Multifactorial Modes of Action of Arsenic Trioxide in Cancer Cells as Analyzed by Classical and Network Pharmacology

Abstract

Arsenic trioxide is a traditional remedy in Chinese Medicine since ages. Nowadays, it is clinically used to treat acute promyelocytic leukemia (APL) by targeting PML/RARA. However, the drug’s activity is broader and the mechanisms of action in other tumor types remain unclear. In this study, we investigated molecular modes of action by classical and network pharmacological approaches. CEM/ADR5000 resistance leukemic cells were similar sensitive to As2O3 as their wild-type counterpart CCRF-CEM (resistance ratio: 1.88). Drug-resistant U87.MG ΔEGFR glioblastoma cells harboring mutated epidermal growth factor receptor were even more sensitive (collateral sensitive) than wild-type U87.MG cells (resistance ratio: 0.33). HCT-116 colon carcinoma p53-/- knockout cells were 7.16-fold resistant toward As2O3 compared to wild-type cells. Forty genes determining cellular responsiveness to As2O3 were identified by microarray and COMPARE analyses in 58 cell lines of the NCI panel. Hierarchical cluster analysis-based heat mapping revealed significant differences between As2O3 sensitive cell lines and resistant cell lines with p-value: 1.86 × 10-5. The genes were subjected to Galaxy Cistrome gene promoter transcription factor analysis to predict the binding of transcription factors. We have exemplarily chosen NF-kB and AP-1, and indeed As2O3 dose-dependently inhibited the promoter activity of these two transcription factors in reporter cell lines. Furthermore, the genes identified here and those published in the literature were assembled and subjected to Ingenuity Pathway Analysis for comprehensive network pharmacological approaches that included all known factors of resistance of tumor cells to As2O3. In addition to pathways related to the anticancer effects of As2O3, several neurological pathways were identified. As arsenic is well-known to exert neurotoxicity, these pathways might account for neurological side effects. In conclusion, the activity of As2O3 is not restricted to acute promyelocytic leukemia. In addition to PML/RARA, numerous other genes belonging to diverse functional classes may also contribute to its cytotoxicity. Network pharmacology is suited to unravel the multifactorial modes of action of As2O3.

Collateral Sensitivity of Parthenolide via NF-κB and HIF-α Inhibition and Epigenetic Changes in Drug-Resistant Cancer Cell Lines

“…Importantly, COMPARE analyses and Cluster models have been previously validated for gene expression profiling and for approaching molecular pharmacology of anti-tumor compounds. This method was previously described by us in detail (Dawood et al, 2018).…”

“…Prior to the analysis, genes were uploaded to IPA in Excel format. Core analyses were carried out to identify canonical pathways, diseases and functions, and relevant networks as described (Dawood et al, 2018).…”

Repurposing of Bromocriptine for Cancer Therapy

“…Afterward, NF-κB activity was induced with 100 ng/mL of TNF-α for 24 h. The activation was measured by detecting SEAP spectrophotometrically at 630 nm with addition of Quanti Blue (Invivogen). The procedure has been reported by us ( Kadioglu and Efferth, 2015 ; Kadioglu et al, 2016 ; Dawood et al, 2018 ).…”

Identification of potential novel drug resistance mechanisms by genomic and transcriptomic profiling of colon cancer cells with p53 deletion

“…The drug resistance profile of HCT116 p53 −/− has been studied during the past years. Compared to wild-type cells, these knockout cells reveal resistance to established anticancer drugs of diverse pharmacological classes (doxorubicin, 5-fluorouracil and 5′-deoxy-5-fluorouridine, cisplatin and oxaliplatin, etoposide, and vincristine) as well as to investigational cytotoxic compounds with activity against cancer (arsenic trioxide as PML/RARA inhibitor, nutlin-3a as p53 activator, the fluoropyrimidine F10, the HDAC inhibitor entinostat and the synthetic polyamine DENSpm) and even cytotoxic but non-cancer drugs (the antimalarial quinacrine, the anticonvulsant valproic acid and the anti-inflammatory and COX1/2-inhibitory ibuprofen (Brachtendorf et al 2018 ; Bunz et al 1999 ; Coker-Gurkan et al 2015 ; Dawood et al 2018 ; Dominijanni and Gmeiner 2018 ; Gunasegaran et al 2020 ; Hernlund et al 2008 ; Janssen et al 2008 ; Kralova et al 2009 ; Lin et al 2004 ; Mohapatra et al 2012 ; Sonnemann et al 2014 ; Terranova-Barberio et al 2017 ).…”