Department of Ophthalmology, Shenzhen Eye Hospital, Shenzhen Eye Institute, Jinan University, No. 18 Zetian Road, Futian District, Shenzhen, 518040, Guangdong Province, China.
Department of Ophthalmology, The Second People's Hospital of Shenzhen, Shenzhen, Guangdong Province, China.
Int Ophthalmol. 2022 Feb;42(2):509-523. doi: 10.1007/s10792-021-02067-7. Epub 2021 Oct 11.
It has been gradually recognized that circular RNAs (circRNAs) are important modulators in multiple malignancies. Here, we analyzed the function of circ_0075804 and explored its associated mechanism in regulating retinoblastoma (RB) progression.
Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot assay were utilized to measure RNA and protein expression, respectively. Cell proliferation was analyzed by Cell counting kit-8 (CCK8) assay and 5-Ethynyl-2'-deoxyuridine (EdU) assay. Cell apoptosis was assessed by flow cytometry. Cell migration and invasion abilities were analyzed by wound healing assay and transwell invasion assay. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were applied to verify intermolecular target relations. Xenograft tumor model was used to analyze the role of circ_0075804 in tumor growth in vivo.
Circ_0075804 expression was markedly up-regulated in RB tissues and cell lines. Circ_0075804 knockdown restrained the proliferation, migration and invasion whereas promoted the apoptosis of RB cells. Circ_0075804 acted as a molecular sponge for microRNA-138-5p (miR-138-5p), and circ_0075804 silencing-induced effects were partly reversed by miR-138-5p knockdown in RB cells. MiR-138-5p interacted with the 3' untranslated region (3'UTR) of paternally expressed 10 (PEG10). Circ_0075804 positively regulated PEG10 level by sponging miR-138-5p in RB cells. PEG10 overexpression largely overturned miR-138-5p overexpression-mediated effects in RB cells. Circ_0075804 knockdown blocked xenograft tumor growth in vivo.
Circ_0075804 promoted RB progression via miR-138-5p-dependent regulation of PEG10, which provided new insight in RB therapy.
环状 RNA（circRNAs）已逐渐被认为是多种恶性肿瘤中的重要调节因子。在这里，我们分析了 circ_0075804 的功能，并探讨了其在调节视网膜母细胞瘤（RB）进展中的相关机制。
采用逆转录定量聚合酶链反应（RT-qPCR）和 Western blot 检测分别测量 RNA 和蛋白质表达。通过细胞计数试剂盒-8（CCK8）检测和 5-乙炔基-2'-脱氧尿苷（EdU）检测分析细胞增殖。通过流式细胞术评估细胞凋亡。通过划痕愈合试验和 Transwell 侵袭试验分析细胞迁移和侵袭能力。应用双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）试验验证分子间靶标关系。利用异种移植肿瘤模型分析 circ_0075804 在体内肿瘤生长中的作用。
circ_0075804 在 RB 组织和细胞系中表达明显上调。circ_0075804 敲低抑制 RB 细胞的增殖、迁移和侵袭，而促进细胞凋亡。circ_0075804 作为 microRNA-138-5p（miR-138-5p）的分子海绵，并且在 RB 细胞中，circ_0075804 沉默诱导的作用部分被 miR-138-5p 敲低逆转。miR-138-5p 与父系表达 10（PEG10）的 3'非翻译区（3'UTR）相互作用。circ_0075804 通过海绵 miR-138-5p 在 RB 细胞中正向调节 PEG10 水平。PEG10 的过表达在 RB 细胞中很大程度上推翻了 miR-138-5p 过表达介导的作用。circ_0075804 敲低阻断了体内异种移植肿瘤的生长。
circ_0075804 通过 miR-138-5p 依赖的 PEG10 调节促进 RB 进展，为 RB 治疗提供了新的见解。
