Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase Proteins
The carnivorous pitcher plants of the genus <i>Nepenthes</i> have long been known for their ethnobotanical applications. In this study, we prepared various extracts from the pitcher, stem, and leaf of <i>Nepenthes miranda</i> using 100% ethanol and assessed their inhibitory e...
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MDPI AG
2024-03-01
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| author | Ching-Yi Lee Yu-Cheng Chen Yen-Hua Huang Yi Lien Cheng-Yang Huang |
| author_facet | Ching-Yi Lee Yu-Cheng Chen Yen-Hua Huang Yi Lien Cheng-Yang Huang |
| author_sort | Ching-Yi Lee |
| collection | DOAJ |
| description | The carnivorous pitcher plants of the genus <i>Nepenthes</i> have long been known for their ethnobotanical applications. In this study, we prepared various extracts from the pitcher, stem, and leaf of <i>Nepenthes miranda</i> using 100% ethanol and assessed their inhibitory effects on key enzymes related to skin aging, including elastase, tyrosinase, and hyaluronidase. The cytotoxicity of the stem extract of <i>N. miranda</i> on H838 human lung carcinoma cells were also characterized by effects on cell survival, migration, proliferation, apoptosis induction, and DNA damage. The cytotoxic efficacy of the extract was enhanced when combined with the chemotherapeutic agent 5-fluorouracil (5-FU), indicating a synergistic effect. Flow cytometry analysis suggested that the stem extract might suppress H838 cell proliferation by inducing G2 cell cycle arrest, thereby inhibiting carcinoma cell proliferation. Gas chromatography–mass spectrometry (GC–MS) enabled the tentative identification of the 15 most abundant compounds in the stem extract of <i>N. miranda</i>. Notably, the extract showed a potent inhibition of the human RPA32 protein (huRPA32), critical for DNA replication, suggesting a novel mechanism for its anticancer action. Molecular docking studies further substantiated the interaction between the extract and huRPA32, highlighting bioactive compounds, especially the two most abundant constituents, stigmast-5-en-3-ol and plumbagin, as potential inhibitors of huRPA32’s DNA-binding activity, offering promising avenues for cancer therapy. Overall, our findings position the stem extract of <i>N. miranda</i> as a promising source of natural compounds for anticancer therapeutics and anti-skin-aging treatments, warranting further investigation into its molecular mechanisms and potential clinical applications. |
| first_indexed | 2024-04-24T17:53:37Z |
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| last_indexed | 2024-04-24T17:53:37Z |
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| spelling | doaj.art-394d78291c664fb2987f2aee3b61b02a2024-03-27T14:00:40ZengMDPI AGPlants2223-77472024-03-0113679710.3390/plants13060797Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase ProteinsChing-Yi Lee0Yu-Cheng Chen1Yen-Hua Huang2Yi Lien3Cheng-Yang Huang4Department of Internal Medicine, Tao Yuan General Hospital, Ministry of Health and Welfare, Taoyuan 330, TaiwanDepartment of Internal Medicine, Tao Yuan General Hospital, Ministry of Health and Welfare, Taoyuan 330, TaiwanDepartment of Biomedical Sciences, Chung Shan Medical University, Taichung City 402, TaiwanDepartment of Biological Sciences, Purdue University, West Lafayette, IN 47907, USADepartment of Biomedical Sciences, Chung Shan Medical University, Taichung City 402, TaiwanThe carnivorous pitcher plants of the genus <i>Nepenthes</i> have long been known for their ethnobotanical applications. In this study, we prepared various extracts from the pitcher, stem, and leaf of <i>Nepenthes miranda</i> using 100% ethanol and assessed their inhibitory effects on key enzymes related to skin aging, including elastase, tyrosinase, and hyaluronidase. The cytotoxicity of the stem extract of <i>N. miranda</i> on H838 human lung carcinoma cells were also characterized by effects on cell survival, migration, proliferation, apoptosis induction, and DNA damage. The cytotoxic efficacy of the extract was enhanced when combined with the chemotherapeutic agent 5-fluorouracil (5-FU), indicating a synergistic effect. Flow cytometry analysis suggested that the stem extract might suppress H838 cell proliferation by inducing G2 cell cycle arrest, thereby inhibiting carcinoma cell proliferation. Gas chromatography–mass spectrometry (GC–MS) enabled the tentative identification of the 15 most abundant compounds in the stem extract of <i>N. miranda</i>. Notably, the extract showed a potent inhibition of the human RPA32 protein (huRPA32), critical for DNA replication, suggesting a novel mechanism for its anticancer action. Molecular docking studies further substantiated the interaction between the extract and huRPA32, highlighting bioactive compounds, especially the two most abundant constituents, stigmast-5-en-3-ol and plumbagin, as potential inhibitors of huRPA32’s DNA-binding activity, offering promising avenues for cancer therapy. Overall, our findings position the stem extract of <i>N. miranda</i> as a promising source of natural compounds for anticancer therapeutics and anti-skin-aging treatments, warranting further investigation into its molecular mechanisms and potential clinical applications.https://www.mdpi.com/2223-7747/13/6/797<i>Nepenthes</i>anticancerH838 lung carcinomaanti-skin agingelastaseRPA |
| spellingShingle | Ching-Yi Lee Yu-Cheng Chen Yen-Hua Huang Yi Lien Cheng-Yang Huang Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase Proteins Plants <i>Nepenthes</i> anticancer H838 lung carcinoma anti-skin aging elastase RPA |
| title | Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase Proteins |
| title_full | Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase Proteins |
| title_fullStr | Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase Proteins |
| title_full_unstemmed | Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase Proteins |
| title_short | Cytotoxicity and Multi-Enzyme Inhibition of <i>Nepenthes miranda</i> Stem Extract on H838 Human Non-Small Cell Lung Cancer Cells and RPA32, Elastase, Tyrosinase, and Hyaluronidase Proteins |
| title_sort | cytotoxicity and multi enzyme inhibition of i nepenthes miranda i stem extract on h838 human non small cell lung cancer cells and rpa32 elastase tyrosinase and hyaluronidase proteins |
| topic | <i>Nepenthes</i> anticancer H838 lung carcinoma anti-skin aging elastase RPA |
| url | https://www.mdpi.com/2223-7747/13/6/797 |
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