MicroRNA-567 inhibits cell proliferation and induces cell apoptosis in A549 NSCLC cells by regulating cyclin-dependent kinase 8
| Autor: | Ahmed S. Doghish, Ahmed Elshafei, Mostafa M. Elshafey, Mohamed A. Elkady |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
FITC
Fluorescein isothiocyanate 0106 biological sciences 0301 basic medicine MiR-567 Cell cycle checkpoint MiR or MiRNA MicroRNA CDK8 Apoptosis Cell cycle NSCLC 01 natural sciences DMEM Dulbecco’s modified Eagle’s medium DAPI 4′ 6-Diamidino-2 Phenylindole Dihydrochloride 03 medical and health sciences PBS phosphate buffer saline FBS fetal bovine serum Downregulation and upregulation RIPA Radio immunoprecipitation assay microRNA LC Lung cancer NSCLC Non-small cell lung cancer lcsh:QH301-705.5 Cell proliferation A549 cell h Hour Chemistry Cell growth ATCC American type culture collection DMSO Dimethyl sulfoxide Transfection PI Propidium iodide PVDF Poly-vinylidene fluoride 16HBE Normal human bronchial epithelial cell line mRNA Messenger RNA respiratory tract diseases 030104 developmental biology lcsh:Biology (General) cDNA Complementary DNA Cancer research Original Article BrdU 5-bromo- 2′-deoxyuridine General Agricultural and Biological Sciences MTT 3-(4 5-dimethylthiazol-2-yl)-2 5 diphenyltetrazolium bromide qRT-PCR Quantitative real time-polymerase chain reaction CDK8 Cyclin-dependent kinase 8 010606 plant biology & botany |
| Zdroj: | Saudi Journal of Biological Sciences, Vol 28, Iss 4, Pp 2581-2590 (2021) Saudi Journal of Biological Sciences |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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