miR­302c­3p and miR­520a­3p suppress the proliferation of cervical carcinoma cells by targeting CXCL8.

The aim of the present study was to identify the differentially expressed microRNAs (miRs) in cervical carcinoma (CC) tissues and cells and to explore the function of miR­302c­3p and miR­520a­3p in the proliferation of CC cells. Potential dysregulated miRNAs in CC tissues and tumour­adjacent tissues were detected. Reverse transcription­quantitative PCR (RT­qPCR) was performed to determine the expression of miR­302c­3p, miR­520a­3p and CXCL8 in CC tissues and cell lines. The target genes of the miRNAs were predicted using miRTarBase and verified by luciferase reporter assays. RT­qPCR and western blotting were performed to measure the expression of C­X­C motif ligand (CXCL)8 after transfection. The effect on proliferation was verified by Cell Counting Kit assay and ethynyl­2­deoxyuridine staining. Flow cytometry was utilised to assess the effect on apoptosis. In the present study, miR­302c­3p and miR­520a­3p were markedly downregulated in CC cell lines compared to the normal cervical cell line H8. Functionally, overexpression of miR­302c­3p and/or miR­520a­3p inhibited proliferation and promoted the apoptosis of CC cell lines in vitro, while the knockdown of miR­302c­3p and/or miR­520a­3p had the opposite effect. Furthermore, miR­302c­3p and miR­520a­3p could both bind to CXCL8. Inhibition of CXCL8 in combination with miR­302c­3p and/or miR­520a­3p overexpression exerted proliferation­suppressive and apoptosis­stimulating effects on CC cells, whereas restoring CXCL8 attenuated the miR­302c­3p­ and miR­520a­3p­induced anti­proliferative and pro­apoptotic effects. miR­302c­3p and miR­520a­3p suppress the proliferation of CC cells by downregulating the expression of CXCL8, which may provide a novel target for the treatment of CC.

Hsa_circRNA_102610 upregulation in Crohn's disease promotes transforming growth factor-ß1-induced epithelial-mesenchymal transition via sponging of hsa-miR-130a-3p.

BACKGROUND: The incidence of inflammatory bowel disease, a chronic intestinal inflammatory disorder that includes Crohn's disease (CD) and ulcerative colitis, is rising. Circular RNAs are considered valuable diagnostic biomarkers for CD. Current evidence supports the views that epithelial-mesenchymal transition (EMT) plays an important role in CD pathogenesis, and that hsa-miR-130a-3p can inhibit transforming growth factor-ß1 (TGF-ß1)-induced EMT. Our previous study revealed that hsa_circRNA_102610 was upregulated in CD patients. Moreover, we predicted an interaction between hsa_circRNA_102610 and hsa-miR-130a-3p. Thus, we hypothesized that hsa_circRNA_102610 may play roles in the proliferation and EMT of intestinal epithelial cells by sponging hsa-miR-130a-3p to participate in the pathogenesis of CD. AIM: To explore the mechanism of hsa_circRNA_102610 in the pathogenesis of CD. METHODS: The relative expression levels of hsa_circRNA_102610 and hsa-miR-130a-3p in patients were detected by quantitative reverse transcription-polymerase chain reaction. The proliferation of human intestinal epithelial cells (HIECs) and normal-derived colon mucosa cell line 460 (NCM460) cells was detected by cell counting kit-8, 5-ethynyl-2'-deoxyuridine staining and cell cycle assays following overexpression or downregulation of hsa_circRNA_102610. Cell proliferation assays were performed as described above in a rescue experiment with hsa-miR-130a-3p mimics. The interaction of hsa_circRNA_102610 and hsa-miR-130a-3p was verified by fluorescence in situ hybridization and dual luciferase reporter assays. The relative expression levels of CyclinD1, mothers against decapentaplegic homolog 4 (SMAD4), E-cadherin, N-cadherin and Vimentin were detected by western blotting following hsa_circRNA_102610 overexpression, TGF-ß1-induced EMT or hsa-miR-130a-3p mimic transfection (in rescue experiments). RESULTS: Upregulation of hsa_circRNA_102610 was determined to be positively correlated with elevated fecal calprotectin levels in CD (r = 0.359, P = 0.007) by Pearson correlation analysis. Hsa_circRNA_102610 promoted the proliferation of HIECs and NCM460 cells, while hsa-miR-130a-3p reversed the cell proliferation-promoting effects of hsa_circRNA_102610. Fluorescence in situ hybridization and dual luciferase reporter assays showed that hsa_circRNA_102610 directly bound hsa-miR-130a-3p in NCM460 and 293T cells. An inverse correlation between downregulation of hsa-miR-130a-3p and upregulation of hsa_circRNA_102610 in CD patients was observed (r = -0.290, P = 0.024) by Pearson correlation analysis. Moreover, overexpression of hsa_circRNA_102610 promoted SMAD4 and CyclinD1 protein expression validated by western-blotting. Furthermore, over-expression of hsa_circRNA_102610 promoted TGF-ß1 induced EMT in HIECs and NCM460 cells via targeting of hsa-miR-130a-3p, with increased expression of Vimentin and N-cadherin and decreased expression of E-cadherin. CONCLUSION: Hsa_circRNA_102610 upregulation in CD patients could promote the proliferation and EMT of intestinal epithelial cells via sponging of hsa-miR-130a-3p.