Open AccessEvaluation of Silencing Effect of miR-133a-5p Mimic on TIM-3 Expression in AML (HL-60) Cell Line
Author(s) -
Zohreh Hojati,
Mazdak Ganjalıkhani-Hakemi,
Mahnaz Ameri,
Seyedeh Fatemeh Alimohammadi-Jelodar,
Moein Dehbashi,
Maryam Mohammad Ganji,
Vida Homayouni,
Hossein Khanahmad
Publication year - 2019
Publication title -
indian journal of clinical biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.392
H-Index - 42
ISSN - 0970-1915
DOI - 10.1007/s12291-019-00834-z
Subject(s) - gene silencing , microrna , transfection , flow cytometry , biology , cell culture , microbiology and biotechnology , cancer research , gene , genetics
Acute myelogenous leukemia (AML) is a complex blood malignancy leading to immature leukemic stem cells (LSCs) proliferation. T cell immunoglobulin mucin-3 (TIM-3) is known as a biomarker of AML LSCs. Several microRNAs (miRNAs) can affect gene expression in AML. In this study, the silencing effect of miR-133a-5p on TIM-3 expression in AML cell lineage (HL-60) was investigated. It's been hypothesized that miR-133a-5p may suppress the TIM-3 expression in AML cell line. Initially, miRNA-TIM-3 prediction, enrichment, and network analysis were done. Then, miR-133a-5p mimic was transfected into HL-60 cells. The TIM-3 protein and gene expression were measured by flow cytometry analysis and real-time PCR, respectively. MTT assay was also carried out. Based on the Bioinformatics predictions, miR-133a-5p was able to silence TIM-3 expression. Also, significant pathways pertained to miR-133a-5p were obtained using enrichment analysis. According to this, miR-133a-5p was mainly engaged in the MAPK signaling pathway and Nicotine addiction pathway using the KEGG database. The TIM-3 protein expression of the transfected cells was measured as 17.15 ± 8.87% ( p = 0.001). A 52.48% significant gene silencing in mRNA level was obtained in comparison to the negative control. Despite of down regulation of TIM-3, HL-60 cell viability has not been significantly changed. It has been finally confirmed that miR-133a-5p could strongly suppress TIM-3 expression in AML cell line. Presumably, down regulation of TIM-3 could affect MAPK and Nicotine addiction signaling pathways.