Zahra Sadat Hashemi - Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
mehdi Forouzandeh Moghadam - Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Samila Farokhimanesh - Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
masuomeh rajabibazl - Department of Clinical Biochemistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Esmaeil sadroddiny - Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran

Objective(s): Various studies have been conducted to reduce the metastatic behavior of cancerous cells. In this regard, ectopic expression of anti-metastatic microRNAs by miR-mimic and miR-restoration-based therapies could bring new insights to the field. In the present study, the consequences of co-transfecting breast cancer cell lines with miR-۱۹۳b and miR-۳۱ were investigated via invasion and migration assays.Materials and Methods: Double stranded oligonucleotide of mature miR-۱۹۳b-۳p and miR-۳۱-۵p were cloned into pcDNA ۶.۲gw/EmGFP plasmid. The resulting plasmids were used for transfection. Real time-PCR was performed to assess the expression of miR-۱۹۳b and miR-۳۱ as well as Ras homolog gene family member A (RhoA) and urokinase-type plasminogen activator (uPA) as miR targets. Scratch, Transwell migration and Matrigel invasion assays were carried out to assess the extent of migration and invasion of cell lines.Results: The most significant increase in expression of miRs belonged to the single transfection of mimic-miRs in MDA-MB۲۳۱. Although the co-transfection was not as successful as single transfection in miR expression, it was significantly more effective in inhibition of the cells invasive potential. Conclusion: Although the miR-restoration therapy based on co-transfection of two miRs could be less effective in expression of each miRNA, the resulting decrease in metastatic behavior of the cells is more significant due to collective effect of co-transfection to decrease target gene expression. Our results revealed that employing this sort of combinatorial strategies could lead to more efficient reduction in metastatic behavior. It seems that using this strategy would bring about more successful therapeutic outcomes.

Breast Cancer, Metastasis, miR-۳۱, miR-۱۹۳b, RhoA, uPA