地胆草和大果木蝴蝶叶提取物的组合通过下调雌激素受体α、核因子E2相关因子2和磷脂酰肌醇-3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路促进抗癌活性。

The combination of Elephantopus scaber and Phaleria macrocarpa leaves extract promotes anticancer activity via downregulation of ER-α, Nrf2 and PI3K/AKT/mTOR pathway.

作者信息

Christina Yuyun Ika, Rifa'i Muhaimin, Widodo Nashi, Djati Muhammad Sasmito

机构信息

Doctoral Program, Department of Biology, Faculty of Mathematics and Natural Sciences, Brawijaya University, Malang 65145, East Java, Indonesia.

Department of Biology, Faculty of Mathematics and Natural Sciences, Brawijaya University, Malang 65145, East Java, Indonesia.

出版信息

J Ayurveda Integr Med. 2022 Oct-Dec;13(4):100674. doi: 10.1016/j.jaim.2022.100674. Epub 2022 Dec 8.

DOI:10.1016/j.jaim.2022.100674

PMID:36502785

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9950941/

Abstract

BACKGROUND

Elephantopus scaber and Phaleria macrocarpa have recently been interested as novel anticancer agents. However, there was no scientific evaluation of the anticancer effect of both plant combinations.

OBJECTIVE

This study investigated the potential anticancer effects of combined E. scaber and P. macrocarpa leaves extract on human breast cancer cells lines.

MATERIALS AND METHODS

T47D cells were treated with the combination of E. scaber and each part of P. macrocarpa (leaves/EL, mesocarp/EM, seed/ES and pericarp/EP). T47D cells were then exposed to three ratios (1:1, 2:1, and 1:2) of the best combination for 24, 48, and 72 h. The cell viability of T47D and TIG-1 cells was assessed using WST-1 assay. The apoptotic hallmarks were determined using FITC Annexin V-PI staining and DNA fragmentation assay. The cell proliferation and cell cycle profiles were analyzed using CFSE (carboxyfluorescein succinimidyl ester) and Propidium iodide-flowcytometry assays. The relative number of p-ERα, p-Nrf2, p-PI3K, p-AKT, and p-mTOR were assessed using flow cytometry. The molecular docking analysis was also performed to confirm the mechanism of the extract in silico.

RESULTS

The combination of E. scaber and P. macrocarpa leaves (EL) possessed strong cytotoxic activity (p < 0.05) than other combination groups and cisplatin. EL showed selective killing only to T47D cells. EL at a ratio of 2:1 potentially suppressed the cell viability and cell division, induced apoptosis, and arrested the cell cycle of T47D cells by triple inhibiting the p-Nrf2, p-ERα, and p-PI3K/AKT/mTOR signaling pathway. Molecular docking analysis confirmed that the possible mechanism of EL to reduce T47D cell growth was by inhibiting ERα and Nrf2-complex, resulting in the reduction in the crosstalk effect of Nrf2, ERα and PI3K/AKT/mTOR pathways.

CONCLUSION

The combination of leaf extracts from E. scaber and P. macrocarpa caused cell death in breast cancer cells T47D and not in normal cells TIG-1; hence has the potential to show anticancer efficacy in preclinical and clinical trials.

摘要

背景

最近，地胆草和大果木姜子作为新型抗癌药物受到关注。然而，对于这两种植物组合的抗癌效果尚无科学评估。

目的

本研究调查了地胆草和大果木姜子叶提取物联合使用对人乳腺癌细胞系的潜在抗癌作用。

材料与方法

用地胆草与大果木姜子各部分（叶/EL、中果皮/EM、种子/ES 和果皮/EP）的组合处理 T47D 细胞。然后将 T47D 细胞暴露于最佳组合的三种比例（1:1、2:1 和 1:2）下 24、48 和 72 小时。使用 WST-1 试验评估 T47D 和 TIG-1 细胞的细胞活力。使用 FITC 膜联蛋白 V-碘化丙啶染色和 DNA 片段化试验确定凋亡特征。使用羧基荧光素琥珀酰亚胺酯（CFSE）和碘化丙啶流式细胞术试验分析细胞增殖和细胞周期谱。使用流式细胞术评估 p-ERα、p-Nrf2、p-PI3K、p-AKT 和 p-mTOR 的相对数量。还进行了分子对接分析以在计算机上确认提取物的作用机制。

结果

地胆草和大果木姜子叶（EL）的组合比其他组合组和顺铂具有更强的细胞毒性活性（p < 0.05）。EL 仅对 T47D 细胞表现出选择性杀伤作用。2:1 比例的 EL 可能通过三重抑制 p-Nrf2、p-ERα 和 p-PI3K/AKT/mTOR 信号通路来抑制 T47D 细胞的细胞活力和细胞分裂，诱导凋亡并使细胞周期停滞。分子对接分析证实，EL 降低 T47D 细胞生长的可能机制是通过抑制 ERα 和 Nrf2 复合物，从而减少 Nrf2、ERα 和 PI3K/AKT/mTOR 通路的相互作用效应。