Abstract
Purpose: Gastric cancer (GC) is one of the most frequent tumors with high mortality rate, worldwide. A proper understanding of the mechanism underlying its progression is required for its diagnosis and development of novel treatment option. MicroRNAs are associated with the development and advancement of different types of cancer, including GC. The current research was aimed at investigating the molecular and biological function of miR-148a-3p in GC development.Methods: A human normal gastric epithelial cell line, GES-1 (control) as well as four GC cell lines (NUGC-4, SNU-520, STKM-2 and MKN-74) were employed for the study. MiR-148a-3p and ATP6AP2 expression levels in GC cell lines were examined by RT-qPCR technique. Transfection procedure was used to upregulate miR-148a-3p expression in the MKN-45 cell line. MTT assay was utilized to evaluate cell viability in GC cell lines. The molecular interaction between miR-148a-3p and ATP6AP2 was predicted using bioinformatics system and the prediction was then validated by luciferase reporter assay.Results: Expression levels of miR-148-3p was low, whilst that of ATP6AP2 was high in GC cell lines. MiR-148a-3p overexpression resulted in the reduction of cell viability in GC cell lines. More so, it was confirmed that miR-148-3p, as a post-transcriptional regulator inhibited ATP6AP2 expression by having a negative association with it in GC cells. More so, ATP6AP2 was found to be a direct target of miR-148a-3p.Conclusion: Our results revealed that miR-148a-3p plays a crucial function in GC development through targeting ATP6AP2. This finding could be explored in the discovery of new therapeutic approaches for GC treatment.
 Keywords: ATP6AP2, Cell viability, Gastric cancer, miR-148a-3p, Progression
Highlights
Gastric cancer (GC) is recorded to be the number five most frequent tumor and the number three chief source of mortalities, caused by cancers worldwide [1]
More researches proposed that the functions played by miRNA in the carcinoma progression include modulating growth of tumor, metastasis, as well as epithelial–mesenchymal transition (EMT)
MiR-148a-3p is downregulated in gastric cancer cells
Summary
Gastric cancer (GC) is recorded to be the number five most frequent tumor and the number three chief source of mortalities, caused by cancers worldwide [1]. Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, Thermo Fisher Scientific, New Jersey, USA) containing 5 percent fetal bovine serum (fbs; ABCAM, San Francisco, United States), 5 percent CO2 and 95 percent air in a moistened incubator (Beyotime Institute of Biotechnology, Beijing, China) were used for culturing of the cells. The 2-DDCt system was used to analyse the ultimate comparative quantities of target mRNAs and miRNAs. RNA overexpression plasmid was intended to precisely target ATP6AP2 by means of oe-RNA inventing apparatuses (ABCAM, San Francisco, United States). The miR-148a-3p mimics was produced by Ambion (Invitrogen, ABCAM, San Francisco, United States) and transfected into MKN-45 cell to a last solution of 100 nM using Lipofectamine 2000 (Invitrogen; Beyotime Institute of Biotechnology, Haimen, China) for 2 days at 37 oC, following the instructions of the manufacturer. Student’s t-test was used to examine the differences between two groups; the one-way analysis of variance was used to analyze the inter-group variances, a post hoc Tukey test for numerous contrasts
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