Department of General Surgery, Shenzhen Children's Hospital, Yitian Road 7019, Shenzhen, 518000, Guangdong, China.
Mol Biotechnol. 2023 Dec;65(12):2030-2037. doi: 10.1007/s12033-023-00714-1. Epub 2023 Mar 14.
In order to examine new potential treatment options for the treatment of hepatoblastoma (HB), we identified the differential expression of five-candidate tumor suppressor miRNAs in HB and explored possible regulatory mechanisms of target miRNA molecule. By using real-time quantitative polymerase chain reaction (qPCR), we examined the expression of miRNAs in HB tissues and cells. The effect of has-miR-139-3p mimics on the invasion and migration ability was assessed by transwell assay and scratch-wound assay in HepG2 cells. Subsequently, we analyzed the target genes of miR-139-3p and their enrichment signaling pathways through bioinformatics. qPCR, Western-blot and dual-luciferase assays were further used to assess whether has-miR-139-3p targets Wnt5A. The results showed that hsa-miR-139-3p was significantly decreased in HB cells. Upregulation of hsa-miR-139-3p inhibited the invasive and migratory ability of HepG2. Bioinformatics analysis showed that hsa-miR-139-3p may target Wnt5A to regulate the WNT pathway, which was further confirmed by Western-blot and dual-luciferase assays. Overexpression of Wnt5A can reverse the miR-139-3p mimic-induced declines in the expression of WNT pathway-related proteins and restore the invasion and migration of HepG2. These data indicated that the hsa-miR-139-3p/Wnt5A axis inhibited HB metastasis, suggesting that miR-139-3p and Wnt5A may be potential targets for the treatment of HB.
为了研究新的潜在治疗方法治疗肝母细胞瘤(HB),我们鉴定了五种候选肿瘤抑制 miRNA 在 HB 中的差异表达,并探讨了靶 miRNA 分子的可能调控机制。通过实时定量聚合酶链反应(qPCR),我们检测了 HB 组织和细胞中 miRNA 的表达。通过 Transwell 测定和划痕试验评估了 has-miR-139-3p 模拟物对 HepG2 细胞侵袭和迁移能力的影响。随后,我们通过生物信息学分析了 miR-139-3p 的靶基因及其富集信号通路。qPCR、Western-blot 和双荧光素酶测定进一步用于评估 has-miR-139-3p 是否靶向 Wnt5A。结果表明,hsa-miR-139-3p 在 HB 细胞中显著降低。hsa-miR-139-3p 的上调抑制了 HepG2 的侵袭和迁移能力。生物信息学分析表明,hsa-miR-139-3p 可能靶向 Wnt5A 调节 WNT 通路,这进一步通过 Western-blot 和双荧光素酶测定得到证实。Wnt5A 的过表达可以逆转 miR-139-3p 模拟物诱导的 WNT 通路相关蛋白表达下降,并恢复 HepG2 的侵袭和迁移。这些数据表明 hsa-miR-139-3p/Wnt5A 轴抑制 HB 转移,提示 miR-139-3p 和 Wnt5A 可能是 HB 治疗的潜在靶点。