title: Effect of toll-like receptor 3 agonists on the functionality and metastatic properties of breast cancer cell model.
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.
authors.affiliation: Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. khorami@yahoo.com.
authors.forename: Nastaran
authors.initials: N
authors.lastname: Alizadeh
authors.forename: Mohammad Mehdi
authors.initials: MM
authors.lastname: Amiri
authors.forename: Alireza
authors.initials: A
authors.lastname: Salek Moghadam
authors.forename: Amir Hassan
authors.initials: AH
authors.lastname: Zarnani
authors.forename: Farshid
authors.initials: F
authors.lastname: Saadat
authors.forename: Farnaz
authors.initials: F
authors.lastname: Safavifar
authors.forename: Azar
authors.initials: A
authors.lastname: Berahmeh
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Khorramizadeh
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001943:Breast Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D011063:Poly A-U / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051196:Toll-Like Receptor 3 / Q000819:agonists*
chemical_list: C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D011063:Poly A-U; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2