Investigation of coexpression correlation miR-۴۲۱ and miR-۲۱ in glioblastoma cell lines using bioinformatic and laboratory methods
- سال انتشار: 1403
- محل انتشار: دومین کنگره بین المللی کنسرژنومیکس
- کد COI اختصاصی: ICGCS02_464
- زبان مقاله: انگلیسی
- تعداد مشاهده: 102
نویسندگان
Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
چکیده
Introduction Glioblastoma (GBM) is the most aggressive glioma of astrocytic origin and is classified as grade ۴ according to the WHO classification. In the past years, studies have been conducted that show 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-۴۲۱ 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-۴۲۱ co-expression was performed. Also, Ago CLIP-seq data was analyzed. to investigate the expression correlation between miR-۲۱ and miR-۴۲۱, Pri miR-۴۲۱ was designed and cloned in to the expression vector. This vector transfected in to the U۸۷-MG cell line. 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 M-MLV kit. Then the expression level of miR-۲۱ and miR-۴۲۱ genes was measured using qRT-PCR. Also, the expression level of TIMP۳, RECK, SMAD۷ (related genes downstream of miR-۲۱ regulatory pathway) was measured by qRT-PCR. Results Bioinformatics analysis showed that miR-۴۲۱ and miR-۲۱ have co-expression with each other. The qRT-PCR results also showed that with the increase in the expression of miR-۴۲۱ in U۸۷-MG cell line, the expression of miR-۲۱ decreases and also the expression of TIMP۳ , SMAD۷, RECK genes increases. Conclusion Based on the analysis and the results we obtained, miR-۴۲۱, 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-۴۲۱, miR-۲۱, Glioblastomaمقالات مرتبط جدید
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