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...

Full description

Saved in:
Bibliographic Details
Main Authors: Kuo-Ting Chang, Yu-Cheng Chen, Yi Lien, Yen-Hua Huang, Cheng-Yang Huang
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Plants
Subjects:
Online Access:https://www.mdpi.com/2223-7747/14/10/1426
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849327078013403136
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
collection DOAJ
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
publisher MDPI AG
record_format Article
series Plants
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
work_keys_str_mv AT kuotingchang inhibitionofrpa32andcytotoxiceffectsofthecarnivorousplantisarraceniapurpureairootextractinnonsmallcelllungcancercells
AT yuchengchen inhibitionofrpa32andcytotoxiceffectsofthecarnivorousplantisarraceniapurpureairootextractinnonsmallcelllungcancercells
AT yilien inhibitionofrpa32andcytotoxiceffectsofthecarnivorousplantisarraceniapurpureairootextractinnonsmallcelllungcancercells
AT yenhuahuang inhibitionofrpa32andcytotoxiceffectsofthecarnivorousplantisarraceniapurpureairootextractinnonsmallcelllungcancercells
AT chengyanghuang inhibitionofrpa32andcytotoxiceffectsofthecarnivorousplantisarraceniapurpureairootextractinnonsmallcelllungcancercells