Maryam Tohidast
1,2 
, Mohammad Amini
2, Mohammad Amin Doustvandi
2, Seyed Samad Hosseini
2 , Farzaneh Bilan
2, Nazila Mozammel
2, Pouryia Sameti
2, Amir Ali Mokhtarzadeh
2* , Behzad Baradaran
2* 1 Department of Biotechnology, Higher Education Institute of Rab-Rashid, Tabriz, Iran
2 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Abstract
Introduction: Colorectal cancer (CRC) is regarded as a serious global issue and is presently ranked second in the classification of gastrointestinal (GI) malignancies, with fast incidence and high mortality patterns. As the key “gene expression regulators”, miRNAs critically contribute to tumor progression and development. For example, miR-21 (an oncomiR) and miR-143 (a tumor suppressor) are dysregulated through colorectal tumorigenesis. Accordingly, this study assesses the concomitant therapeutic impacts of “miR-21 suppression” (anti-miR-21) and “miR-143 restoration” (miR-143) on CRC cell proliferation and migration.
Methods: SW-480 cell lines (with overexpressed “miR-21” and downregulated “miR-143”) were transfected via “anti-miR-21” and “miR-143” mimics, either independently or in combination. Next, cell viability assessment was performed through MTT assay. Then, apoptosis induction was examined with “Annexin V-FITC Kit”, and via Propidium Iodide (PI) assay and DAPI staining. In the next step, “cell cycle condition” and “autophagy induction” were studied through flow cytometry. “Wound-healing assay” and “clonogenic assay” were employed to investigate the migration and proliferation of tumor cells. Ultimately, qRT-PCR was utilized to quantify the intensity of the effects of “anti-miR-21” and “miR-143” on gene expression profiles.
Results: Downregulation of “miR-21” expression and overexpression of “miR-143” were found to synergistically reduce the viability (while elevating apoptosis) of SW-480 cells by modulating Bcl-2 and Bax expression profiles. Combined therapy increased the number of cells in the sub-G1 phase and reduced cell proliferation by modulating expression levels of PTEN and AKT-1. Additionally, miR-21 suppression and miR-143 restoration concomitantly reduced cell migration by modulating the expression of MMP-9.
Conclusion: Considering anti-cancer effects on cell growth, survival, and migration, it can be concluded that the concomitant suppression of “anti-miR-21” and “miR-143 restoration” might be introduced as a promising method for the therapy of CRC.