Chang Kuo-Ting, Chen Yu-Cheng, Lien Yi, Huang Yen-Hua, Huang Cheng-Yang
Division of Translational Medicine, Department of Research and Development, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan 330, Taiwan.
Division of Pulmonary Medicine, Department of Internal Medicine, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan 330, Taiwan.
Plants (Basel). 2025 May 9;14(10):1426. doi: 10.3390/plants14101426.
The carnivorous plant 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 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 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 of 13.6 μg/mL. AlphaFold structural prediction and molecular docking studies indicated that major bioactive compounds in , 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 root extract against NSCLC is highlighted, supporting further investigation into its therapeutic applications.
这种食虫植物传统上已被用于各种民族植物学应用，包括治疗2型糖尿病和类似肺结核的症状。本研究调查了根提取物（Sp-R）对人非小细胞肺癌（NSCLC）细胞系（包括H1975、H838和A549）的细胞毒性作用，重点关注其对细胞存活、凋亡、增殖和迁移的影响。此外，还评估了其抑制人RPA32（huRPA32，DNA复制中的关键蛋白）单链DNA结合活性的能力。使用各种溶剂（水、甲醇、乙醇和丙酮）制备了来自不同植物部位（叶、茎和根）的提取物，并使用染色质凝聚试验筛选其诱导凋亡的潜力。其中，丙酮提取的根部分（Sp-R-A）表现出最有效的促凋亡作用。MTT试验证明对NSCLC细胞有剂量依赖性细胞毒性作用，H1975的IC值为33.74μg/mL，H838为60.79μg/mL，A549为66.52μg/mL。迁移和克隆形成试验进一步表明，Sp-R-A以剂量依赖性方式显著抑制癌细胞迁移和集落形成。此外，Sp-R-A与表皮生长因子受体（EGFR）抑制剂阿法替尼联合使用时可增强凋亡，提示可能存在协同效应。电泳迁移率变动分析证实，Sp-R-A显著抑制huRPA32的DNA结合活性，IC值为13.6μg/mL。AlphaFold结构预测和分子对接研究表明，该植物中的主要生物活性化合物，包括α-香树脂醇、熊果酸和桦木醛，与huRPA32的DNA结合结构域强烈相互作用，可能有助于其抑制作用。总体而言，这些发现表明huRPA32是Sp-R-A的潜在分子靶点，突出了该植物根提取物对NSCLC的抗癌潜力，支持对其治疗应用进行进一步研究。
