Bioinformatics and laboratory analysis of miR-۵۰۵ and miR-۲۱ expression correlation in glioblastoma

Introduction In recent years, studies have shown that non-coding RNAs play an important role in cancer. An important group of these ncRNAs are miRs, which are important regulators in the cell. miR-۲۱ is one type of miR that plays an oncogenic role in various cancers, especially GBM. On the other hand, according to the studies, miR-۵۰۵ is known as a tumor suppressor (TSG) and is significantly decreased in GBM tissues and cell lines. Studies conducted in recent years indicate that miRs may target other ncRNAs, including other miRs. This mode is called miR: miR interaction. Methods At first, bioinformatics analysis of miR-۲۱ and miR-۵۰۵ co-expression was performed. Also, Ago CLIP-seq data was analyzed to investigate the miR: miR Interactions between miR-۲۱ and miR-۵۰۵. For laboratory analysis, Pri miR-۵۰۵ was designed and cloned into the expression vector. This vector was transfected into the U۸۷-MG cell line. Also, the transfected state of the mock vector and the non-transfected were considered in this experiment. Total RNA was extracted, and DNase treatment was performed on the extracted RNA using a DNase I, RNase-free kit. Next ۱ug of total RNA was used to synthesize cDNA with an M-MLV kit. Then the expression level of miR-۲۱ and miR-۵۰۵ genes was measured using qRT-PCR. Also, the expression level of PDCD۴, HNRNPK, SMAD۷, RECK, and HPRTII (related genes downstream of the miR-۲۱ regulatory pathway) was measured by qRT-PCR. Results Bioinformatics analysis showed that miR-۵۰۵ and miR-۲۱ correlate with each other. The qRT-PCR results also showed that with the increase in the expression of miR-۵۰۵ in cells, the expression of miR-۲۱ decreases, and the expression of PDCD۴, HNRNPK, SMAD۷, RECK and HPRTII increases significantly. Conclusion Based on the analysis and the results we obtained, miR-۵۰۵, by targeting miR-۲۱, can change the inhibition of TSGs targeted by miR-۲۱ and cause their expression to return in GBM, which inhibits GBM tumor cells.

کلیدواژه ها:

miR-۲۱ ، miR-۵۰۵ ، Glioblastoma