Author/Authors :
Hashemi, Zahra Sadat Department of Medical Biotechnology - School of Advanced Technologies in Medicine - Tehran University of Medical Sciences, Tehran, Iran , Forouzandeh Moghadam, Mehdi Department of Medical Biotechnology - Faculty of Medical Sciences - Tarbiat Modares University, Tehran, Iran , Sadroddiny, Esmaeil Department of Medical Biotechnology - School of Advanced Technologies in Medicine - Tehran University of Medical Sciences, Tehran, Iran
Abstract :
Background: Metastasis-associated miRNA (metastamiR) harbors a great potential to confine metastasis as the most lethal aspect
of cancer. miR-193b-3p is an anti-metastatic miRNA, whose expression significantly decreases in metastatic breast cancer cells.
Objectives: In the present study, the expression patterns of different cell-lines were investigated, following an effective miR transfection
strategy.
Methods: Double-stranded oligo of mature miR-193b-3p, miR-negative, and miR-LacZ were designed and cloned into
pcDNA6.2gw/EmGFP plasmid. Calculating the population doubling time (PDT), non-tumorigenic MCF-10A, tumorigenic but
non-metastatic MCF-7, and metastatic MDA-MB231 were transfected by Lipofectamin2000 and Express-In. The expressions of
miR-193b-3p and miR-191 have been quantified by Real-time PCR 48 hours after transfection. Scratch, Transwell migration, and
Matrigel invasion assays have been carried out to assess the migration and invasion levels of the cell-lines.
Results: The PDT (21.270.43, 28.180.34, and 35.830.44 hours) and miR-193b-3p relative expression before transfection (100, 42
and 8) were measured for MCF-10A, MCF-7, and MDA-MB231 cell-lines, respectively. Better transfection results were obtained based on
nano-liposomemethod. Theexpression of the miR-193b-3pwasincreased 32, 19 and, 65 fold-change. Therate of invasion in metastatic
cells was 6.6 fold-higher in comparison to MCF-7 cells.
Conclusions: Higher expression rate of the miRNA is anticipated to occur, following miR-mimic transfection. However, the observed
differential patterns of miRNA increase in the context of different cell-lines, indicating the involvement of more complicated
cellular pathways. Scrutinizing these cellular mechanisms could open new horizons in cancer therapy and management strategies.