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

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.

Angiogenesis; Apoptosis; Breast Cancer; Cytotoxicity; MDA-MB-231; Rocaglamide; Triple-negative breast cancer.