MicroRNA‑506 regulates apoptosis in retinoblastoma cells by targeting sirtuin 1.

BACKGROUND

MicroRNAs have been reported to participate in the initiation and progression of retinoblastoma (RB), most common malignancy in children. The refractory mechanisms of chemoresistance and the toxicity of chemotherapies hindered the treatment especially on young children. Novel RB therapies are urgently required. MiR-506 is probed to be associated with the tumorigenesis of various cancers whilst the role of this miR in RB remains unclear.

METHODS

Here, the impact of miR-506 on RB cell survival in vitro and tumorigenesis in vivo was examined. MiR-506 expression was examined in human RB samples and cell lines as compared with healthy tissues and non-RB cells. EdU staining and colony formation assay were performed to determine the effect of miR-506 on RB cell growth. TdT-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry analysis were applied to detect the apoptotic cell number after miR-506 was downregulated in RB cells. Furthermore, dual-luciferase reporter assay was utilized to confirm the direct interaction between miR-506 and SIRT1 gene.

RESULTS

MiR-506 expression was upregulated in 20 human RB samples from patients as well as in human RB cell lines, WERI-Rb1 and Y79, as compared to that in healthy tissues and non-RB cells. In contrast, the expression of sirtuin 1 (SIRT1), known as NAD-dependent deacetylase, was downregulated in RB samples and cell lines. Aberrant reduced miR-506 expression impaired survival and proliferation of WERI-Rb1 and Y79 cells. The depletion of miR-506 expression promoted apoptosis of the two RB cell lines. The results of bioinformatics analysis and dual-luciferase assay exhibited that miR-506 targeted the 3'-untranslated region of SIRT1 on silencing purpose. The SIRT1 silencing lessened the miR-506 inhibition on RB cell proliferation and undermined apoptosis.

CONCLUSION

The results provided an insight into the role of miR-506 during RB development and offered potential pharmaceutical strategy for RB diagnosis.