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SKP2 受 circ_ODC1/miR-422a 轴的正向调节,促进视网膜母细胞瘤的增殖。

SKP2, positively regulated by circ_ODC1/miR-422a axis, promotes the proliferation of retinoblastoma.

机构信息

Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.

出版信息

J Cell Biochem. 2020 Jan;121(1):322-331. doi: 10.1002/jcb.29177. Epub 2019 Jul 11.

Abstract

Retinoblastoma (RB) is the most common intraocular malignancy in infants and children. S-phase kinase-associated protein 2 (SKP2) has been unmasked as an oncogene in a great many of carcinomas. The biologic function and the detailed molecular mechanism of SKP2 in RB need to be better understood. In this study, real-time quantitative polymerase chain reaction and Western blot showed the ectopic expression of SKP2 in RB tissues and cell lines. Loss of function assays showed the attenuated cell proliferation in RB as a result of SKP2 knockdown. In addition, bioinformatics analysis predicted the interaction between SKP2 and miR-422a. Luciferase reporter assay and Pearson's correlation analysis validated the negative correlation between miR-422a and SKP2. MiR-422a overexpression led to a decline of SKP2 expression and cell growth in RB. The binding capacity between miR-422a and circ_ODC1 was also predicted by bioinformatics analysis. Pearson's correlation analysis and luciferase reporter assay confirmed that circ_ODC1 is negatively correlated with miR-422a. Silencing circ_ODC1 resulted in a rise in miR-422a expression and RB cell growth. Moreover, reduced cell growth was restored by SKP2 overexpression. In a word, SKP2, induced by circ_ODC1 and miR-422a, promotes RB proliferation. Our new findings in this research might expedite the discovery of novel prognostic markers and therapeutic targets of RB.

摘要

视网膜母细胞瘤(RB)是婴儿和儿童中最常见的眼内恶性肿瘤。S 期激酶相关蛋白 2(SKP2)已被揭示为许多癌中的癌基因。SKP2 在 RB 中的生物学功能和详细的分子机制需要进一步了解。在这项研究中,实时定量聚合酶链反应和 Western blot 显示 SKP2 在 RB 组织和细胞系中的异位表达。功能丧失测定显示 SKP2 敲低导致 RB 细胞增殖减弱。此外,生物信息学分析预测了 SKP2 和 miR-422a 之间的相互作用。荧光素酶报告基因检测和 Pearson 相关分析验证了 miR-422a 和 SKP2 之间的负相关。miR-422a 的过表达导致 RB 中 SKP2 表达和细胞生长下降。生物信息学分析还预测了 miR-422a 和 circ_ODC1 之间的结合能力。Pearson 相关分析和荧光素酶报告基因检测证实 circ_ODC1 与 miR-422a 呈负相关。沉默 circ_ODC1 导致 miR-422a 表达和 RB 细胞生长增加。此外,通过过表达 SKP2 恢复了细胞生长的减少。总之,由 circ_ODC1 和 miR-422a 诱导的 SKP2 促进 RB 增殖。我们在这项研究中的新发现可能会加速 RB 新型预后标志物和治疗靶点的发现。

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