Department of Pediatric Surgery, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, China.
Anticancer Drugs. 2022 Sep 1;33(8):720-730. doi: 10.1097/CAD.0000000000001320. Epub 2022 Aug 10.
Wilms' tumor (WT) is the most typical basic renal tumor in children and is associated with a high recurrence rate and improper diagnosis. Long noncoding RNAs (lncRNAs) play important roles in WT development. However, the impact of the OSTM1 antisense RNA 1 (OSTM1-AS1) lncRNA on WT remains largely unexplored. Differential expression of OSTM1-AS1, miR-514a-3p and maternal embryonic leucine zipper kinase (MELK) in mice with WT cells was assessed via quantitative reverse transcription-PCR and western blotting. Changes in the proliferation, migration and apoptosis of WT cells after OSTM1-AS1, miR-514a-3p or MELK knockdown were assessed using the cell counting kit-8, Transwell and caspase-3 activity assays, respectively. Additionally, the tumorigenicity of WT cells after OSTM1-AS1 knockdown in vivo was analyzed using a xenograft tumor assay. The association among OSTM1-AS1, MELK and miR-514a-3p was confirmed using the RNA binding protein immunoprecipitation and luciferase reporter assays. OSTM1-AS1 and MELK were upregulated in WT cells, whereas miR-514a-3p was downregulated. OSTM1-AS1 was mostly observed in the cytoplasm, and its knockout suppressed WT cell migration and proliferation in vitro , triggered apoptosis and attenuated tumor development in vivo . MiR-514a-3p was sponged by OSTM1-AS1, and miR-514a-3p interference counteracted the tumoricidal effect of OSTM1-AS1 knockdown. MiR-514a-3p reduced WT progression by downregulating the expression of MELK, which is the target gene of miR-514a-3p. lncRNA OSTM1-AS1 acts as an oncogenic factor in WT by releasing MELK through sponging miR-514a-3p and could be a useful target for WT diagnosis and therapy.
威尔姆斯瘤（WT）是儿童中最典型的基本肾肿瘤，与高复发率和不当诊断有关。长链非编码 RNA（lncRNA）在 WT 发育中发挥重要作用。然而，OSTM1 反义 RNA 1（OSTM1-AS1）lncRNA 对 WT 的影响在很大程度上仍未得到探索。通过定量逆转录 PCR 和 Western blot 评估 WT 细胞中 OSTM1-AS1、miR-514a-3p 和母源性胚胎亮氨酸拉链激酶（MELK）的差异表达。通过细胞计数试剂盒-8、Transwell 和 caspase-3 活性测定分别评估 OSTM1-AS1、miR-514a-3p 或 MELK 敲低后 WT 细胞的增殖、迁移和凋亡变化。此外，通过异种移植肿瘤测定分析 WT 细胞中 OSTM1-AS1 敲低后的体内致瘤性。通过 RNA 结合蛋白免疫沉淀和荧光素酶报告基因测定证实了 OSTM1-AS1、MELK 和 miR-514a-3p 之间的关联。在 WT 细胞中上调 OSTM1-AS1 和 MELK，而下调 miR-514a-3p。OSTM1-AS1 主要存在于细胞质中，其敲除抑制 WT 细胞体外迁移和增殖，触发凋亡并减弱体内肿瘤发生。miR-514a-3p 被 OSTM1-AS1 吸收，miR-514a-3p 干扰抵消了 OSTM1-AS1 敲低的抗肿瘤作用。miR-514a-3p 通过下调 MELK 的表达来减少 WT 的进展，MELK 是 miR-514a-3p 的靶基因。lncRNA OSTM1-AS1 通过海绵吸附 miR-514a-3p 释放 MELK，在 WT 中作为致癌因子发挥作用，可能成为 WT 诊断和治疗的有用靶点。
