Children's Medical Center of The First Hospital of Changsha, Changsha, Hunan, People's Republic of China.
Children's Medical Center of The Second Xiangya Hospital of Central South University, Changsha, Hunan, People's Republic of China.
Technol Cancer Res Treat. 2021 Jan-Dec;20:15330338211036523. doi: 10.1177/15330338211036523.
Wilm's tumor is a common renal malignancy in childhood with unsatisfactory prognosis. microRNA-215-5p (miR-215-5p) has been reported as a tumor-suppressive miRNA in different types of human cancers, but rarely in the Wilm's tumor. In light of this, we tried to investigate the regulatory role and underlying mechanism of miR-215-5p in the Wilm's tumor.
After sample collection and cell culture, the expression of miR-215-5p and CT10 Regulator of Kinase (CRK) was detected. Then rhabdoid tumor cell lines (formerly classified as Wilms' tumor cell lines), G401 and WT-CLS1 cells were transfected with pcDNA3.1, pcDNA3.1-CRK, sh-NC, sh-CRK, agomir NC, miR-215-5p agomir, antagomir NC or miR-215-5p antagomir to explore the function of miR-215-5p and CRK in the Wilm's tumor cell proliferation and migration. Moreover, the relationship between miR-215-5p and CRK was analyzed by dual luciferase reporter gene assay.
Lowly-expressed miR-215-5p and highly-expressed CRK were observed in the Wilm's tumor tissues and cells. Transfection of pcDNA3.1-CRK or miR-215-5p antagomir could promote G401 and WT-CLS1 cell proliferation and enhance migration ability, while transfection of sh-CRK or miR-215-5p agomir led to opposite results. Additionally, miR-215-5p may bind to CRK. Moreover, transfection of pcDNA3.1-CRK in G401 and WT-CLS1 cells could partially reverse the inhibitory effect of miR-215-5p agomir on the proliferation and migration of Wilm's tumor cells.
Our study highlighted that miR-215-5p could suppress the proliferation and migration of Wilm's tumor cells by regulating the expression of CRK, providing new ideas for molecular targeted therapy for Wilm's tumor.
肾母细胞瘤是儿童期常见的肾恶性肿瘤，预后不佳。microRNA-215-5p（miR-215-5p）已被报道为多种人类癌症中的肿瘤抑制 miRNA，但在肾母细胞瘤中很少见。有鉴于此，我们试图研究 miR-215-5p 在肾母细胞瘤中的调节作用及其潜在机制。
采集样本并进行细胞培养后，检测 miR-215-5p 和 CT10 激酶调节蛋白（CRK）的表达。然后用 pcDNA3.1、pcDNA3.1-CRK、sh-NC、sh-CRK、agomir NC、miR-215-5p agomir、antagomir NC 或 miR-215-5p antagomir 转染横纹肌瘤细胞系（以前归类为肾母细胞瘤细胞系）G401 和 WT-CLS1 细胞，以探讨 miR-215-5p 和 CRK 在肾母细胞瘤细胞增殖和迁移中的作用。此外，通过双荧光素酶报告基因检测分析 miR-215-5p 和 CRK 之间的关系。
在肾母细胞瘤组织和细胞中观察到低表达的 miR-215-5p 和高表达的 CRK。pcDNA3.1-CRK 或 miR-215-5p antagomir 的转染可促进 G401 和 WT-CLS1 细胞的增殖并增强迁移能力，而 sh-CRK 或 miR-215-5p agomir 的转染则导致相反的结果。此外，miR-215-5p 可能与 CRK 结合。此外，pcDNA3.1-CRK 在 G401 和 WT-CLS1 细胞中的转染可部分逆转 miR-215-5p agomir 对肾母细胞瘤细胞增殖和迁移的抑制作用。
本研究强调，miR-215-5p 可通过调节 CRK 的表达抑制肾母细胞瘤细胞的增殖和迁移，为肾母细胞瘤的分子靶向治疗提供了新的思路。
