Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer Cells
The carnivorous plant <i>Sarracenia purpurea</i> has been traditionally used in various ethnobotanical applications, including treatments for type 2 diabetes and tuberculosis-like symptoms. This study investigates the cytotoxic effects of <i>S. purpurea</i> root extract (Sp-R...
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2025-05-01
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| author | Kuo-Ting Chang Yu-Cheng Chen Yi Lien Yen-Hua Huang Cheng-Yang Huang |
| author_facet | Kuo-Ting Chang Yu-Cheng Chen Yi Lien Yen-Hua Huang Cheng-Yang Huang |
| author_sort | Kuo-Ting Chang |
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| description | The carnivorous plant <i>Sarracenia purpurea</i> has been traditionally used in various ethnobotanical applications, including treatments for type 2 diabetes and tuberculosis-like symptoms. This study investigates the cytotoxic effects of <i>S. purpurea</i> root extract (Sp-R) on human non-small-cell lung cancer (NSCLC) cell lines, including H1975, H838, and A549, focusing on its impact on cell survival, apoptosis, proliferation, and migration. Additionally, its ability to inhibit the single-stranded DNA-binding activity of human RPA32 (huRPA32), a key protein in DNA replication, was evaluated. Extracts from different plant parts (leaf, stem, and root) were prepared using various solvents (water, methanol, ethanol, and acetone) and screened for apoptosis-inducing potential using the chromatin condensation assay. Among these, the acetone-extracted root fraction (Sp-R-A) exhibited the most potent pro-apoptotic effects. The MTT assay demonstrated a dose-dependent cytotoxic effect on NSCLC cells, with IC<sub>50</sub> values of 33.74 μg/mL for H1975, 60.79 μg/mL for H838, and 66.52 μg/mL for A549. Migration and clonogenic assays further revealed that Sp-R-A significantly inhibited cancer cell migration and colony formation in a dose-dependent manner. Moreover, Sp-R-A enhanced apoptosis when combined with the EGFR inhibitor afatinib, suggesting a potential synergistic effect. The electrophoretic mobility shift assay confirmed that Sp-R-A significantly inhibited the DNA-binding activity of huRPA32, with an IC<sub>50</sub> of 13.6 μg/mL. AlphaFold structural prediction and molecular docking studies indicated that major bioactive compounds in <i>S. purpurea</i>, including α-amyrin, ursolic acid, and betulinaldehyde, strongly interact with the DNA-binding domain of huRPA32, potentially contributing to its inhibitory effect. Overall, these findings suggest that huRPA32 is a potential molecular target of Sp-R-A and the anticancer potential of <i>S. purpurea</i> root extract against NSCLC is highlighted, supporting further investigation into its therapeutic applications. |
| format | Article |
| id | doaj-art-53a8bdc9dcaa4d36a48b5a4a69d22add |
| institution | Kabale University |
| issn | 2223-7747 |
| language | English |
| publishDate | 2025-05-01 |
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| spelling | doaj-art-53a8bdc9dcaa4d36a48b5a4a69d22add2025-08-20T03:47:58ZengMDPI AGPlants2223-77472025-05-011410142610.3390/plants14101426Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer CellsKuo-Ting Chang0Yu-Cheng Chen1Yi Lien2Yen-Hua Huang3Cheng-Yang Huang4Division of Translational Medicine, Department of Research and Development, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan 330, TaiwanDivision of Pulmonary Medicine, Department of Internal Medicine, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan 330, TaiwanDepartment of Biological Sciences, Purdue University, West Lafayette, IN 47907, USADepartment of Biomedical Sciences, Chung Shan Medical University, Taichung 402, TaiwanDepartment of Biomedical Sciences, Chung Shan Medical University, Taichung 402, TaiwanThe carnivorous plant <i>Sarracenia purpurea</i> has been traditionally used in various ethnobotanical applications, including treatments for type 2 diabetes and tuberculosis-like symptoms. This study investigates the cytotoxic effects of <i>S. purpurea</i> root extract (Sp-R) on human non-small-cell lung cancer (NSCLC) cell lines, including H1975, H838, and A549, focusing on its impact on cell survival, apoptosis, proliferation, and migration. Additionally, its ability to inhibit the single-stranded DNA-binding activity of human RPA32 (huRPA32), a key protein in DNA replication, was evaluated. Extracts from different plant parts (leaf, stem, and root) were prepared using various solvents (water, methanol, ethanol, and acetone) and screened for apoptosis-inducing potential using the chromatin condensation assay. Among these, the acetone-extracted root fraction (Sp-R-A) exhibited the most potent pro-apoptotic effects. The MTT assay demonstrated a dose-dependent cytotoxic effect on NSCLC cells, with IC<sub>50</sub> values of 33.74 μg/mL for H1975, 60.79 μg/mL for H838, and 66.52 μg/mL for A549. Migration and clonogenic assays further revealed that Sp-R-A significantly inhibited cancer cell migration and colony formation in a dose-dependent manner. Moreover, Sp-R-A enhanced apoptosis when combined with the EGFR inhibitor afatinib, suggesting a potential synergistic effect. The electrophoretic mobility shift assay confirmed that Sp-R-A significantly inhibited the DNA-binding activity of huRPA32, with an IC<sub>50</sub> of 13.6 μg/mL. AlphaFold structural prediction and molecular docking studies indicated that major bioactive compounds in <i>S. purpurea</i>, including α-amyrin, ursolic acid, and betulinaldehyde, strongly interact with the DNA-binding domain of huRPA32, potentially contributing to its inhibitory effect. Overall, these findings suggest that huRPA32 is a potential molecular target of Sp-R-A and the anticancer potential of <i>S. purpurea</i> root extract against NSCLC is highlighted, supporting further investigation into its therapeutic applications.https://www.mdpi.com/2223-7747/14/10/1426<i>Sarracenia purpurea</i>anticancerRPAssDNA bindingAlphaFoldNSCLC |
| spellingShingle | Kuo-Ting Chang Yu-Cheng Chen Yi Lien Yen-Hua Huang Cheng-Yang Huang Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer Cells Plants <i>Sarracenia purpurea</i> anticancer RPA ssDNA binding AlphaFold NSCLC |
| title | Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer Cells |
| title_full | Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer Cells |
| title_fullStr | Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer Cells |
| title_full_unstemmed | Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer Cells |
| title_short | Inhibition of RPA32 and Cytotoxic Effects of the Carnivorous Plant <i>Sarracenia purpurea</i> Root Extract in Non-Small-Cell Lung Cancer Cells |
| title_sort | inhibition of rpa32 and cytotoxic effects of the carnivorous plant i sarracenia purpurea i root extract in non small cell lung cancer cells |
| topic | <i>Sarracenia purpurea</i> anticancer RPA ssDNA binding AlphaFold NSCLC |
| url | https://www.mdpi.com/2223-7747/14/10/1426 |
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