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Effect of toll-like receptor 3 agonists on the functionality and metastatic properties of breast cancer cell model
Nastaran Alizadeh1, Mohammad Mehdi Amiri, Alireza Salek Moghadam
1Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. khorami@yahoo.com.
Abstract:
There exists compelling evidence that Toll-like receptor 3 (TLR3) agonists can directly affect human cancer cells. The aim of this study was to investigate anti-cancer effects of TLR3 agonist in human breast cell line. We assessed potential effects of poly (A:U) on human breast cell line (MDA-MB-231) on a dose-response and time-course basis. Human breast cell line MDA-MB-231 was treated with different concentrations of poly (A:U) and lipopolysaccharide (LPS). Then, the following assays were performed on the treated cells: dose-response and time-course cytotoxicity using colorimetric method; matrix metalloproteinase-2 (MMP-2) activity using gelatin zymography method; apoptosis using annexin-v flowcytometry method; and relative expression of TLR3 and MMP-2 mRNA using reverse transcriptase polymerase chain reaction (RT-PCR) method. Following treatments, dose- response and time-course cytotoxicity using a colorimetric method, (MMP-2) activity (using gelatin zymography), apoptosis (using annexin-v flowcytometry method) assays and expression of TLR3 and MMP-2 genes (using PCR method) were performed. Cytotoxicity and flowcytometry analysis of poly (A:U) showed that poly (A:U) do not have any cytotoxic and apoptotic effects in different concentrations used. MMP-2 activity analysis showed significant decrease in higher concentrations (50 and 100 μg/ ml) between treated and untreated cells. Moreover, poly A:U treated cells demonstrated decreased expression of MMP-2 gene in higher concentrations. Collectively, our data indicated that human breast cancer cell line (MDA-MB-231) was highly responsive to poly (A:U). The antimetastatic effect of direct poly (A:U) and TLR3 interactions in MDA-MB-231 cells could provide new approaches in malignant tumor therapeutic strategy.
Insights
Toll-like receptor 3 (TLR3) agonists, like poly (A:U), show potential in targeting breast cancer. Poly (A:U) reduced MMP-2 activity and gene expression in MDA-MB-231 cells, suggesting an antimetastatic strategy.
Area of Science:
- Immunology
- Oncology
- Molecular Biology
Background:
- Toll-like receptor 3 (TLR3) agonists have demonstrated direct effects on human cancer cells.
- Investigating novel therapeutic strategies for breast cancer is crucial due to its prevalence.
Purpose of the Study:
- To evaluate the anti-cancer effects of a TLR3 agonist, poly (A:U), on the human breast cancer cell line MDA-MB-231.
- To assess the impact of poly (A:U) on cytotoxicity, apoptosis, matrix metalloproteinase-2 (MMP-2) activity, and gene expression.
Main Methods:
- Human breast cancer cells (MDA-MB-231) were treated with varying concentrations of poly (A:U).
- Assays included dose-response and time-course cytotoxicity, gelatin zymography for MMP-2 activity, annexin-v flow cytometry for apoptosis, and RT-PCR for TLR3 and MMP-2 gene expression.
Main Results:
- Poly (A:U) did not exhibit cytotoxic or apoptotic effects on MDA-MB-231 cells at the tested concentrations.
- A significant decrease in MMP-2 activity and MMP-2 gene expression was observed at higher concentrations of poly (A:U).
Conclusions:
- The human breast cancer cell line MDA-MB-231 responded effectively to poly (A:U).
- Direct interaction of poly (A:U) with TLR3 may offer an antimetastatic effect, presenting a potential therapeutic strategy for malignant tumors.
