| Recent studies have indicated that miRNAs play a vital role in tumor development and usually regulated gene expression in the post transcription. In glioma, the research on the relationship between miRNAs and SALL4 and their multiple targets with same seed region haven not reported.To investigate the function of miR-103, miR-195 and miR-15 b, a series of experiments were performed. Overexpression of miR-103, miR-195 and miR-15 b suppressed cell proliferation by MTT assay. To verify the mechanism of suppression, we utilized flow cytometer by double staining of PI/Annexin V, and the results confirmed that the apoptosis of glioma cells were induced by miR-103, miR-195 and miR-15 b. Besides, the 3 miRNAs compete with each other.In addition, forced expression of miR-103, miR-195 and miR-15 b could inhibit the ability of migration and invasion by Transwell experiments. In different extent. Morover, co-expression of the 3 miRNAs indicated that there is a mutual competition between them.The analysis of miR-103, miR-195 and miR-15 b expression in glioma tissues, which is detected by real-time quantitative PCR, demonstrated that the expression level of the 3 mi RNAs were declined significantly in over 60% of glioma tissues. Comparing with the results of SALL4 expression of the same samples in our former project, we used SPSS software to analyze the correlations between SALL4 and the 3 candidate miRNAs. Our results indicated that SALL4 is nagetively correlated with the expression of miR-103, miR-195 and miR-15 b. The result gained by bioinformatics software showed that SALL4 is a probable target gene of both miR-103, miR-195 and miR-15 b, and they had a same “seed†portion. The results obtained by dual-luciferase reporter system assay, qRT-PCR and Western Blot evidenced that the expression of SALL4 protein would be competitively suppressed when miR-103, mi R-195 and miR-15 b was co-transfected, thus, it have proved that SALL4 is a direct target of the 3 miRNAs. |
PDF Full Text Request