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Copyright (c) 2024 Sara Ezzat, Magda I Mohamad, Manal Basyouni Ahmed, Sara Elsayed Abdelrahman, Nesma Hussein Abdel hay, Nashwa El-Khazragy, Maha Imam Ismail
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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.The tumor suppressor effect of miRNA 200-c-3p on A549 non-small cell lung cancer cell line is associated with down-regulation of programmed cell death ligand 1; an immune-check point mechanism
Corresponding Author(s) : Sara Ezzat
Cellular and Molecular Biology,
Vol. 70 No. 3: Issue 3
Abstract
MiRNA 200-c-3p has varying functions in different tumor types, whether tumor suppression or promotion. Comprehensive assessment of its function in non-small cell lung cancer (NSCLC) together with its effect on antitumor immune response have not been declared before. We aimed to explore the effect of replacement and suppression of miRNA 200-c-3p on (NSCLC) and its impact on immune checkpoint function and subsequently antitumor immunity. MiRNA 200-c-3p mimic/inhibitor was transfected into the A549 (NSCLC). A 549 non-small cell lung cancer cells viability was done by trypan blue staining and 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flowcytometric analysis was done for apoptosis detection. Real‐time reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used to study its effect on relative gene expression and relative protein level of programmed cell death ligand 1 (PD-L1). Finally, co-culture with isolated and activated T cells was performed. Multiple comparisons were performed using one-way analysis of variance (ANOVA) followed by Tukey’s multiple-comparison test. Decreased cell viability, increased apoptosis, reduced PD-L1 relative gene expression and its relative protein level, together with enhanced T cell cytotoxicity towards tumor cells were detected after miRNA 200-c-3p mimic transfection of A549 NSCLC cell line. However, these results were reversed in miRNA 200-c-3p suppression. MiRNA 200-c-3p had a tumor suppressive effect in non-small cell lung cancer cells which might be through down regulation of PD-L1 relative gene expression, and it may be used as a new target to improve immune checkpoint dysfunction.
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