Influence of hsa-miR-1265 and hsa-miR-21-5p on meningioma cells
Meningiomas are common intracranial tumors. Most of thenm benign WHO GI tumors, while approximately 20% are diagnosed as progressively more aggressive GII and GIII meningiomas. The study aimed to identify genes with tumor grade-related expression and to assess their functional relevance. The effect of selected miRNAs including hsa-miR-21-5p and hsa-miR-1265 on the cell phenotypep and genes expression profile was investigated using in vitro culturing of menignigioma cells KT21-MG1 andd Ben-Men-1. The cells were cultured with specific synthetic miRNA (miRNA mimic) or or nonspeciic control miRNA. Genes expression in miRNA-treated anad control cells was determined with RNAseq. Overall design: KT21-MG1 and Ben-Men-1 meningioma cells were transfected syntetic miRNA mimics using liposomal transfection. The following miRNA micmics were used hsa-miR-1265 (YM00470622-ADB , Qiagen), hsa-miR-21-5p (YM00470856-ADB, Qiagen) and negative control mimic (YM00479902-AGB; Qiagen) that serves as nonspecific negative control. Three independent biological replicates of the transfection experiment with each miRNA was performed and tptal RNA was isolated using High Pure RNA Isolation Kit (Roche). RNA sequencing was performed using poly-A enrichment or library constructing. One µg of RNA from each tissue sample was used for mRNA library preparation with Novogene NGS RNA Library Prep Set (Novogene). The quality of libraries was evaluated with Agilent Bioanalyzer 2100 system (Agilent Technologies). Libraries were sequenced on an Illumina NovaSeq 6000 platform, and 150-bp paired-end reads were generated. A minimum of 30 M read pairs per sample were generated. Sequencing was performed by Eurofins Genomics service.