leukaemia cells via kinome reprogramming, endoplasmic reticulum stress and Golgi apparatus collapse.
It is now generally recognised that different modes of programmed cell death (PCD) are intimately linked to the cancerous process. However, the mechanism of PCD involved in cancer chemoprevention is much less clear and may be different between types of chemopreventive agents and tumour cell types involved. Therefore, from a pharmacological view, it is crucial during the earlier steps of drug development to define the cellular specificity of the candidate as well as its capacity to bypass dysfunctional tumoral signalling pathways providing insensitivity to death stimuli. Studying the cytotoxic effects of violacein, an antibiotic dihydro-indolone synthesised by an Amazon river Chromobacterium, we observed that death induced in CD34(+)/c-Kit(+)/P-glycoprotein(+)/MRP1(+) TF1 leukaemia progenitor cells is not mediated by apoptosis and/or autophagy, since biomarkers of both types of cell death were not significantly affected by this compound. To clarify the working mechanism of violacein, we performed kinome profiling using peptide arrays to yield comprehensive descriptions of cellular kinase activities. Pro-death activity of violacein is actually carried out by inhibition of calpain and DAPK1 and activation of PKA, AKT and PDK, followed by structural changes caused by endoplasmic reticulum stress and Golgi apparatus collapse, leading to cellular demise. Our results demonstrate that violacein induces kinome reprogramming, overcoming death signaling dysfunctions of intrinsically resistant human leukaemia cells.
Center for Experimental and Molecular Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Karla C S
KC
Queiroz
Renato
R
Milani
Roberta R
RR
Ruela-de-Sousa
Gwenny M
GM
Fuhler
Giselle Z
GZ
Justo
Willian F
WF
Zambuzzi
Nelson
N
Duran
Sander H
SH
Diks
C Arnold
CA
Spek
Carmen V
CV
Ferreira
Maikel P
MP
Peppelenbosch
D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000037:antagonists & inhibitors; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D002154:Calpain / Q000037:antagonists & inhibitors; D016923:Cell Death / Q000187:drug effects*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D064550:Death-Associated Protein Kinases; D059865:Endoplasmic Reticulum Stress; D006056:Golgi Apparatus / Q000187:drug effects*; D006801:Humans; D007211:Indoles / Q000627:therapeutic use*; D007938:Leukemia / Q000188:drug therapy*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism
D051017:Apoptosis Regulatory Proteins; D007211:Indoles; C579096:DAPK1 protein, human; D064550:Death-Associated Protein Kinases; D051057:Proto-Oncogene Proteins c-akt; D017868:Cyclic AMP-Dependent Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002154:Calpain; C063155:violacein