FUNCTIONAL ANALYSIS OF MIRNAS IN CERVICAL CANCER CELL LINES

Background: Genetics and epigenetics changes, have been showed as crucial regulators to cervical cancer progression. However, the mechanisms and role of miRNA dysregulation in cervical carcinogenesis are still not clear. We aimed to evaluate four miRNAs (miR-130a-3p, miR-205-5p, miR-4531 and miR-381-3p) and their targets regulation in human cervical cancer (CC) cell lines. Methods: We used a panel of CC cell lines including, five commercial cell line (HeLa, SiHa, CasKi, C4-I and C33-A) and one primary cell line (HCB-514) established in our lab. Primary Epidermal Keratinocytes cell line (HaCaT) was used as control. Differential expression level of miRNAs was assessed by quantitative PCR analysis. Gene expression profile was performed using nCounter PanCancer Pathways panel (NanoString Technologies®) to compare HCB-514 and HaCaT cell lines. The interaction networks of miRNA-target were constructed by Cytoscape program and REACTOME FI plug-in with CC category. Results: We found miR-130a-3p has been seen to be overexpressed in all CC cell lines, and miR-205-5p it was upregulated in CasKi, C4-IA, C33-A and HCB-514, both with a fold-change ≤2. However, miR-4531 showed no difference between expression levels in CC cell lines compared to HaCaT. On the other hand, miR-381-3p was downregulated only in HCB-514 cell line. Moreover, we identified 14 differentially expressed genes, eight downregulated (WNT10A, ITGA2, BMP7, DLL1, PPP2R2B, SOX9, BMP4 and DUSP6) and six upregulated (LTBP1, TMPRSS2, RASAL1, PAX8, IL6R and TLRB2) in HCB-514 when compared to HaCaT cell line. In the miRNAtarget interaction analysis, we verified that WNT10A can be being silenced by miR-130a-3p and miR-205-5p. Conclusion: This study revealed that miR-130a-3p and miR205-5p are upregulated in CC cell lines. WNT expression was predicted as a specific target of both miRNAs upregulated (miR-130a-3p and miR-205-5p). These results may be useful to better understanding of CC progression and the role of these microRNAs in this process.