关键词: 视网膜母细胞瘤, miR-130b, 上调, PTEN, 促癌作用

Abstract:

Objective To detect the function and molecular mechanism of miR-130b in retinoblastoma (RB). Design Experimental study. Participants RB tissues and cells. Methods The expression of miR-130b in 40 pairs of RB tissues and adjacent normal tissues was detected by using quantitative RT-PCR (qRT-PCR). The MTT assay and Western blot analysis were performed to examine the proliferation and apoptosis abilities of RB cells. The target gene of miR-130b in RB cells was predicted with bioinformatics and verified with dual luciferase reporter assay and Western blot analysis. Main Outcome Measures The percentage of cell growth, the percentage of cell apoptosis and the activities of luciferase. Results The expression of miR-130b in RB tissues was significantly higher than matched adjacent normal tissues (P=0.023). miR-130b expression was also increased in HXO-Rb44 (P=0.008) and Y79 (P=0.012) cells compared with retinal microvascular endothelial cell ACBRI-181. Knock-down of miR-130b expression inhibited HXO-Rb44 (P=0.004) and Y79 (P=0.001) cells proliferation, and induced HXO-Rb44 (P=0.011) and Y79 (P=0.027) cells apoptosis. PTEN (Phosphatase and tensin homologue deleted on chromosome 10) was a target gene of miR-130b. The dual luciferase reporter assay and Western blot analysis showed that miR-130b could bind the mRNA 3’-UTR region of PTEN, inhibited translation of PTEN, and resulted in decreasing the protein expression of PTEN in (P=0.038) and Y79 (P=0.025) cells. Conclusions  miR-130b was up-regulated in RB tissues. miR-130b promoted RB cell growth and inhibited cell apoptosis targeting PTEN, suggesting miR-215 may serve as a novel biomarker in treatment of RB. (Ophthalmol CHN, 2017, 26: 276-281)

Key words: RB, miR-130b, up-regulation, PTEN, oncogene