Department of General Surgery, Jingmen No.1 People's Hospital, Jingmen, 448000, Hubei, China.
Department of Anorectal Surgery, Jingmen No.1 People's Hospital, No.167, Xiangshan Avenue, Dadao District, Jingmen, 448000, Hubei, China.
World J Surg Oncol. 2021 Feb 17;19(1):51. doi: 10.1186/s12957-021-02164-y.
Circular RNAs (circRNAs) are a class of endogenous single-strand RNA transcripts with crucial regulation in human cancers. The objective of this study is to investigate the role of circ_0082182 in CRC and its specific functional mechanism.
The quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure the levels of circ_0082182, microRNA-411 (miR-411) and microRNA-1205 (miR-1205). Cell proliferation was detected by Cell counting Kit-8 (CCK-8) and colony formation assays. Flow cytometry was used for determining cell cycle and cell apoptosis. Cell apoptosis was also assessed by caspase3 and caspase9 activities. Cell migration and invasion were examined using scratch assay and transwell assay. The interaction between circ_0082182 and miRNA was validated by the dual-luciferase reporter and biotinylated RNA pull-down assays. Wnt/β-catenin pathway and epithelial-mesenchymal transition (EMT)-associated proteins were quantified by Western blot. Xenograft model was established for the research of circ_0082182 in vivo.
Circ_0082182 was upregulated in CRC and could predict the poor prognosis of CRC patients. Functionally, circ_0082182 promoted CRC cell proliferation, cell cycle progression, and metastasis while inhibited apoptosis. Subsequently, circ_0082182 was shown to act as the sponges of miR-411 and miR-1205. MiR-411 and miR-1205 were identified as tumor inhibitors in CRC. Furthermore, circ_0082182 promoted the CRC progression via sponging miR-411 and miR-1205. Moreover, circ_0082182 facilitated the Wnt/β-catenin pathway and EMT process by targeting miR-411 and miR-1205. In vivo, circ_0082182 accelerated the CRC tumorigenesis and EMT process by activating the Wnt/β-catenin pathway by downregulating the expression of miR-411 or miR-1205.
This study showed that circ_0082182 functioned as an oncogene in the developing process of CRC by sponging miR-411 or miR-1205 to activate the Wnt/β-catenin pathway. Circ_0082182 might be a molecular target in the diagnosis and treatment of CRC.
环状 RNA（circRNAs）是一类具有重要调控作用的内源性单链 RNA 转录本，在人类癌症中。本研究的目的是探讨 circ_0082182 在 CRC 中的作用及其特定的功能机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 circ_0082182、microRNA-411（miR-411）和 microRNA-1205（miR-1205）的水平。细胞增殖采用细胞计数试剂盒-8（CCK-8）和集落形成实验检测。采用流式细胞术检测细胞周期和细胞凋亡。通过 caspase3 和 caspase9 活性评估细胞凋亡。通过划痕实验和 Transwell 实验检测细胞迁移和侵袭。通过双荧光素酶报告和生物素化 RNA 下拉实验验证 circ_0082182 与 miRNA 之间的相互作用。采用 Western blot 检测 Wnt/β-catenin 通路和上皮间质转化（EMT）相关蛋白的表达。建立裸鼠移植瘤模型研究 circ_0082182 在体内的作用。
circ_0082182 在 CRC 中上调，可预测 CRC 患者的不良预后。功能上，circ_0082182 促进 CRC 细胞增殖、细胞周期进程和转移，同时抑制细胞凋亡。随后，circ_0082182 被证明可以作为 miR-411 和 miR-1205 的海绵。miR-411 和 miR-1205 被鉴定为 CRC 中的肿瘤抑制因子。此外，circ_0082182 通过海绵吸附 miR-411 和 miR-1205 促进 CRC 进展。此外，circ_0082182 通过靶向 miR-411 和 miR-1205 促进 Wnt/β-catenin 通路和 EMT 过程。体内实验结果表明，circ_0082182 通过下调 miR-411 或 miR-1205 的表达，激活 Wnt/β-catenin 通路，加速 CRC 肿瘤发生和 EMT 过程。
本研究表明，circ_0082182 通过海绵吸附 miR-411 或 miR-1205 激活 Wnt/β-catenin 通路，在 CRC 的发生发展过程中发挥致癌基因的作用。circ_0082182 可能成为 CRC 诊断和治疗的分子靶点。
