Identification and validation of putative target genes regulated by miR-34 in cervical cancer

PurposeWe aimed to identify the epithelial-mesenchymal transition and associated putative targets of the miR-34 family in transcriptomic data of cervical epithelial squamous cell carcinoma (CESC) to find new therapeutic targets for better disease management. MethodsA combined computational analysis of the miR-34 family; gene expression in heterogeneous primary CESCs derived from TCGA; and the integration of miR-34b and EMT-regulated genes was performed. Four EMT-associated miR-34b gene targets were analysed in primary human CESC and non-cancerous cervical tissues by qRT-PCR. Effects of endogenous miR-34b expression and its associated gene modulations in cervical cancer cells (C33A and HeLa) were analysed using qRT-PCR, western blotting and immunofluorescence, transwell migration and invasion assays. ResultsThe results showed that the miR-34 family might regulate the mTOR pathway, cell cycle (CCND2) and cell adhesion functions (FZD4). Further, we showed that a low negative correlation (r2= -0.07) between miR-34b/EM[I] score and four EMT signature genes (E>MP7, CAV1, ID2, FN1) were significantly regulated by miR-34b. Also, FN1 was indicated as its putative target with the highest negative EMT score and high binding energy between MRE/3UTR. Further, these transcriptomic signatures in CESC revealed a significant inverse correlation across the stages of primary human CESC. These genes were repressed at transcriptional and translational levels in miR-34b-3p expressing C33A and HeLa cells, contributing to their reduced cell migration and invasive properties. ConclusionsOur studies revealed the potential targets of the miR-34 family, especially miR-34b, that can emerge as potential biomarkers and promising therapeutic targets in CESC disease management.