Department of Pediatrics, Second Hospital, Jilin University, Changchun, China.
The Key Laboratory of Pathobiology, Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Pediatr Dev Pathol. 2020 Mar-Apr;23(2):144-151. doi: 10.1177/1093526619873491. Epub 2019 Sep 9.
Wilms' tumor is the most common pediatric renal malignancy. MiRNAs are important regulators in multiple cancers including Wilms' tumor. In this study, we examined the role of miR-483-3p on proliferation, chemosensitivity, migration, and invasion of Wilms' tumor cells. The proliferation of Wilms' tumor cells was examined using WST-1 assay. The migration and invasion of Wilms' tumor cells were evaluated by transwell migration assay and matrigel invasion assay. The protein expression levels were detected by Western blot. The effect of miR-483-3p on doxorubicin-induced apoptosis in Wilms' tumor cells was evaluated by caspase-Glo3/7 assay. Forced expression of miR-483-3p promoted the proliferation, migration, and invasion in Wilms' tumor cells. Meanwhile, miR-483-3p decreased the sensitivity of Wilms' tumor cells after doxorubicin treatment. MiR-483-3p inhibited the doxorubicin-induced apoptosis in Wilms' tumor cells by the regulation of BAX and Bcl-2 expression. Furthermore, miR-483-3p regulated epithelial-mesenchymal transition by affecting the expression of E-cadherin, N-cadherin, snail, and vimentin in Wilms' tumor cells. Further studies showed that the expression levels of PTEN and p-AKT in Wilms' tumor cells were changed after aberrant expression of miR-483-3p by binding to 3'-UTR of PTEN. Our study suggests that miR-483-3p played important roles in proliferation and progression in Wilms' tumor cells and might serve as a potential prognostic biomarker and predict chemotherapy response in Wilms' tumor.
威尔姆斯瘤是最常见的小儿肾恶性肿瘤。miRNAs 是包括威尔姆斯瘤在内的多种癌症的重要调控因子。在这项研究中，我们研究了 miR-483-3p 对威尔姆斯瘤细胞增殖、化疗敏感性、迁移和侵袭的作用。使用 WST-1 法检测威尔姆斯瘤细胞的增殖。通过 Transwell 迁移实验和 Matrigel 侵袭实验评估威尔姆斯瘤细胞的迁移和侵袭。通过 Western blot 检测蛋白表达水平。通过 caspase-Glo3/7 assay 评估 miR-483-3p 对多柔比星诱导的威尔姆斯瘤细胞凋亡的影响。强制表达 miR-483-3p 促进了威尔姆斯瘤细胞的增殖、迁移和侵袭。同时，miR-483-3p 降低了多柔比星处理后威尔姆斯瘤细胞的敏感性。miR-483-3p 通过调节 BAX 和 Bcl-2 的表达抑制了多柔比星诱导的威尔姆斯瘤细胞凋亡。此外，miR-483-3p 通过影响 E-cadherin、N-cadherin、snail 和 vimentin 在威尔姆斯瘤细胞中的表达来调节上皮-间充质转化。进一步的研究表明，通过与 PTEN 的 3'-UTR 结合，miR-483-3p 异常表达后，威尔姆斯瘤细胞中 PTEN 和 p-AKT 的表达水平发生了变化。我们的研究表明，miR-483-3p 在威尔姆斯瘤细胞的增殖和进展中发挥重要作用，可能作为潜在的预后生物标志物和预测威尔姆斯瘤化疗反应的指标。
