Yong Jiang, He Jun, Ning Feng
Department of Urology, Hunan Children's Hospital, the Paediatric Academy of University of South China Changsha, Hunan, PR China.
Histol Histopathol. 2023 May;38(5):559-570. doi: 10.14670/HH-18-539. Epub 2022 Oct 26.
Circular RNA (circRNA) has been shown to play an essential role in cancer progression, including nephroblastoma. Hsa_circ_0093741 was discovered to be highly expressed in nephroblastoma. However, its function and mechanism in nephroblastoma development are still vague.
The expression levels of hsa_circ_0093741, miR-562 and FRS2 (Fibroblast Growth Factor Receptor Substrate 2) were detected using western blotting and quantitative real-time polymerase chain reaction. Functional experiments were performed by using cell counting kit-8, colony formation, 5-ethynyl-2'-deoxyuridine (EdU), transwell, scratch assays in vitro and animal experiments in vivo. The interaction analysis was conducted using dual-luciferase reporter assay and RIP assay.
Hsa_circ_0093741 was highly expressed in nephroblastoma tissues and cells. Functionally, hsa_circ_0093741 silencing significantly suppressed the growth, invasion, and migration of nephroblastoma cells in vitro. MiR-562 was decreased in nephroblastoma, and was validated to be a target of hsa_circ_0093741. Inhibition of miR-562 reversed the anticancer functions of hsa_circ_0093741 silencing on nephroblastoma cells. FRS2 expression was increased in nephroblastoma and served as a target of miR-562, moreover, FRS2 overexpression attenuated the inhibitory functions of miR-562 on the nephroblastoma cell malignant phenotypes mentioned above. Pre-clinically, lentivirus-mediated hsa_circ_0093741 silencing also impeded nephroblastoma tumor growth and metastasis in vivo.
Knockdown of hsa_circ_0093741 suppresses nephroblastoma cell growth, migration and invasion by regulating the miR-562/FRS2 axis, suggesting the potential involvement of hsa_circ_0093741 in nephroblastoma progression.
环状RNA（circRNA）已被证明在包括肾母细胞瘤在内的癌症进展中起重要作用。已发现hsa_circ_0093741在肾母细胞瘤中高表达。然而，其在肾母细胞瘤发生发展中的功能和机制仍不清楚。
采用蛋白质免疫印迹法和定量实时聚合酶链反应检测hsa_circ_0093741、miR-562和FRS2（成纤维细胞生长因子受体底物2）的表达水平。通过细胞计数试剂盒-8、集落形成、5-乙炔基-2'-脱氧尿苷（EdU）、transwell、体外划痕试验和体内动物实验进行功能实验。使用双荧光素酶报告基因检测和RNA免疫沉淀实验进行相互作用分析。
hsa_circ_0093741在肾母细胞瘤组织和细胞中高表达。在功能上，hsa_circ_0093741沉默显著抑制肾母细胞瘤细胞在体外的生长、侵袭和迁移。miR-562在肾母细胞瘤中表达降低，并被证实是hsa_circ_0093741的靶标。抑制miR-562可逆转hsa_circ_0093741沉默对肾母细胞瘤细胞的抗癌作用。FRS2在肾母细胞瘤中表达增加，是miR-562的靶标，此外，FRS2过表达减弱了miR-562对上述肾母细胞瘤细胞恶性表型的抑制作用。临床前研究中，慢病毒介导的hsa_circ_0093741沉默也可抑制肾母细胞瘤在体内的肿瘤生长和转移。
敲低hsa_circ_0093741通过调节miR-562/FRS2轴抑制肾母细胞瘤细胞的生长、迁移和侵袭，提示hsa_circ_0093741可能参与肾母细胞瘤的进展。
