Liu Zheng, Wang Ronghai, Zhu Guangze
Department of Traditional Chinese Medicine and Acupuncture, Central Hospital of Baoji, Baoji City, Shaanxi Province, People's Republic of China.
Department of Urology, Linzi District People's Hospital, Zibo City, Shandong Province, People's Republic of China.
Cancer Manag Res. 2021 Jan 25;13:693-706. doi: 10.2147/CMAR.S282806. eCollection 2021.
Renal cell carcinoma (RCC) that originates from the proximal renal tubules is the most common cancer of the human kidney. Increasing circRNA/miRNA/mRNA networks have been found in RCC regulation. This study will explore the regulatory relation of circular RNA (circRNA) circ_0035483, microRNA-31-5p (miR-31-5p) and high mobility group A1 (HMGA1).
The levels of circ_0035483, miR-31-5p and HMGA1 were measured by real-time polymerase chain reaction (qRT-PCR) or Western blot. Cell proliferation was determined using Cell Counting Kit-8 (CCK-8) and colony formation assays. Cell migration and invasion were assessed by transwell assay. HMGA1 and epithelial-mesenchymal transition (EMT)-related protein levels were quantified using Western blot. Glycolytic metabolism was evaluated by glucose consumption and lactate production. The interaction between targets was confirmed via dual-luciferase reporter, RNA immunoprecipitation (RIP) and RNA pull-down assays. experiment was performed through the establishment of xenograft models in mice.
Circ_0035483 expression was upregulated in RCC tissues and cells. The inhibitory effects on RCC cell proliferation, migration, invasion, EMT and glycolysis were induced after circ_0035483 was downregulated. MiR-31-5p was identified as a target of circ_0035483 and miR-31-5p upregulation was related to the function of circ_0035483 knockdown in RCC cells. Additionally, miR-31-5p targeted HMGA1 and inhibited the malignant behaviors of RCC cells by negatively regulating HMGA1. Moreover, HMGA1 expression was regulated by circ_0035483 via targeting miR-31-5p. Circ_0035483 also affected tumor growth by relying on the miR-31-5p/HMGA1 axis.
These findings clarified that the tumor-promoting function of circ_0035483 in RCC was partly achieved by regulating the miR-31-5p/HMGA1 axis.
起源于近端肾小管的肾细胞癌（RCC）是人类肾脏最常见的癌症。在RCC调控中发现越来越多的环状RNA/微小RNA/信使核糖核酸网络。本研究将探讨环状RNA（circRNA）circ_0035483、微小RNA-31-5p（miR-31-5p）和高迁移率族蛋白A1（HMGA1）的调控关系。
采用实时聚合酶链反应（qRT-PCR）或蛋白质免疫印迹法检测circ_0035483、miR-31-5p和HMGA1的水平。使用细胞计数试剂盒-8（CCK-8）和集落形成试验测定细胞增殖。通过Transwell试验评估细胞迁移和侵袭。使用蛋白质免疫印迹法定量HMGA1和上皮-间质转化（EMT）相关蛋白水平。通过葡萄糖消耗和乳酸产生评估糖酵解代谢。通过双荧光素酶报告基因、RNA免疫沉淀（RIP）和RNA下拉试验确认靶标之间的相互作用。通过在小鼠中建立异种移植模型进行实验。
Circ_0035483在RCC组织和细胞中表达上调。下调circ_0035483后，可诱导对RCC细胞增殖、迁移、侵袭、EMT和糖酵解的抑制作用。MiR-31-5p被鉴定为circ_0035483的靶标，miR-31-5p上调与RCC细胞中circ_0035483敲低的功能相关。此外，miR-31-5p靶向HMGA1并通过负调节HMGA1抑制RCC细胞恶性行为。此外，HMGA1表达受circ_0035483通过靶向miR-31-5p调控。Circ_0035483还通过依赖miR-31-5p/HMGA1轴影响肿瘤生长。
这些发现阐明了circ_0035483在RCC中的促肿瘤功能部分是通过调节miR-31-5p/HMGA1轴实现的。
