title: Danazol has potential to cause PKC translocation, cell cycle dysregulation, and apoptosis in breast cancer cells.
abstract: Danazol, the established clinical drug, has given promising therapeutic results in a series of clinical trials with breast cancer patients. Danazol shares structural similarities with several known PKC agonists and fits well into the C1 domain. Danazol binds to the C1b domain of PKC with Kdof 5.64 ± 1.27 μm. MD simulation studies further support that the PKC-danazol molecular model is stable and showing minimum distortion to the structure during the simulation period. Immunofluorescence and Western blotting studies indicate that MDAMB-231 cells stimulated with danazol exhibit translocation of PKCα to the plasma membrane. Cells stimulated with danazol causes appearance of several phosphorylated proteins in lysate and plasma membrane. In addition, danazol affects carcinogenic molecule (PMA)-induced intracellular signaling in cancer cells. It halted the cancer cells in the G1 phase of the cell cycle and reduced the viability of ER+veand triple-negative breast cancer cells with an IC50of 31 ± 2.63 and 65 ± 4.27 μg/ml, respectively. DNA fragmentation and flow cytometry experiments revealed that the cell death follows the apoptotic pathway. It affects mitochondrial membrane potentials and releases cytochrome-C from mitochondria to induce downstream apoptosis in breast cancer cells. Hence, the current study may help clinicians to re-design their treatment strategy to optimize therapeutic potentials of the molecule.
authors.affiliation: Malaria Research Group, Department of Biosciences and Bioengineering, Indian Institute of Technology-Guwahati, Guwahati, Assam, India.
authors.forename: Suman Jyoti
authors.initials: SJ
authors.lastname: Deka
authors.affiliation: Department of Chemistry, Indian Institute of Technology-Guwahati, Guwahati, Assam, India.
authors.forename: Ashalata
authors.initials: A
authors.lastname: Roy
authors.affiliation: Molecular Informatics & Design Laboratory, Department of Biotechnology, Indian Institute of Technology-Guwahati, Guwahati, Assam, India.
authors.forename: Vibin
authors.initials: V
authors.lastname: Ramakrishnan
authors.affiliation: Department of Chemistry, Indian Institute of Technology-Guwahati, Guwahati, Assam, India.
authors.forename: Debasis
authors.initials: D
authors.lastname: Manna
authors.affiliation: Malaria Research Group, Department of Biosciences and Bioengineering, Indian Institute of Technology-Guwahati, Guwahati, Assam, India.
authors.forename: Vishal
authors.initials: V
authors.lastname: Trivedi
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000188:drug therapy*; D002453:Cell Cycle / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D003613:Danazol / Q000494:pharmacology* / Q000627:therapeutic use; D016503:Drug Delivery Systems; D005260:Female; D006801:Humans; D015151:Immunoblotting; D020128:Inhibitory Concentration 50; D061986:MCF-7 Cells; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D011493:Protein Kinase C; D003613:Danazol
keywords: chemical biology; cheminformatics; drug discovery; enzymatic mechanism; kinase; phosphatase