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微小RNA-567通过调节细胞周期蛋白依赖性激酶8抑制A549非小细胞肺癌细胞的增殖并诱导其凋亡。

MicroRNA-567 inhibits cell proliferation and induces cell apoptosis in A549 NSCLC cells by regulating cyclin-dependent kinase 8.

作者信息

Elkady Mohamed A, Doghish Ahmed S, Elshafei Ahmed, Elshafey Mostafa M

机构信息

Department of Biochemistry and Molecular Biology, Faculty of Pharmacy (Boys), Al-Azhar University, Nasr City, Cairo 11651, Egypt.

Department of Biochemistry, Faculty of Pharmacy, Badr University in Cairo (BUC), Badr City, Cairo 11829, Egypt.

出版信息

Saudi J Biol Sci. 2021 Apr;28(4):2581-2590. doi: 10.1016/j.sjbs.2021.02.001. Epub 2021 Feb 14.

Abstract

MicroRNA-567 (miR-567) plays a decisive role in cancers whereas its role in non-small cell lung cancer (NSCLC) is still unexplored. This study was therefore planned to explore the regulatory function of miR-567 in A549 NSCLC cells and investigate its possible molecular mechanism that may help in NSCLC treatment. In the current study, miR-567 expression was examined by quantitative real time-polymerase chain reaction (qRT-PCR) in different NSCLC cell lines in addition to normal cell line. A549 NSCLC cells were transfected by miR-567 mimic, miR-567 inhibitor, and negative control siRNA. Cell proliferation was evaluated by MTT and 5-bromo-2'deoxyuridine assays. Cell cycle distribution and apoptosis were studied by flow cytometry. Bioinformatics analysis programs were used to expect the putative target of miR-567. The expression of cyclin-dependent kinase 8 (CDK8) gene at mRNA and protein levels were evaluated by using qRT-PCR and western blotting. Our results found that miR-567 expressions decreased in all the studied NSCLC cells as compared to the normal cell line. A549 cell proliferation was suppressed by miR-567 upregulation while cell apoptosis was promoted. Also, miR-567 upregulation induced cell cycle arrest at sub-G1 and S phases. CDK8 was expected as a target gene of miR-567. MiR-567 upregulation decreased CDK8 mRNA and protein expression while the downregulation of miR-567 increased CDK8 gene expression. These findings revealed that miR-567 may be a tumor suppressor in A549 NSCLC cells through regulating CDK8 gene expression and may serve as a novel therapeutic target for NSCLC treatment.

摘要

微小RNA-567(miR-567)在癌症中起决定性作用,但其在非小细胞肺癌(NSCLC)中的作用仍未被探索。因此,本研究旨在探讨miR-567在A549非小细胞肺癌细胞中的调控功能,并研究其可能有助于非小细胞肺癌治疗的分子机制。在本研究中,除了正常细胞系外,还通过定量实时聚合酶链反应(qRT-PCR)检测了不同非小细胞肺癌细胞系中miR-567的表达。用miR-567模拟物、miR-567抑制剂和阴性对照小干扰RNA转染A549非小细胞肺癌细胞。通过MTT和5-溴-2'-脱氧尿苷试验评估细胞增殖。通过流式细胞术研究细胞周期分布和凋亡。使用生物信息学分析程序预测miR-567的假定靶标。通过qRT-PCR和蛋白质印迹法评估细胞周期蛋白依赖性激酶8(CDK8)基因在mRNA和蛋白质水平的表达。我们的结果发现,与正常细胞系相比,所有研究的非小细胞肺癌细胞中miR-567的表达均降低。miR-567上调抑制A549细胞增殖,同时促进细胞凋亡。此外,miR-567上调诱导细胞周期停滞在亚G1期和S期。CDK8被预测为miR-567的靶基因。miR-567上调降低CDK8 mRNA和蛋白质表达,而miR-567下调增加CDK8基因表达。这些发现表明,miR-567可能通过调节CDK8基因表达在A549非小细胞肺癌细胞中发挥肿瘤抑制作用,并可能成为非小细胞肺癌治疗的新靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/587a/8071907/076b8cbdfa6f/gr1.jpg

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