Elucidation of the molecular mechanisms of a salicylhydrazide class of compounds by proteomic analysis.
Previously, we described a series of salicylhydrazide compounds with potent anti-cancer activities against a panel of human cancer cell lines derived from different origins. Preclinical evaluation showing efficacy both in vitro and in vivo in human cancer models indicated that these agents may represent a promising class of anticancer drugs. In the present study, we performed an in-depth investigation on the underlying molecular mechanisms of the most potent compounds, SC21 and SC23, using a proteomic method and bioinformatics tools. We demonstrated that SC23 induced apoptosis through multiple signaling pathways. In particular, SC23 regulated the expression of Bcl-2, p21, acetylated histone H3 and beta-tubulin and the combined modulation of these proteins may result in the induction of apoptosis. We also examined the effect of SC21 and SC23 on cell cycle progression and found that both compounds arrested cells in S-phase in most cell lines tested. To better understand the signaling networks involved, we analyzed the SC21- and SC23-treated cell lysates by the Kinexus 628 antibody microarray. The results were interpreted with the aid of Ingenuity Pathway Analysis (IPA) software. It was found that SC21 interfered with JAK/STAT signaling and elicited apoptosis through Fas and caspases pathways. Unlike SC21, SC23 induced RAR activation and caused cell cycle arrest. The signaling networks identified by this work may provide the basis for future mechanistic studies. The validation of the proposed pathways and the elucidation of the signaling cross-talk are currently under way.
Cao
Xuefei
X
Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA 90089, USA.
Plasencia
Carmen
C
Kanzaki
Atsuko
A
Yang
Austin
A
Burke
Terrence R
TR
Neamati
Nouri
N
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival; D053938:DNA Fragmentation / Q000187:drug effects; D030541:Databases, Genetic; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D006834:Hydrazines / Q000138:chemical synthesis / Q000494:pharmacology*; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism; D040901:Proteomics; D017382:Reactive Oxygen Species / Q000378:metabolism; D018168:Receptors, Retinoic Acid / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D053719:Tandem Mass Spectrometry
D000970:Antineoplastic Agents; D006834:Hydrazines; D017382:Reactive Oxygen Species; D018168:Receptors, Retinoic Acid; C000687:salicyl hydrazide