title: A flavonoid isolated from Streptomyces sp. (ERINLG-4) induces apoptosis in human lung cancer A549 cells through p53 and cytochrome c release caspase dependant pathway.
abstract: The aim of this study was to investigate the anticancer activity of a flavonoid type of compound isolated from soil derived filamentous bacterium Streptomyces sp. (ERINLG-4) and to explore the molecular mechanisms of action. Cytotoxic properties of ethyl acetate extract was carried out against A549 lung cancer cell line using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cytotoxic properties of isolated compound were investigated in A549 lung cancer cell line, COLO320DM cancer cell line and Vero cells. The compound showed potent cytotoxic properties against A549 lung cancer cell line and moderate cytotoxic properties against COLO320DM cancer cell line. Isolated compound showed no toxicity up to 2000 μg/mL in Vero cells. So we have chosen the A549 lung cancer cell line for further anticancer studies. Intracellular visualization was done by using a laser scanning confocal microscope. Apoptosis was measured using DNA fragmentation technique. Treatment of the A549 cancer cells with isolated compound significantly reduced cell proliferation, increased formation of fragmented DNA and apoptotic body. Activation of caspase-9 and caspase-3 indicated that compound may be inducing intrinsic and extrinsic apoptosis pathways. Bcl-2, p53, pro-caspases, caspase-3, caspase-9 and cytochrome c release were detected by western blotting analysis after compound treatment (123 and 164 μM). The activities of pro-caspases-3, caspase-9 cleaved to caspase-3 and caspase-9 gradually increased after the addition of isolated compound. But Bcl-2 protein was down regulated after treatment with isolated compound. Molecular docking studies showed that the compound bound stably to the active sites of caspase-3 and caspase-9. These results strongly suggest that the isolated compound induces apoptosis in A549 cancer cells via caspase activation through cytochrome c release from mitochondria. The present results might provide helpful suggestions for the design of antitumor drugs toward lung cancer treatment.
authors.affiliation: Division of Microbiology and Cancer Biology, Entomology Research Institute, Loyola College, Chennai 600 034, India.
authors.forename: C
authors.initials: C
authors.lastname: Balachandran
authors.affiliation: Department of Toxicology, Advinus Therapeutics Ltd, Bangalore 560058, India.
authors.forename: B
authors.initials: B
authors.lastname: Sangeetha
authors.affiliation: Division of Microbiology and Cancer Biology, Entomology Research Institute, Loyola College, Chennai 600 034, India; Department of Botany and Microbiology, College of Science, King Saud University, P.O.Box. 2455, Riyadh 11451, Saudi Arabia. Electronic address: avdpandiyan@yahoo.co.in.
authors.forename: V
authors.initials: V
authors.lastname: Duraipandiyan
authors.affiliation: Research and Development Centre, Orchid Chemicals and Pharmaceuticals Ltd, Sozhanganallur, Chennai 600119, India.
authors.forename: M Karunai
authors.initials: MK
authors.lastname: Raj
authors.affiliation: Division of Microbiology and Cancer Biology, Entomology Research Institute, Loyola College, Chennai 600 034, India.
authors.forename: S
authors.initials: S
authors.lastname: Ignacimuthu
authors.affiliation: Department of Botany and Microbiology, College of Science, King Saud University, P.O.Box. 2455, Riyadh 11451, Saudi Arabia.
authors.forename: N A
authors.initials: NA
authors.lastname: Al-Dhabi
authors.affiliation: Division of Microbiology and Cancer Biology, Entomology Research Institute, Loyola College, Chennai 600 034, India.
authors.forename: K
authors.initials: K
authors.lastname: Balakrishna
authors.affiliation: Research and Development Centre, Orchid Chemicals and Pharmaceuticals Ltd, Sozhanganallur, Chennai 600119, India.
authors.forename: K
authors.initials: K
authors.lastname: Parthasarathy
authors.affiliation: Department of Bioinformatics, Orchid Chemicals and Pharmaceuticals Ltd, Research and Development Centre, Shozanganallur, Chennai 600 119, India.
authors.forename: N M
authors.initials: NM
authors.lastname: Arulmozhi
authors.affiliation: Department of Botany and Microbiology, College of Science, King Saud University, P.O.Box. 2455, Riyadh 11451, Saudi Arabia.
authors.forename: M Valan
authors.initials: MV
authors.lastname: Arasu
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002522:Chlorocebus aethiops; D045304:Cytochromes c / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D005419:Flavonoids / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology*; D013302:Streptomyces / Q000737:chemistry*; D013329:Structure-Activity Relationship; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D014709:Vero Cells
chemical_list: D000970:Antineoplastic Agents; D005419:Flavonoids; D016159:Tumor Suppressor Protein p53; D045304:Cytochromes c; D020169:Caspases
keywords: Caspases; Cytochrome c; Molecular docking; Streptomyces sp.; p53