Effect of Toll-Like Receptor 3 Agonists on the Functionality and Metastatic Properties of Breast Cancer Cell Model

• Main Authors: Nastaran Alizadeh, Mohammad Mehdi Amiri, Alireza Salek Moghadam, Amir Hassan Zarnani, Farshid Saadat, Farnaz Safavifar, Azar Berahmeh, Mohammad Reza Khorramizadeh
• Format: Article
• Language: English
• Published: Tehran University of Medical Sciences 2013-06-01
• Series: Iranian Journal of Allergy, Asthma and Immunology
• Subjects: MDA-MB-231; MMP-2; Poly (A; U); TLR3
• Online Access: https://ijaai.tums.ac.ir/index.php/ijaai/article/view/518
• ISSN: 1735-1502; 1735-5249

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.