title: 3-cinnamoyl-4-hydroxy-6-methyl-2H-pyran-2-one (CHP) inhibits human ovarian cell proliferation by inducing apoptosis.
abstract: Coumarins induce apoptosis by activating mitochondrial pathway and caspase-3-dependent apoptotic pathway. In the present study, we first time investigated the effect of 3-cinnamoyl-4-hydroxy-6-methyl-2H-pyran-2-one (CHP) on induction of apoptosis in human ovarian carcinoma cells. The data from MTT assay revealed a significant inhibitory effect on cell viability at 30 (87%) and 50 μM (74%) concentration of CHP in OVCAR-3 and OVCAR-420 cells, respectively after 72 h. Apoptosis analysis using annexin V/PI double staining followed by flow cytometry showed 59 and 52% binding to annexin V-FITC in OVCAR-3 and OVCAR-420 cells respectively. propidium iodide (PI) staining and flow cytometry examination indicated a significant increase in percentage of cells in G2/M phase after treatment with CHP compared to DMSO control group. Reactive oxygen species (ROS) assay kit showed increase in levels of ROS. We used rhodamine-123 (Rh-123) staining and flow cytometry assay to determine changes in mitochondrial membrane potential (ΛΨm). The results revealed that CHP significantly decreased MMP to 85.65 ± 1.2443% & 49.78 ± 1.6554% at 10 and 30 μM respectively in OVCAR-3 compared to 95.97 ± 2.1243% in control group. Western blot analysis clearly indicated a significant increase in the expression of Caspase-3, Bax, and release of Cytochrome c and decrease in Bcl-2, CDK1 and Cyclin B1 expression on treatment with CHP. Therefore, CHP may become a potential candidate for the treatment of human ovarian cancers.
authors.affiliation: Department of Oncology, The Second Affiliated Hospital of Nanchang University Nanchang 330006, P. R. China.
authors.forename: Qiong-Yu
authors.initials: QY
authors.lastname: Lan
authors.affiliation: Department of Oncology, The Second Affiliated Hospital of Nanchang University Nanchang 330006, P. R. China.
authors.forename: Que-Ling
authors.initials: QL
authors.lastname: Liu
authors.affiliation: Department of Oncology, The Second Affiliated Hospital of Nanchang University Nanchang 330006, P. R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Cai
authors.affiliation: Department of Oncology, The Second Affiliated Hospital of Nanchang University Nanchang 330006, P. R. China.
authors.forename: An-Wen
authors.initials: AW
authors.lastname: Liu
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D003374:Coumarins / Q000494:pharmacology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D015394:Molecular Structure; D010051:Ovarian Neoplasms / Q000473:pathology*
chemical_list: C000603192:3-cinnamoyl-4-hydroxy-6-methyl-2H-pyran-2-one; D003374:Coumarins
keywords: Coumarins; annexin V-FITC; apoptosis; flow cytometry; mitochondrial membrane potential