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通过细胞培养和分子方法评估罗克拉酰胺对MDA-MB-231人乳腺腺癌细胞的影响。

Evaluation of the impact of Roclaglamide on MDA-MB-231 human breast adenocarcinoma cells by cell culture and molecular approaches.

作者信息

Sandakli Ruaa, Saadoun Ismail, Al-Joubori Ban, Khattak Muhammad Nasir Khan

机构信息

Department of Applied Biology, College of Sciences, University of Sharjah, P.O. Box 27272, Sharjah, UAE.

Senior Academic Researcher, Institute of Community Medicine, SHIP/KEF, Greifswald, Germany.

出版信息

Saudi Pharm J. 2025 Jul 8;33(4):22. doi: 10.1007/s44446-025-00023-5.

Abstract

1H - 2,3,3a,8b-tetrahydrocyclopenta[b]benzofuran, known as Rocaglamide (RocA) has gained recognition for its potential as a plant-derived therapeutic agent in cancer treatment. This study evaluates the impact of RocA on MDA-MB-231 human breast adenocarcinoma cells. Rocaglamide's effect was comprehensively evaluated through cell culture and molecular approaches with a series of experimental procedures. Cell viability was assessed using the MTT assay which determined the compound's cytotoxic effects. Following this, apoptosis induction in cancer cells was examined through Annexin V-FITC staining and flow cytometric analysis, providing insight into Rocaglamide's apoptotic mechanism of action. Additionally, the proliferation of cancer cells was evaluated by Propidium Iodide (PI) staining, elucidating Rocaglamide's antiproliferative effects on breast cancer cells. Furthermore, protein expression levels were analyzed using the Human XL Oncology Array Kit, shedding light on Rocaglamide's molecular targets and effect on various signaling pathways as demonstrated by how rocaglamide particularly attenuate the activity of proteins crucial for angiogenesis and tumor progression. MTT assay revealed a time-dependent decrease in cell viability. When treating MDA-MB-231 breast cancer cells with increasing concentrations of RocA, significant morphological changes were observed. Wound healing assay to evaluate the impact of RocA on cell migration indicated the induction of slight apoptosis in breast cancer cells. Flow cytometry-based apoptosis assay revealed that the total cell death in treated cells reached around 15%, compared to 7.9% in the untreated group. The expression of key oncogenic proteins after treatment of MDA-MB-231 cells with RocA indicated a noticeable reduction of VEGFA, AXL, and PAI-1 when compared to the untreated control. However, SNAIL1 and Endoglin increased significantly relative to untreated controls. Collectively, the findings demonstrate Rocaglamide's potential as a therapeutic agent for breast cancer and offer valuable insights into its mechanisms of action.

摘要

1H - 2,3,3a,8b - 四氢环戊并[b]苯并呋喃,即罗卡酰胺(RocA),因其作为一种植物源治疗剂在癌症治疗中的潜力而受到认可。本研究评估了罗卡酰胺对MDA - MB - 231人乳腺腺癌细胞的影响。通过细胞培养和分子方法以及一系列实验程序,对罗卡酰胺的作用进行了全面评估。使用MTT法评估细胞活力,该方法确定了该化合物的细胞毒性作用。在此之后,通过膜联蛋白V - FITC染色和流式细胞术分析检测癌细胞中的凋亡诱导情况,从而深入了解罗卡酰胺的凋亡作用机制。此外,通过碘化丙啶(PI)染色评估癌细胞的增殖情况,阐明罗卡酰胺对乳腺癌细胞的抗增殖作用。此外,使用人类XL肿瘤学阵列试剂盒分析蛋白质表达水平,揭示罗卡酰胺的分子靶点以及对各种信号通路的影响,如罗卡酰胺特别减弱了对血管生成和肿瘤进展至关重要的蛋白质的活性。MTT法显示细胞活力呈时间依赖性下降。当用浓度不断增加的罗卡酰胺处理MDA - MB - 231乳腺癌细胞时,观察到明显的形态变化。用于评估罗卡酰胺对细胞迁移影响的伤口愈合试验表明乳腺癌细胞中诱导了轻微凋亡。基于流式细胞术的凋亡试验显示,处理组细胞的总细胞死亡率达到约15%,而未处理组为7.9%。用罗卡酰胺处理MDA - MB - 231细胞后,关键致癌蛋白的表达表明,与未处理对照相比,VEGFA、AXL和PAI - 1明显减少。然而,与未处理对照相比,SNAIL1和内皮糖蛋白显著增加。总的来说,这些发现证明了罗卡酰胺作为乳腺癌治疗剂的潜力,并为其作用机制提供了有价值的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f30c/12238422/6484f340a16e/44446_2025_23_Fig1_HTML.jpg

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