Deng Zhenwei, Li Xiyao, Wang Huaiming, Geng Yongyong, Cai Yongchang, Tang Yuxin, Wang Yijun, Yu Xueqiao, Li Libo, Li Ruiping
Department of General Surgery, Dongguan People's Hospital, Southern Medical University, Dongguan, China.
Department of General Surgery, The First Hospital of China Medical University, Shenyang, China.
Front Genet. 2020 May 8;11:357. doi: 10.3389/fgene.2020.00357. eCollection 2020.
This study was aimed to evaluate the potential function of circ-0001946 in the progression of colorectal cancer (CRC) and the related regulatory mechanism. First, the expression levels of circRNA_0001946 and microRNA-135a-5p (miR-135a-5p) in normal and CRC tissues were measured by quantitative real-time polymerase chain reaction (RT-qPCR). In addition, cell proliferation was assessed by the Cell Counting Kit-8 (CCK-8) assay, cell migration and invasion were evaluated by Transwell assays, and the cell cycle patterns were determined by flow cytometry. The relationship between the expression levels of circ_0001946 and miR-135a-5p was determined by dual-luciferase reporter assays. Our data showed that the expression of circ_0001946 was upregulated in CRC tissues, which was negatively correlated with tumor size, histologic grade, lymphatic metastasis, and TMN stage, and patients with circ_0001946 overexpression were more likely to have a poor prognosis. In addition, experiments showed that silencing circ_0001946 inhibited the epithelial-mesenchymal transition (EMT) pathway and markedly suppressed CRC cell growth, migration, and invasion. Furthermore, we discovered that the transfection of miR-135a-5p mimics could reverse the antitumor effects of circRNA_0001946 downregulation. To summarize, this study revealed that circRNA_0001946 might act as a tumor promoter by activating the miR-135a-5p/EMT axis and may be a promising treatment target for CRC.
本研究旨在评估circ-0001946在结直肠癌（CRC）进展中的潜在作用及其相关调控机制。首先，通过定量实时聚合酶链反应（RT-qPCR）检测正常组织和CRC组织中circRNA_0001946和微小RNA-135a-5p（miR-135a-5p）的表达水平。此外，采用细胞计数试剂盒-8（CCK-8）法评估细胞增殖，通过Transwell实验评估细胞迁移和侵袭能力，并通过流式细胞术确定细胞周期模式。通过双荧光素酶报告基因实验确定circ_0001946与miR-135a-5p表达水平之间的关系。我们的数据显示，circ_0001946在CRC组织中表达上调，这与肿瘤大小、组织学分级、淋巴转移和TMN分期呈负相关，且circ_0001946过表达的患者预后更差。此外，实验表明，沉默circ_0001946可抑制上皮-间质转化（EMT）途径，并显著抑制CRC细胞的生长、迁移和侵袭。此外，我们发现转染miR-135a-5p模拟物可逆转circRNA_0001946下调的抗肿瘤作用。综上所述，本研究表明circRNA_0001946可能通过激活miR-135a-5p/EMT轴发挥肿瘤促进作用，可能是CRC的一个有前景的治疗靶点。
