AI Article Synopsis
- The study investigates how Houttuynia cordata Thunb (HCT) extract causes cell death in human melanoma A375 cells, focusing on apoptosis mechanisms.
- Key experimental methods included the MTT assay for cell viability, Hoechst staining for apoptosis confirmation, and various assays to measure mRNA and protein levels related to apoptotic pathways.
- Results showed HCT activates caspase-3 and caspase-8, leading to increased cell death, and involves p38 MAPK phosphorylation, indicating a caspase-dependent apoptosis pathway linked to reduced HMGB1 release.
Article Abstract
Background/aim: To analyze the apoptotic effect of Houttuynia cordata Thunb (HCT) extract on human melanoma A375 cells and its underlying mechanisms.
Materials And Methods: The effects of HCT on cell death were determined using the MTT assay. Hoechst 33342 staining was conducted to confirm the detection of cell apoptosis. Caspase-3 and caspase-8 mRNA and cleaved protein levels were investigated by RT-PCR and western blotting, respectively. The release of high mobility group box 1 (HMGB1) and phosphorylation of mitogen-activated protein kinase (MAPK) were determined by ELISA.
Results: Caspase-3 and caspase-8 specific inhibitors suppressed HCT-induced cell death. HCT increased caspase-3 and caspase-8 mRNA, protein levels, and caspase activities in a concentration- and time-dependent manner. HCT induced MAPK phosphorylation in a time-dependent fashion. Pretreatment of cells with a selective inhibitor of p38 MAPK reduced apoptosis and reversed the levels of HMGB1 release in response to HCT treatment.
Conclusion: HCT induces A375 programmed cell death by activating the caspase-dependent pathway and by p38 phosphorylation associated with HMGB1 reduction.
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| http://dx.doi.org/10.21873/anticanres.12119 | DOI Listing |
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