Insight into OroxylinA-7- O -β-d-Glucuronide-Enriched Oroxylum indicum Bark Extract in Oral Cancer HSC-3 Cell Apoptotic Mechanism: Role of Mitochondrial Microenvironment

• Published in: Molecules (Basel, Switzerland) Switzerland), 2021-12, Vol.26 (24), p.7430
• Main Authors: Kameyanda Poonacha, Sharmila, Harishkumar, Madhyastha, Radha, Madhyastha, Varadarajan, Remya, Nalilu, Suchetha Kumari, Shetty, Shilpa Sharathraj, Shetty, Praveen Kumar, Chandrashekharappa, Revanasiddappa Bistuvalli, Sreenivas, Mahendra Gowdru, Bhandary Bavabeedu, Satheesh Kumar
• Format: Article
• Language: English
• Subjects: Antiinfectives and antibacterials; Antineoplastic Agents, Phytogenic - chemistry; Antineoplastic Agents, Phytogenic - isolation & purification; Antineoplastic Agents, Phytogenic - pharmacology; Apoptosis; Apoptosis - drug effects; Assaying; Bark; Bignoniaceae - chemistry; Binding sites; Biodistribution; Bronchitis; Calcein; Cancer therapies; Caspase-3; Cell cycle; Cell death; Cell Line, Tumor; Cell number; Cell proliferation; Cell Proliferation - drug effects; Cell viability; Cytochemistry; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA damage; Dose-Response Relationship, Drug; Drug dosages; Drug Screening Assays, Antitumor; Elongation; Flavonoids; Fluorescence; HSC-3 cell; Humans; Immunoblotting; Inflammation; Microenvironments; Microorganisms; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; mitochondrial membrane potential; mitochondrial stress; Molecular docking; Molecular Structure; Morphology; Necrosis; Oral cancer; Oroxylum indicum; Permeability; Plant Bark - chemistry; Plant extracts; Plant Extracts - chemistry; Plant Extracts - isolation & purification; Plant Extracts - pharmacology; Skin cancer; Staining; Structure-Activity Relationship; Toxicity; Tumor Microenvironment - drug effects
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules26247430

, of the Bignoniaceae family, has various ethnomedical uses such as an astringent, anti-inflammatory, anti-bronchitis, anti-helminthic and anti-microbial, including anticancer properties. The druggability of OI stem bark extract was determined by its molecular docking interactions with PARP and Caspase-3, two proteins involved in cell survival and death. Note that 50 µg/mL of extract (OIE) showed a significant ( < 0.05%) toxicity to HSC-3 cells. MTT aided cell viability and proliferation assay demonstrated that 50 µg/mL of OIE displayed significant ( < 0.5%) reduction in cell number at 4 h of incubation time. Cell elongation and spindle formation was noticed when HSC-3 cells were treated with 50 µg/mL of OIE. OIE initiated DNA breakage and apoptosis in HSC-3 cells, as evident from DNA ladder assay and calcein/EB staining. Apoptosis potential of OIE is confirmed by flow cytometer and triple-staining (live cell/apoptosis/necrosis) assay. Caspase-3/7 fluorescence quenching (LANCE) assay demonstrated that 50 µg/mL of OIE significantly enhanced the RFU of caspases-3/7, indicating that the apoptosis potential of OIE is probably through the activation of caspases. Immuno-cytochemistry of HSC-3 cells treated with 50 µg/mL of OIE showed a significant reduction in mitochondrial bodies as well as a reduction in RFU in 60 min of incubation time. Immunoblotting studies clearly showed that treatment of HSC-3 cells with OI extract caused caspase-3 activation and PARP deactivation, resulting in apoptotic cell death. Overall, our data indicate that OIE is an effective apoptotic agent for human squamous carcinoma cells and it could be a future cancer chemotherapeutic target.