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Bioimpacts. 2024;14(3): 29913.
doi: 10.34172/bi.2023.29913
  Abstract View: 125
  PDF Download: 89

Original Article

Cooperatively inhibition effect of miR-143-5p and miR-145-5p in tumorigenesis of glioblastoma cells through modulating AKT signaling pathway

Sheyda Jodeiry Zaer 1,2 ORCID logo, Mahmoudreza Aghamaali 1*, Mohammad Amini 2, Mohammad Amin Doustvandi 2, Seyed Samad Hosseini 2 ORCID logo, Behzad Baradaran 2, Souzan Najafi 2, Yalda Baghay Esfandyari 2, Ahad Mokhtarzadeh 2* ORCID logo

1 Department of Biology, Faculty of Sciences, University of Guilan, Rasht, Iran
2 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
*Corresponding Authors: Mahmoudreza Aghamaali, Email: aghamaali@guilan.ac.ir; Ahad Mokhtarzadeh, Mokhtarzadehah@tbzmed.ac.ir, Email: Ahad.mokhtarzadeh@gmail.com

Abstract

Introduction: As the most common aggressive primary brain tumor, glioblastoma is inevitably a recurrent malignancy whose patients’ prognosis is poor. miR-143 and miR-145, as tumor suppressor miRNAs, are downregulated through tumorigenesis of multiple human cancers, including glioblastoma. These two miRNAs regulate numerous cellular processes, such as proliferation and migration. This research was intended to explore the simultaneous replacement effect of miR-143, and miR-145 on in vitro tumorgenicity of U87 glioblastoma cells.
Methods: U87 cells were cultured, and transfected with miR-143-5p and miR-145-5p. Afterward, the changes in cell viability, and apoptosis induction were determined by MTT assay and Annexin V/PI staining. The accumulation of cells at the cell cycle phases was assessed using the flow cytometry. Wound healing and colony formation assays were performed to study cell migration. qRT-PCR and western blot techniques were utilized to quantify gene expression levels.
Results: Our results showed that miR-143-5p and 145-5p exogenous upregulation cooperatively diminished cell viability, and enhanced U-87 cell apoptosis by modulating Caspase-3/8/9, Bax, and Bcl-2 protein expression. The combination therapy increased accumulation of cells at the sub-G1 phase by modulating CDK1, Cyclin D1, and P53 protein expression. miR-143/145-5p significantly decreased cell migration, and reduced colony formation ability by the downregulation of c-Myc and CD44 gene expression. Furthermore, the results showed the combination therapy of these miRNAs could remarkably downregulate phosphorylated-AKT expression levels.
Conclusion: In conclusion, miR-143 and miR-145 were indicated to show cooperative anti- cancer effects on glioblastoma cells via modulating AKT signaling as a new therapeutic approach.
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Submitted: 18 May 2023
Revision: 30 Sep 2023
Accepted: 07 Oct 2023
ePublished: 05 Nov 2023
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