1 宜昌市第一人民医院，三峡大学人民医院肾内科，湖北 宜昌 443000)
目的：探究环状RNA(circRNA)0000218(circ_0000218)/miR-139-3p/SRY盒转录因子2(SOX2)分子轴在调节肾细胞癌(renal cell carcinoma，RCC)细胞增殖和转移的作用及其机制。方法：采用实时定量聚合酶链反应(qRT-PCR)检测肾癌组织和细胞中circ_0000218和miR-139-3p的表达水平，以及肾癌细胞中二者的调控关系，并对43例肾癌患者组织中circ_0000218和miR-139-3p的表达水平进行相关性分析。采用CCK-8和Transwell实验分别检测过表达circ_0000218或miR-139-3p模拟物在细胞中增殖和迁移侵袭的作用。采用StarBase数据库预测miR-139-3p在肾癌样本中的表达与其生存预后信息。采用蛋白质印迹法检测肾癌细胞中circ_0000218或miR-139-3p对SOX2表达水平的影响。最后采用CircInteractome和TargetScan预测，双荧光素酶报告基因实验验证circ_0000218与miR-139-3p，miR-139-3p和SOX2之间的靶向关系。结果：Circ_0000218在肾癌患者癌组织以及细胞系中均显著上调，同时其高表达水平和T分期级别增加，局部淋巴结转移和远处转移显著相关。过表达circ_0000218促进肾癌细胞增殖和转移。而miR-139-3p在肾癌组织和细胞中低表达，且StarBase数据库预测其低表达与肾癌患者生存期呈正相关。circ_0000218与miR-139-3p表达呈负相关，潜在的机制表明circ_0000218通过调节miR-139-3p/SOX2轴促进肾癌增殖和转移。结论：在RCC中，circ_0000218通过调节miR-139-3p/SOX2轴促进RCC细胞的增殖和转移，并有可能作为诊断性生物标志物和治疗靶点。
Circ_0000218 promoting the progression of renal cell carcinoma by regulating the molecular axis of miR-139-3p/SOX2
CorrespondingAuthor: CHEN Chang
This work was supported by the Major Teaching Research Project of China Three Gorges University, China (J2017056).
Objective: To explore the mechanism of circRNA (circ_0000218)/miR-139-3p/SRY-box transcription factor 2 (SOX2) axis in mediating the proliferation and metastasis of renal cell carcinoma cells. Methods: qRT-PCR was used to detect the expression of circ_0000218 and miR-139-3p in renal cell carcinoma tissues and cells, as well as the regulatory relationship between circ_0000218 and miR-139-3p in renal cancer cells. Subsequently, CCK-8 and Transwell experiments were used to detect the role of over expressed circ_0000218 or miR-139-3p mimics in cell proliferation, migration and invasion. The expression of miR-139-3p in RCC samples and its survival and prognosis information were predicted by StarBase database. Furthermore, Western blot was used to detect the effect of circ_0000218 or miR-139-3p on the expression of SOX2 in RCC cells. Finally, we use CircInteractome and TargetScan to predict, the double luciferase reporter gene experiment was verified the target relationship between circ_0000218 and miR-139-3p, miR-139-3p and SOX2. Results: Circ_0000218 was significantly up-regulated in renal cell carcinoma tissues and cell lines, and its high expression level was significantly correlated with T stage increased, local lymph node metastasis and distant metastasis. Overexpression of circ_0000218 promotes the proliferation and metastasis of RCC cells. The expression of miR-139-3p was low in renal cancer tissues and cells, and StarBase database predicted that its low expression was positively correlated with survival time of renal cancer patients. There is a negative correlation between circ_0000218 and miR-139-3p expression. The underlying mechanism suggests that circ_0000218 can promote the proliferation and metastasis of RCC by regulating miR-139-3p/SOX2 axis. Conclusion: In renal cell carcinoma, the circ_0000218 promotes the proliferation and metastasis of RCC cells by targeting on miR-139-3p/SOX2 axis, and may be used as a diagnostic bio-marker and therapeutic target.
renal cell carcinoma; circ_0000218; miR-139-3p; SRY-box transcription factor 2