Gelasia Tomentosa Butanolic Extract Exhibits Antiproliferative Activity Against MCF-7 Breast Cancer Cells: Mechanistic Insights into Synergistic EGFR and TOP2A Targeting

• Amjed Abbawe Salih

  College of Dentistry, Kirkuk University, Iraq

• Amenh Mohammed Abdulrahman

  Al-Iraqia University, College of Medicine, Department of Pharmacology, Baghdad, Iraq

• Ali Mala Khedir Galalaey

  College of Agricultural Engineering Sciences, University of Salahaddin-Erbil, Kurdistan Region, Iraq

• Waleed Khalid Ahmed

  Department of Chemistry and Biochemistry, College of Medicine, University of Fallujah, Iraq

• Shatha H. Shaker

  Department of Biology, College of Education for Pure Sciences, Tikrit University, Iraq

• Youssef Shakuri Yasin

  Bilad Alrafidain University, Iraq

• Ahmed Mohsin Abdulkhadim

  Kamal Al-Samarrai Hospital, Ministry of Health, Iraq

• Azal Hamoody Jumaa

  Iraqi National Cancer Research Center/University of Baghdad, Iraq

Objective: This preliminary study investigated the in vitro cytotoxicity and selectivity of the Gelasia tomentosa butanolic extract (Gt-BuOH) against MCF-7 breast cancer cells, using HFF fibroblasts as a non-malignant control, and explored associations between treatment and changes in p-EGFR, TOP2A, and TNF-α protein levels. Molecular docking was performed to generate hypotheses.

Materials and Methods: Gt-BuOH (13.6% w/w fatty extract) was prepared by liquid-liquid partitioning of 650 g of powdered aerial parts. Chemical composition was characterized by GC-MS with retention index matching; compounds with similarity index ≥70% were tentatively identified. Cytotoxicity was determined using the MTT assay for cell viability at 0.1–1000 µg/mL in the MCF-7 cancer cell line and healthy HFF cells after 24 h and 72 h of incubation. Quantification of p-EGFR, TOP2A, and TNF-α was performed by ELISA. Key phytoconstituents were docked against these targets, with binding affinities estimated using AutoDock Vina.

Results: Gt-BuOH inhibited MCF-7 cells in a concentration- and time-dependent manner (2.0 ± 0.2% to 75.0 ± 0.3% at 72 h; IC₅₀ = 431.8 µg/mL) with low toxicity towards HFF (IC₅₀ > 1000 µg/mL; selectivity index > 2.32). GC-MS identified methyl oleate (71.62% of total ion chromatogram area), propanoic anhydride, and methyl butyrate as the principal constituents; the profile is dominated by fatty-acid derivatives rather than phenolics. Gt-BuOH treatment was associated with reduced p-EGFR, TOP2A, and TNF-α levels in MCF-7 cells but not in HFF cells. Docking predicted weaker individual binding affinities for the three constituents than for reference inhibitors; no synergy or multi-target advantage is claimed.

Conclusion: These preliminary findings support further investigation of Gt-BuOH as a source of fatty-acid-derived antiproliferative agents, but bioassay-guided fractionation, additional cell lines, and functional validation are required before any therapeutic claims can be made.

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