Department of Pediatrics, Jining First People's Hospital, Jining, China.
Eur Rev Med Pharmacol Sci. 2020 Feb;24(3):1088-1097. doi: 10.26355/eurrev_202002_20159.
The aim of this study was to investigate the expression characteristics of lncRNA CRNDE in Wilms' tumor and to further investigate whether it could promote the development of Wilms' tumor via regulating microRNA-424.
Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was performed to examine the expression level of CRNDE in tumor tissues and para-cancerous tissues of patients with Wilms' tumor. Meanwhile, the expression of CRNDE in Wilms' tumor cell lines was analyzed as well. CRNDE overexpression and knockdown models were constructed using lentivirus transfection in HFWT and 17-94 cell lines, respectively. Subsequently, cell counting kit-8 (CCK-8), cell colony formation, and transwell assays were performed to explore the influence of CRNDE on the biological functions of Wilms' tumor cells. Furthermore, luciferase reporter gene assay and cell reversal experiment were applied to explore the interplay between CRNDE and microRNA-424.
RT-qPCR results revealed that the expression level of lncRNA CRNDE in tumor tissues of patients with Wilms' tumor was remarkably higher than that of adjacent normal tissues. Also, the difference was statistically significant (p<0.05). Compared with patients with low expression of CRNDE, the risk of lymph node metastasis was significantly higher in patients with high CRNDE expression (p<0.05). Similarly, compared with control group, the proliferation and metastasis abilities of cells in CRNDE knockdown group were remarkably down-regulated (p<0.05). However, opposite results were observed in CRNDE overexpression group. In addition, our results demonstrated that microRNA-424 expression was negatively correlated with CRNDE expression in Wilms' tumor tissues. Luciferase reporter gene assay indicated that CRNDE could be targeted by microRNA-424 through specific a binding site, further regulating the malignant progression of Wilms' tumor.
CRNDE was highly expressed in Wilms' tumor tissue and cell lines. The expression of CRNDE was correlated with the incidence rate of lymph node metastasis in patients with Wilms' tumor. In addition, CRNDE might accelerate the progression of Wilms' tumor via modulating microRNA-424.
本研究旨在探讨 lncRNA CRNDE 在肾母细胞瘤中的表达特征，并进一步探讨其是否可通过调节 microRNA-424 促进肾母细胞瘤的发展。
采用实时定量聚合酶链反应（RT-qPCR）检测肾母细胞瘤患者肿瘤组织和癌旁组织中 CRNDE 的表达水平。同时分析肾母细胞瘤细胞系中 CRNDE 的表达情况。分别通过慢病毒转染构建 HFWT 和 17-94 细胞系的 CRNDE 过表达和敲低模型。随后，通过细胞计数试剂盒-8（CCK-8）、细胞集落形成和 Transwell 实验研究 CRNDE 对肾母细胞瘤细胞生物学功能的影响。此外，还应用荧光素酶报告基因实验和细胞逆转实验来探讨 CRNDE 与 microRNA-424 之间的相互作用。
RT-qPCR 结果显示，肾母细胞瘤患者肿瘤组织中 lncRNA CRNDE 的表达水平明显高于癌旁正常组织，差异具有统计学意义（p<0.05）。与 CRNDE 低表达患者相比，CRNDE 高表达患者发生淋巴结转移的风险明显更高（p<0.05）。同样，与对照组相比，CRNDE 敲低组细胞的增殖和转移能力明显下调（p<0.05）。然而，CRNDE 过表达组则观察到相反的结果。此外，我们的研究结果表明，在肾母细胞瘤组织中，microRNA-424 的表达与 CRNDE 的表达呈负相关。荧光素酶报告基因实验表明，CRNDE 可通过特定的结合位点被 microRNA-424 靶向，从而进一步调节肾母细胞瘤的恶性进展。
CRNDE 在肾母细胞瘤组织和细胞系中高表达。CRNDE 的表达与肾母细胞瘤患者的淋巴结转移发生率相关。此外，CRNDE 可能通过调节 microRNA-424 加速肾母细胞瘤的进展。
