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. 2025 Jan 25;17(1):e77985.
doi: 10.7759/cureus.77985. eCollection 2025 Jan.

The Anti-Cancer Effects of Arborinine from Ruta graveolens L. on Michigan Cancer Foundation-7 (MCF-7) Breast Cancer Cells: Inhibition of Cell Growth and Induction of Apoptosis

Vahid Zangouri  1 , Hamid Zaferani Arani  2 , Seyed Alireza Salimi Tabatabaee  3
Affiliations

The Anti-Cancer Effects of Arborinine from Ruta graveolens L. on Michigan Cancer Foundation-7 (MCF-7) Breast Cancer Cells: Inhibition of Cell Growth and Induction of Apoptosis

Vahid Zangouri et al. Cureus. .

Abstract

Background: Arborinine, a plant-derived alkaloid, has shown potential cytotoxic effects against various cancer cell lines. This study aims to evaluate the cytotoxicity and apoptosis effects of arborinine on breast cancer (Michigan Cancer Foundation-7 (MCF-7)) and human embryonic kidney (HEK293) as normal cell lines.

Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to assess the inhibitory concentration of 50% (IC50) after 24 and 48 hours of treatment of HEK293 and MCF-7 cell lines with arborinine. Apoptosis was evaluated through Annexin V/PI staining, and gene expressions including BAX, BCL-2, P53, PARP, and caspases (i.e., 3, 8, and 9) were analyzed via quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, intracellular reactive oxygen species (ROS) levels were measured using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence.

Results: The MTT assay results indicated a dose-dependent reduction in cell viability for both HEK293 and MCF-7 cells following treatment with arborinine. The viability of HEK293 cells decreased significantly (P=0.038) at concentrations above 150 µg/mL, while IC50 for MCF-7 cells was 50 µg/mL and 25 µg/mL for 24 and 48 hours, respectively. Annexin V staining revealed apoptosis rates of 9.36% in untreated MCF-7 cells, increasing to 52.3% post-treatment. Arborinine treatment upregulated pro-apoptotic factors, including BAX, PARP, and P53, while downregulating BCL-2. Additionally, arborinine increased ROS levels by approximately 1.3-fold and decreased glutathione (GSH) levels, while enhancing superoxide dismutase (SOD) activity.

Conclusion: This study shows that arborinine reduces cell viability and induces apoptosis in MCF-7 breast cancer cells by modulating key apoptotic pathways. Its effectiveness at lower concentrations in cancerous cells highlights its potential as a promising therapeutic agent in oncology.

Keywords: anti-tumor; apoptosis; arborinine; cytotoxic effect; mcf-7; ruta graveolens l.

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Conflict of interest statement

Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. Ethics Committee of Shiraz University of Medical Sciences issued approval IR.SUMS.MED.REC.1403.291. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work.

Figures

Figure 1
Figure 1. Effect of arborinine on cell viability in HEK293 and MCF-7 cells.
The percentage of viable HEK293 cells after 24 (A) and 48 hours (B) of treatment with various concentrations of arborinine indicates that the IC50 of arborinine was 150 µg/mL. However, the IC50 of arborinine for MCF-7 cells after 24 (C) and 48 hours (D) were 50 and 25 µg/mL, respectively. Statistical analysis indicates significant differences (*P<0.05) compared to untreated cells.
Figure 2
Figure 2. Annexin V staining in MCF-7 cells with no treatment (A) and treated with arborinine (B).
PI, propidium iodide; FITC, fluorescein isothiocyanate
Figure 3
Figure 3. The relative expression ratio of apoptosis-related factors (BAX, BCL-2, PARP, P53, caspase 3, caspase 8, and caspase 9) in MCF-7 cells after 48 hours, induced by arborinine at a concentration of 25 µg/mL.
*P<0.05 compared to control.
Figure 4
Figure 4. Relative expression ratios of oxidative and anti-oxidative factors in MCF-7 cells after 48 hours of treatment with arborinine at 25 µg/mL. The levels of SOD, GSH, and ROS were measured.
SOD, superoxide dismutase; GSH, glutathione; ROS, reactive oxygen species *P<0.05 denotes statistical significance when compared to the control group.
See this image and copyright information in PMC

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