1. Regulatory Effect of let-7f Transfection in Non-Small Cell Lung Cancer on its Candidate Target Genes
- Author
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Zafari V., Asadi M., Bakhtiyari N., Sadeghzadeh M., Khalili M., Zarredar H., and Bornehdeli S.
- Subjects
gene overexpression ,high mobility group A2 protein ,cancer cell culture ,cancer prognosis ,HMGA2 gene ,Article ,gene targeting ,Western blotting ,let 7f gene ,male ,molecular pathology ,middle aged ,controlled study ,human ,tumor suppressor gene ,Non-Small-Cell Lung ,FZD3 gene ,protein expression ,cell viability ,clinical article ,MTT assay ,non small cell lung cancer ,ARID3B gene ,quantitative analysis ,adult ,human cell ,Carcinoma ,bioinformatics ,gene control ,human tissue ,MicroRNAs ,cell proliferation ,female ,real time polymerase chain reaction ,A-549 cell line ,tumor marker ,genetic transfection ,SMARCAD1 gene ,Biomarkers ,cancer tissue ,gene expression level - Abstract
Background: Let-7f has essential impacts on biological processes; however, its biological and molecular functions in lung cancer pathogenesis have yet been remained unclear. We aimed to investigate the expression level of let-7f and its candidate target genes both in lung cancer tissues and A549 cell line. Methods: Bioinformatics databases were first used to select candidate target genes of let-7f. Then the relative gene and protein expressions of let-7f and its target genes, including HMGA2, ARID3B, SMARCAD1, and FZD3, were measured in lung tissues of NSCLC patients and A549 cell line using qRT-PCR and Western blotting. The electroporation method was used to transfect A549 cells with let-7f mimic and microRNA inhibitor. The impact of let-7f transfection on the viability of A549 cells was assessed using MTT assay. The expression data of studied genes were analyzed statistically Results: Results indicated significant downregulated expression level of let-7f-5p (p = 0.0013) and upregulated level of the HMGA2 and FZD3 in NSCLC cases (p < 0.05). In A549 cells, after transfection with let-7f mimic, the expression of both mRNA and protein levels of HMGA2, ARID3B, SMARCAD1, and FZD3 decreased. Also, the overexpression of let-7f significantly inhibited the A549 cell proliferation and viability (p = 0.017). Conclusion: Our findings exhibited the high value of let-7f and HMGA2 as biomarkers for NSCLC. The let-7f, as a major tumor suppressor regulatory factor via direct targeting genes (e.g. HMGA2), inhibits lung cancer cell viability and proliferation and could serve as a marker for the early diagnostic of NSCLC. © 2022, Pasteur Institute of Iran. All rights reserved., Ministry of Health and Medical Education, MOHME; Tabriz University of Medical Sciences, TUOMS, This study was supported by the Non-Communicable Center of the Ministry of Health and Medical Education, and Tuberculosis and Lung Disease Center of Tabriz University of Medical Sciences (Tabriz, Iran. of Medical Sciences.
- Published
- 2022