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Thymoquinone-treated mouse mesenchymal stem cells-derived conditioned medium inhibits human breast cancer cells in
Pouria Hajmomeni1, Sajjad Sisakhtnezhad1, Ali Bidmeshkipour1
1Department of Biology, Faculty of Science, Razi University, Kermanshah, Iran.
Abstract:
Breast cancer is now the most prevalent cancer in females, therefore, it is essential to identify factors affecting its initiation and progression. Mesenchymal stem cells (MSCs) have received considerable attention in stem cell-based therapies and drug delivery applications. Because the therapeutic potential of MSCs is primarily achieved by their paracrine effects, thus identifying and employing bioactive molecules that promote the paracrine activity of MSCs is crucial for their efficient use in cancer treatment. Thymoquinone (TQ) has many biomedical properties, including anti-inflammatory, anti-diabetic, anti-aging, anti-cancer, etc. In addition, it has been found that TQ affects the self-renewal and immunomodulatory properties of MSCs. The present study aimed to investigate the effect of TQ-treated mouse bone marrow-derived MSCs conditioned medium (TQ-MSC-CM) on the biological characteristics of breast cancer cell line MCF7. MSCs were cultured and treated with TQ for 24 h. The TQ-MSC-CM and MSC-CM were collected, and their effects were investigated on ROS production, mitochondrial membrane potential (MMP), cell death, cell cycle, and migration of MCF7 cells by DCFDA-cellular ROS assay, Rhodamine-123 MMP assay, Annexin-PI staining and Caspase-3/7 activity assays, PI-staining and flow-cytometry, and in vitro wound healing assay, respectively. Moreover, the effects of TQ-MSC-CM and MSC-CM were studied on Cdk4, Sox2, c-Met, and Bcl2 gene expression by real-time PCR. Results demonstrated that MSC-CM and TQ-MSC-CM did not have a significant effect on the apoptosis induction in MCF7 cells; however, they significantly stimulated necrosis in the cells. Although TQ-MSC-CM promoted ROS production in MCF7 cells, it decreased the MMP of the cells. TQ-MSC-CM also induced Bcl2 anti-apoptosis gene expression and Casp-3/7 activity in cells. In addition, although MSC-CM induced MCF7 cells to enter the cell cycle, TQ-MSC-CM inhibited its progression. TQ-MSC-CM also downregulated the Cdk4 and Sox2 gene expression. Furthermore, TQ-MSC-CM induced the migration potential of MCF7 in a c-Met-independent manner. Altogether, we conclude that TQ may induce programmed necrosis and inhibits the proliferation and migration of the breast cancer cells by affecting the paracrine activity of MSCs.
Insights
Thymoquinone-treated mesenchymal stem cells (MSCs) conditioned medium induced necrosis and inhibited breast cancer cell proliferation and migration. This suggests TQ-MSC-CM is a promising approach for breast cancer therapy by modulating MSC paracrine activity.
Area of Science:
- Oncology
- Stem Cell Biology
- Biochemistry
Background:
- Breast cancer is the most common cancer in women, necessitating research into its initiation and progression factors.
- Mesenchymal stem cells (MSCs) are crucial for therapies due to their paracrine effects, requiring identification of molecules that enhance these effects.
- Thymoquinone (TQ) possesses various biomedical properties and influences MSC self-renewal and immunomodulatory functions.
Purpose of the Study:
- To investigate the impact of TQ-treated MSC conditioned medium (TQ-MSC-CM) on the biological characteristics of MCF7 breast cancer cells.
- To evaluate the effects of TQ-MSC-CM on reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP), cell death, cell cycle, and migration in MCF7 cells.
- To analyze the modulation of specific gene expressions (Cdk4, Sox2, c-Met, Bcl2) by TQ-MSC-CM in breast cancer cells.
Main Methods:
- MSCs were cultured and treated with TQ for 24 hours.
- Conditioned media from TQ-treated MSCs (TQ-MSC-CM) and control MSCs (MSC-CM) were collected.
- MCF7 cells were treated with TQ-MSC-CM and MSC-CM, followed by assays for ROS, MMP, apoptosis/necrosis, cell cycle, migration, and gene expression analysis (real-time PCR).
Main Results:
- TQ-MSC-CM and MSC-CM induced necrosis but not significant apoptosis in MCF7 cells.
- TQ-MSC-CM increased ROS production, decreased MMP, inhibited cell cycle progression, and downregulated Cdk4 and Sox2 gene expression.
- TQ-MSC-CM promoted MCF7 cell migration independently of c-Met expression and induced Bcl2 expression and Caspase-3/7 activity.
Conclusions:
- TQ-MSC-CM may induce programmed necrosis and inhibit proliferation and migration of breast cancer cells.
- The effects are mediated by modulating the paracrine activity of MSCs, suggesting TQ's potential in breast cancer treatment.
- TQ influences breast cancer cell behavior through mechanisms involving ROS, MMP, cell cycle regulation, and specific gene expression changes.
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