Miao Xiaolin, Wang Zhen, Chen Bingyu, Chen Yiqi, Wang Xi, Jiang Luxi, Jiang Shanshan, Hao Ke, Zhang Wei
Outpatient Department, Eye Hospital of Wenzhou Medical University, China.
Research Center of Blood Transfusion Medicine, Education Ministry Key Laboratory of Laboratory Medicine, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou medical College, China.
Cell Mol Biol (Noisy-le-grand). 2018 May 15;64(6):42-47.
Retinoblastoma (RB) is a childhood intraocular tumor, affecting millions of patients worldwide. MicroRNA-140-5p (miR-140-5p) was demonstrated to be involved in the tumorigenesis of various human cancers; however, its role in RB remains undetermined. In this study, quantitative real-time PCR (qRT-PCR) and Western blot assays were used to determine the expression levels of miR-140-5p, cell migration-inducing protein (CEMIP), and cell adhesion molecule 3 (CADM3) in RB tissues and cell-lines. The proliferation ability was detected by cell-counting kit 8 (CCK-8), Edu staining, and colony formation assay. The cell cycle and migration and invasion abilities were measured by flow cytometry, wound-healing assay and Transwell assays, respectively. The correlation between miR-140-5p and CEMIP/CADM3 were then confirmed by immunofluorescence (IF) and dual-luciferase reporter assays. The results showed that miR-140-5p expression was significantly decreased; however, CEMIP and CADM3 expression was increased in RB tissues and cells. Overexpression of miR-140-5p inhibited proliferation, migration, and invasion of RB cells. We also found that miR-140-5p inhibited CEMIP and CADM3 expressions in RB cells. In addition, we demonstrated that miR-140-5p might negatively regulate the transcriptional activities of CEMIP and CADM3 by targeting their 3'-UTR. Therefore, we suggested that miR-140-5p could be a potential therapeutic target for the treatment of RB through CEMIP and CADM3.
视网膜母细胞瘤（RB）是一种儿童眼内肿瘤，全球数百万患者受其影响。MicroRNA-140-5p（miR-140-5p）已被证明参与多种人类癌症的肿瘤发生过程；然而，其在RB中的作用仍未明确。在本研究中，采用定量实时PCR（qRT-PCR）和蛋白质免疫印迹法检测miR-140-5p、细胞迁移诱导蛋白（CEMIP）和细胞黏附分子3（CADM3）在RB组织和细胞系中的表达水平。通过细胞计数试剂盒8（CCK-8）、Edu染色和集落形成试验检测增殖能力。分别通过流式细胞术、伤口愈合试验和Transwell试验测定细胞周期以及迁移和侵袭能力。然后通过免疫荧光（IF）和双荧光素酶报告基因试验证实miR-140-5p与CEMIP/CADM3之间的相关性。结果显示，miR-140-5p表达显著降低；然而，CEMIP和CADM3在RB组织和细胞中的表达增加。miR-140-5p过表达抑制RB细胞的增殖、迁移和侵袭。我们还发现miR-140-5p抑制RB细胞中CEMIP和CADM3的表达。此外，我们证明miR-140-5p可能通过靶向CEMIP和CADM3的3'-UTR负调控其转录活性。因此，我们认为miR-140-5p可能通过CEMIP和CADM3成为治疗RB的潜在治疗靶点。
