7-Hydroxydehydronuciferine induces human melanoma death via triggering autophagy and apoptosis

• Published in: Experimental dermatology 2015-12, Vol.24 (12), p.930-935
• Main Authors: Wu, Pei-Fang, Chiu, Chien-Chih, Chen, Chung-Yi, Wang, Hui-Min David
• Format: Article
• Language: English
• Subjects: 7-hydroxydehydronuciferine; Animals; Annexin V; Antineoplastic Agents, Phytogenic - chemistry; Antineoplastic Agents, Phytogenic - pharmacology; Apoptosis; Apoptosis - drug effects; Aporphines - chemistry; Aporphines - pharmacology; Autophagy; Autophagy - drug effects; Cancer; Cell cycle; Cell death; Cell Line, Tumor; Cell membranes; Cell proliferation; Cell Proliferation - drug effects; Drugs, Chinese Herbal - chemistry; Drugs, Chinese Herbal - pharmacology; Female; Fibroblasts; Flow cytometry; Fluorescein isothiocyanate; G2 Phase Cell Cycle Checkpoints - drug effects; Humans; Keratinocytes; Keratinocytes - cytology; Keratinocytes - drug effects; Medicine, Chinese Traditional; Melanocytes; Melanoma; Melanoma - drug therapy; Melanoma - metabolism; Melanoma - pathology; metastasis; Mice; Mice, Inbred BALB C; Mice, Nude; Microtubule-Associated Proteins - metabolism; Mitochondrial Proteins - metabolism; Nelumbo - chemistry; Organelles; Plants, Medicinal - chemistry; Propidium iodide; Tumors; Vacuoles; Wound healing; Xenograft Model Antitumor Assays; Xenografts
• ISSN: 0906-6705; 1600-0625
• DOI: 10.1111/exd.12805

Melanoma is the deadliest cancer. We identified 7‐hydroxydehydronuciferine (7‐HDNF) isolated from the leaves of Nelumbo nucifera Gaertn cv. Rosa‐plena to be a bio‐active agent that antagonizes melanoma tumor growth in mice xenograft model in vivo. Cell proliferation assay demonstrated strong anticancer effects of 7‐HDNF to exhibit a dose‐dependent behaviour and displayed minor cytotoxicities on normal human skin cells, including epidermal keratinocytes and melanocytes, and dermal fibroblasts. With acridine orange (AO) staining and flow analysis, we found 7‐HDNF induced the formation of intracellular vacuoles and the augmentation of acidic vesicular organelles (AVO). The apoptotic cell death ratio was measured via two‐dimensional flow cytometry by annexin V‐fluorescein isothiocyanate (FITC)/propidium iodide (PI) double stained to confirm the cellular membrane asymmetry lost. One‐dimensional flow cytometric analysis showed 7‐HDNF increased the cellular arrest in cell cycle at the G2/M phase. Through Western blot examinations, protein expressions were discovered to verify autophagy and apoptosis response mechanisms sharing the associated pathways. Finally, 7‐HDNF presented a high‐quality antimigratory activity in wound‐healing assay. Overall, 7‐HDNF presented high‐quality anticancer bio‐functions and inhibited melanoma tumor growth in vivo and in vitro.