Protein kinase inhibitors have been widely used as therapeutic agents to treat a variety of diseases, but many of them may cause off-target effects by unexpectedly targeting other noncognate kinases due to high conversion across the protein kinase family. The mitogen-activated protein kinase (MAPK) signaling pathway plays an essential role in tumorigenesis, which has been recognized as a high priority in the druggable target candidates of anticancer therapy. Here, we attempt to investigate the untargeted kinase-inhibitor interactions (UKIIs) of kinase-targeted therapies for the cancer MAPK signaling cascade via an integration of biomolecular modeling, cell viability assay and kinase inhibition analysis. A systematic kinase-inhibitor interaction profile is created for 28 FDA-approved kinase inhibitor drugs across 9 caner-related MAPK kinases. The created profile is analyzed at structural, energetic and dynamic levels and, consequently, totally 18 promising UKII pairs with high theoretical affinity are derived, from which the noncognate inhibitors Cabozantinib, Regorafenib and Crizotinib are selected to test their cytotoxic effects on human epithelial colorectal adenocarcinoma Caco-2 cell line and inhibition activity against the recombinant protein of human p38α kinase domain. The obtained results are compared with two cognate MAPK inhibitors JNK-IN-8 and BIRB796. As might be expected, the Regorafenib, Crizotinib and Cabozantinib exhibit high, moderate and low cytotoxicities, respectively. In addition, the Regorafenib is determined to have a potent p38α-inhibitory activity. This is basically in line with the test results of positive controls JNK-IN-8 and BIRB796 and can be well confirmed by computational modeling.

OBJECTIVE: To investigate the possible biological mechanism of dental fluorosis at a molecular level.
METHODS: Cultured LS8 were incubated with serum-free medium containing selected concentrations of NaF (0 ∼ 2 mM) for either 24 or 48 h. Subcellular microanatomy was characterized using TEM; meanwhile, selected biomolecules were analysed using various biochemistry techniques. Transient transfection was used to modulate a molecular pathway for apoptosis.
RESULTS: Apoptosis of LS8 was induced by NaF treatment that showed both time and concentration dependency. The activity of caspase-3, -8, -9 was found to be increased with NaF in a dose-dependent manner. Western blot revealed that the protein expression of p-ERK and p-JNK were decreased, while the expression of p-P38 was increased. Inhibition of the p-ERK and p-JNK pathways resulted in a similar decrease for caspase-3.
CONCLUSIONS: During NaF-induced apoptosis of LS8, p-ERK and p-JNK were closely associated with induction of apoptosis, which might be a mechanism of dental fluorosis.

This study sought to determine the serum levels of chemerin in gastric cancer patients and healthy subjects and to investigate the biological effect of chemerin on gastric cancer cells. Serum chemerin level of 36 gastric cancer patients and 40 healthy subjects was measured by enzyme-linked immunosorbent assay. AGS and MKN28 cells were treated with recombinant human chemerin, MAPKs phosphorylation was then measured. Chemerin were added to culture medium of AGS and MKN28 in the absence or presence of MAPK inhibitors, VEGF, MMP-7, IL-6 and cell invasiveness assay were then performed. Serum level of chemerin was significantly higher in gastric cancer patients than healthy subjects (P<0.01). The elevation of serum chemerin level was associated with advanced clinical stages and nonintestinal type of gastric cancer. Chemerin increased invasiveness of gastric cancer cells. Chemerin induced phosphorylation of p38 and ERK1/2 MAPKs and upregulated VEGF, MMP-7 and IL-6. Inhibition of ERK1/2 phosphorylation abolished the upregulation of VEGF, MMP-7 and IL-6 and the pro-invasive effect of chemerin. This study demonstrates a novel action of chemerin in gastric carcinogenesis.

Furan is a chemical hepatocarcinogen in mice and rats. Its previously postulated cancer mode of action (MOA) is chronic cytotoxicity followed by sustained regenerative proliferation; however, its molecular basis is unknown. To this end, we conducted toxicogenomic analysis of B3C6F1 mouse livers following three week exposures to non-carcinogenic (0, 1, 2mg/kgbw) or carcinogenic (4 and 8mg/kgbw) doses of furan. We saw enrichment for pathways responsible for cytotoxicity: stress-activated protein kinase (SAPK) and death receptor (DR5 and TNF-alpha) signaling, and proliferation: extracellular signal-regulated kinases (ERKs) and TNF-alpha. We also noted the involvement of NF-kappaB and c-Jun in response to furan, which are genes that are known to be required for liver regeneration. Furan metabolism by CYP2E1 produces cis-2-butene-1,4-dial (BDA), which is required for ensuing cytotoxicity and oxidative stress. NRF2 is a master regulator of gene expression during oxidative stress and we suggest that chronic NFR2 activity and chronic inflammation may represent critical transition events between the adaptive (regeneration) and adverse (cancer) outcomes. Another objective of this study was to demonstrate the applicability of toxicogenomics data in quantitative risk assessment. We modeled benchmark doses for our transcriptional data and previously published cancer data, and observed consistency between the two. Margin of exposure values for both transcriptional and cancer endpoints were also similar. In conclusion, using furan as a case study we have demonstrated the value of toxicogenomics data in elucidating dose-dependent MOA transitions and in quantitative risk assessment.