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cdk6 cyclin d1  (SignalChem)
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Effect of aspirin, salicylic acid, 2,3-DHBA, 2,6-DHBA and 2,4,6-THBA (at 0.5 mM) on CDK2, 4 and 6 enzyme activity. (A) Upper panel represents in vitro kinase assays showing the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA on CDK2 activity. Lower panel shows coomassie stained H1 histones. (B) Quantification of the blot in (A). (C) Upper panel represents in vitro kinase assays showing the dose-dependent effect of 2,4,6-THBA on CDK2 activity. Lower panel shows coomassie stained H1 histones. (D) Quantification of the blot in (C). (E) Upper panel represents the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA and 2,4,6-THBA on CDK4 activity. Lower panel shows coomassie stained retinoblastoma protein. (F) Quantification of the blot in (E). (G) Upper panel represents the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA (0.5 mM) and 2,4,6-THBA on <t>CDK6</t> enzyme activity. Lower panel shows coomassie stained retinoblastoma protein. (H) Quantification of the blot in (G). * P<0.05, ** P<0.01, † P<0.001, ‡ P<0.001.
Cdk6 Cyclin D1, supplied by SignalChem, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Effect of aspirin, salicylic acid, 2,3-DHBA, 2,6-DHBA and 2,4,6-THBA (at 0.5 mM) on CDK2, 4 and 6 enzyme activity. (A) Upper panel represents in vitro kinase assays showing the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA on CDK2 activity. Lower panel shows coomassie stained H1 histones. (B) Quantification of the blot in (A). (C) Upper panel represents in vitro kinase assays showing the dose-dependent effect of 2,4,6-THBA on CDK2 activity. Lower panel shows coomassie stained H1 histones. (D) Quantification of the blot in (C). (E) Upper panel represents the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA and 2,4,6-THBA on CDK4 activity. Lower panel shows coomassie stained retinoblastoma protein. (F) Quantification of the blot in (E). (G) Upper panel represents the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA (0.5 mM) and 2,4,6-THBA on CDK6 enzyme activity. Lower panel shows coomassie stained retinoblastoma protein. (H) Quantification of the blot in (G). * P<0.05, ** P<0.01, † P<0.001, ‡ P<0.001.

Journal: International Journal of Oncology

Article Title: Salicylic acid metabolites and derivatives inhibit CDK activity: Novel insights into aspirin's chemopreventive effects against colorectal cancer

doi: 10.3892/ijo.2017.4167

Figure Lengend Snippet: Effect of aspirin, salicylic acid, 2,3-DHBA, 2,6-DHBA and 2,4,6-THBA (at 0.5 mM) on CDK2, 4 and 6 enzyme activity. (A) Upper panel represents in vitro kinase assays showing the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA on CDK2 activity. Lower panel shows coomassie stained H1 histones. (B) Quantification of the blot in (A). (C) Upper panel represents in vitro kinase assays showing the dose-dependent effect of 2,4,6-THBA on CDK2 activity. Lower panel shows coomassie stained H1 histones. (D) Quantification of the blot in (C). (E) Upper panel represents the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA and 2,4,6-THBA on CDK4 activity. Lower panel shows coomassie stained retinoblastoma protein. (F) Quantification of the blot in (E). (G) Upper panel represents the effect of aspirin, salicylic acid, 2,3-DHBA and 2,6-DHBA (0.5 mM) and 2,4,6-THBA on CDK6 enzyme activity. Lower panel shows coomassie stained retinoblastoma protein. (H) Quantification of the blot in (G). * P<0.05, ** P<0.01, † P<0.001, ‡ P<0.001.

Article Snippet: CDK1/cyclin B1 active enzyme from New England Biolabs (NEB) (Ipswich, MA, USA), CDK1/cyclin B1, CDK2/cyclin A2, CDK4/cyclin D1, CDK6/cyclin D1, Retinoblastoma (C-term) and kinase buffer were purchased from SignalChem (Richmond, BC, Canada).

Techniques: Activity Assay, In Vitro, Staining

A model depicting how aspirin may preferentially act on colonic tissue to protect against CRC. We suggest that unabsorbed aspirin/salicylic acid in the stomach and upper intestine is passed on to the colon, taken up by the colonic epithelial cells, and metabolized by CYP450s to produce 2,3-DHBA and 2,5-DHBAs. These hydrophilic DHBAs may not easily cross the basolateral membrane of the epithelial cells and within these cells, they may accumulate to pharmacologically relevant concentrations, leading to the inhibition of CDK1 and CDK6, and exert anticancer effects. Colonic epithelial cells may also get exposed to salicylic acid metabolites generated by the gut microflora, and uptake of these HBAs by cells may also cause CDK inhibition. This additional layer of protection against tumor development may be unique to the colonic tissue allowing aspirin as a more effective drug against CRC. (?), Aspirin acetylates CDK1; however, how this modification affects its function is not clear at this stage.

Journal: International Journal of Oncology

Article Title: Salicylic acid metabolites and derivatives inhibit CDK activity: Novel insights into aspirin's chemopreventive effects against colorectal cancer

doi: 10.3892/ijo.2017.4167

Figure Lengend Snippet: A model depicting how aspirin may preferentially act on colonic tissue to protect against CRC. We suggest that unabsorbed aspirin/salicylic acid in the stomach and upper intestine is passed on to the colon, taken up by the colonic epithelial cells, and metabolized by CYP450s to produce 2,3-DHBA and 2,5-DHBAs. These hydrophilic DHBAs may not easily cross the basolateral membrane of the epithelial cells and within these cells, they may accumulate to pharmacologically relevant concentrations, leading to the inhibition of CDK1 and CDK6, and exert anticancer effects. Colonic epithelial cells may also get exposed to salicylic acid metabolites generated by the gut microflora, and uptake of these HBAs by cells may also cause CDK inhibition. This additional layer of protection against tumor development may be unique to the colonic tissue allowing aspirin as a more effective drug against CRC. (?), Aspirin acetylates CDK1; however, how this modification affects its function is not clear at this stage.

Article Snippet: CDK1/cyclin B1 active enzyme from New England Biolabs (NEB) (Ipswich, MA, USA), CDK1/cyclin B1, CDK2/cyclin A2, CDK4/cyclin D1, CDK6/cyclin D1, Retinoblastoma (C-term) and kinase buffer were purchased from SignalChem (Richmond, BC, Canada).

Techniques: Inhibition, Generated, Modification