Overexpression of miR-23b-3b+miR-218-5p+miR-124-3p differentially modifies the transcriptome of C-33A and CaSki cells and the regulation of cellular processes involved in the progression of cervical cancer
Dysregulation of tumor suppressor miRNAs (tsmiRs) is associated with tumor progression in cancer. miR-23b-3p, miR-218-5p and miR-124-3p are tsmiRs in cervical cancer (CC) and regulate the translation of genes involved in metastasis-related biological processes. Objective. To analyze transcriptome changes in cervical cancer cell lines (C-33A HPV-negative and CaSki HPV-positive) overexpressing miR-23b-3b+miR-218-5p+miR-124-3p, to identify specific target transcripts common to all three miRNAs, as well as signaling pathways and cellular processes related to tumor progression. Methods. The transcriptome of C-33A and CaSki cells transfected with miR-23b-3b+miR-218-5p+miR-124-3p was analyzed by RNA-seq. Differentially expressed genes (DEGs) were subjected to Gene Ontology analysis on the DAVID platform. The function of under-regulated genes was analyzed on the GEPIA 2.0, Kaplan-Meier plotter and STRING platforms. On the TargetScanHuman platform it was determined which transcripts have MREs for miR-23b-3p, miR-218-5p and/or miR-124-3p in their 3`UTR region. Results. Simultaneous overexpression of miR-218-5p, miR-124-3p and miR-23b-3p induced changes in global gene expression in C-33A and CaSki cells. In C-33A cells, DEGs included 45 over- and 172 under-regulated transcripts; in CaSki, 125 transcripts were over- and 84 under-regulated. The under-regulated transcripts enrich proliferation, migration, apoptosis and angiogenesis; 20 of these genes are associated with overall survival (OS) in women with CC, and 18 of the 20 mRNAs have MREs for one, two or all three miRNAs. Conclusions. miR-23b-3b+miR-218-5p+miR-124-3p, differentially modify global gene expression in C-33A and CaSki cells. The results indicate that these miRNAs act synergistically and modulate CC progression through individual and shared targets by two or all three miRNAs. Overall design: We analyzed the effect of miR-23b-3b+miR-218-5p+miR-124-3p overexpression on the transcriptome of C-33A and CaSki cells, with the aim of identifying differential expression profiles between cell lines, recognizing specific target transcripts shared by the three miRNAs, as well as gene sets involved in signaling pathways and cellular processes related to cervical cancer progression. C-33A and CaSki cells were transfected with mimic miRNAs: hsa-miR-23b-3p; hsa-miR-124-3p; and hsa-miR-218-5p. As a negative control mirVana™ miRNA Mimic, Negative Control #1 (scramble). To analyze changes in the transcriptome of C-33A and CaSki cells overexpressing miR-23b-3b+miR-218-5p+miR-124-3p, to identify specific target transcripts common to all three miRNAs, the transcriptome of C-33A and CaSki cells was analyzed by RNA-seq. Three independent experiments were performed for each condition to ensure reproducibility.