Gong Jian, Song Zicheng, Pan Jiandong, Huang Xiangwei, Feng Xiaofen, Li Qian
Department of Clinical Laboratory, the Second Affiliated Hospital and Yuying Children's hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China.
State Key Laboratory of Ophthalmology, Optometry and Vision Science, Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
J Cancer. 2025 Apr 21;16(8):2434-2448. doi: 10.7150/jca.99599. eCollection 2025.
Retinoblastoma (RB) is a prevalent intraocular malignant tumor, posing a significant threat to human life and health. Although the involvement of circRNAs in various malignancies has been reported, their precise role in RB remains incompletely understood. High-throughput sequencing was utilized to construct differential expression profiles of circRNAs, followed by candidate gene screening using RT-qPCR. The circular structure of circSLC39A8 was confirmed through stability assays with RNase R and Act D. A research system for transiently silencing and overexpressing circSLC39A8 was established, with flow cytometry employed to assess cell cycle and apoptosis levels. Bioinformatics analyses, RT-qPCR, and western blot experiments were conducted to evaluate the expression of circSLC39A8, hsa-miR-11181-5p, and PIK3CA. RNA antisense purification experiments were performed to elucidate interactions among circSLC39A8, hsa-miR-11181-5p, and PIK3CA. Our findings revealed a significant upregulation of circSLC39A8 in RB. Functionally, circSLC39A8 was identified as promoting cellular proliferation and suppressing apoptosis , thereby facilitating RB progression. Mechanistically, circSLC39A8 indirectly augmented the expression levels of PIK3CA mRNA by acting as a competitive endogenous RNA for hsa-miR-11181-5p, consequently enhancing the stability of PIK3CA mRNA and ultimately fostering RB cell proliferation while inhibiting apoptosis.
视网膜母细胞瘤(RB)是一种常见的眼内恶性肿瘤,对人类生命和健康构成重大威胁。尽管已有报道称环状RNA(circRNA)参与了各种恶性肿瘤的发生发展,但其在RB中的具体作用仍未完全明确。本研究利用高通量测序构建circRNA差异表达谱,随后通过逆转录定量聚合酶链反应(RT-qPCR)筛选候选基因。通过核糖核酸酶R(RNase R)和放线菌素D(Act D)稳定性实验证实了circSLC39A8的环状结构。建立了circSLC39A8瞬时沉默和过表达的研究体系,采用流式细胞术评估细胞周期和凋亡水平。通过生物信息学分析、RT-qPCR和蛋白质免疫印迹实验评估circSLC39A8、hsa-miR-11181-5p和磷脂酰肌醇-3激酶催化亚基α(PIK3CA)的表达。进行RNA反义纯化实验以阐明circSLC39A8、hsa-miR-11181-5p和PIK3CA之间的相互作用。我们的研究结果显示,RB中circSLC39A8显著上调。在功能上,circSLC39A8被确定为促进细胞增殖和抑制细胞凋亡,从而推动RB进展。机制上,circSLC39A8作为hsa-miR-11181-5p的竞争性内源性RNA间接增加了PIK3CA mRNA的表达水平,进而增强了PIK3CA mRNA的稳定性,最终促进RB细胞增殖并抑制凋亡。