Yan Guigang, Su Yi, Ma Zhao, Yu Lianzhi, Chen Ning
Department of Ophthalmology, the Affiliated Yantai Yuhuangding Hospital of Qingdao University.
Department of Radiation Oncology, the Affiliated Yantai Yuhuangding Hospital of Qingdao University.
Cell Struct Funct. 2019;44(1):51-60. doi: 10.1247/csf.18033.
Retinoblastoma (RB) is the most common intraocular malignancy in childhood, and the prognosis in the advanced RB is poor. It is urgent to find novel therapeutic targets. Long noncoding RNAs (lncRNAs) have critical functions in cancer progression, and lncRNA LINC00202 is found associated with poor prognosis in RB. However, the functions of LINC00202 in RB remain unclear. We employed qRT-PCR and immunoblot to detect the expression levels of mRNAs and proteins, respectively. Cell proliferation was determined by CCK-8 assay and colony formation assay. Transwell assays were applied to evaluate the cell abilities of migration and invasion. Luciferase reporter assay was applied to examine RNA stability, and RNA pulldown assays were used to detect interaction between lncRNA and microRNA (miRNA). LINC00202 expression in RB tissues is higher than that in the paired adjacent normal tissues, which has correlation with poor prognosis in RB. RB cell proliferation, migration and invasion were weakened by LINC00202 depletion, but enhanced by LINC00202 overexpression. MiR-3619-5p was identified to directly bind and mediate LINC00202-promoted RB progression, meanwhile, miR-3619-5p directly regulated expression of an oncongene, RIN1. Moreover, RIN1 knockdown completely blocked miR-3619-5p-enhanced RB progression. In summary, high LINC00202 levels are correlated with poor prognosis in RB, and it promotes RB progression by sponging miR-3619-5p and therefore up-regulating RIN1 expression.Key words: LINC00202, miR-3619-5p, retinoblastoma, progression, RIN1.
视网膜母细胞瘤(RB)是儿童期最常见的眼内恶性肿瘤,晚期RB的预后较差。迫切需要找到新的治疗靶点。长链非编码RNA(lncRNA)在癌症进展中具有关键作用,并且发现lncRNA LINC00202与RB的不良预后相关。然而,LINC00202在RB中的功能仍不清楚。我们分别采用qRT-PCR和免疫印迹法检测mRNA和蛋白质的表达水平。通过CCK-8法和集落形成试验测定细胞增殖。应用Transwell试验评估细胞的迁移和侵袭能力。采用荧光素酶报告基因试验检测RNA稳定性,并使用RNA下拉试验检测lncRNA与微小RNA(miRNA)之间的相互作用。RB组织中LINC00202的表达高于配对的相邻正常组织,这与RB的不良预后相关。LINC00202的缺失会削弱RB细胞的增殖、迁移和侵袭,但LINC00202的过表达则会增强这些能力。已确定miR-3619-5p直接结合并介导LINC00202促进的RB进展,同时,miR-3619-5p直接调节癌基因RIN1的表达。此外,RIN1的敲低完全阻断了miR-3619-5p增强的RB进展。总之,LINC00202的高表达水平与RB的不良预后相关,它通过吸附miR-3619-5p从而上调RIN1的表达来促进RB进展。关键词:LINC00202;miR-3619-5p;视网膜母细胞瘤;进展;RIN1