Zheng Wei, Liang Hong, Zhang Chao
Department of Gastrointestinal Surgery, Henan Provincial People's Hospital, Zhengzhou University People's Hospital, No. 7 Weiwu Road, Jinshui District, Zhengzhou, 450003, Henan, P.R. China.
Biochem Genet. 2025 Jun 26. doi: 10.1007/s10528-025-11162-8.
Colorectal cancer (CRC), as a common malignant tumor of gastrointestinal tract, has become one of the important diseases threatening human health. Circ_0001461 has been proven to be closely associated with the occurrence and development of various cancers. This study explored the function and possible mechanism of circ_0001461 in CRC. The RNA expression level was detected using qRT-PCR. The viability, apoptosis or invasion ability of CRC cells were explored by CCK-8, flow cytometry or Transwell experiment, respectively. The protein expression level in cells was analyzed by Western blot (WB). IFN-γ, IL-2 or TNF-α levels were investigated via ELISA experiment. The targeting relationship between circ_0001461, miR-532-3p and AMOTL2 was confirmed via dual-luciferase reporter experiment, RNA pull-down experiment and RIP assay. The protein expression in tissues was explored by IHC staining. Tumor volumes, body weights, tumor weights and apoptosis levels were detected in xenograft mouse model. Circ_0001461 was greatly expressed in CRC samples and was associated with poor prognosis. Silencing circ_0001461 reduced cell proliferation, increased cell apoptosis rate, decreased invasion ability and inhibited immune escape in vitro. Additionally, silencing circ_0001461 prominently shortened the tumor volume and tumor weight in vivo. Circ_0001461 targeted miR-532-3p to regulate AMOTL2 expression. AMOTL2 was the downstream targets of miR-532-3p. Downregulation of miR-532-3p or overexpression of AMOTL2 abolished the effect of silencing circ_0001461. Collectively, our research demonstrated that the circ_0001461 played a certain regulatory role in growth, movement and immune escape of colon cancer via miR-532-3p/AMOTL2 axis.
结直肠癌（CRC）作为一种常见的胃肠道恶性肿瘤，已成为威胁人类健康的重要疾病之一。Circ_0001461已被证明与多种癌症的发生和发展密切相关。本研究探讨了Circ_0001461在结直肠癌中的作用及可能机制。采用qRT-PCR检测RNA表达水平。分别通过CCK-8、流式细胞术或Transwell实验探讨结直肠癌细胞的活力、凋亡或侵袭能力。通过蛋白质免疫印迹法（WB）分析细胞中的蛋白质表达水平。通过ELISA实验检测IFN-γ、IL-2或TNF-α水平。通过双荧光素酶报告基因实验、RNA下拉实验和RIP分析证实Circ_0001461、miR-532-3p和AMOTL2之间的靶向关系。通过免疫组化染色探索组织中的蛋白质表达。在异种移植小鼠模型中检测肿瘤体积、体重、肿瘤重量和凋亡水平。Circ_0001461在结直肠癌样本中高表达，且与预后不良相关。沉默Circ_0001461可降低体外细胞增殖，增加细胞凋亡率，降低侵袭能力并抑制免疫逃逸。此外，沉默Circ_0001461可显著缩短体内肿瘤体积和肿瘤重量。Circ_0001461靶向miR-532-3p以调节AMOTL2表达。AMOTL2是miR-532-3p的下游靶点。下调miR-532-3p或过表达AMOTL2可消除沉默Circ_0001461的作用。总的来说，我们的研究表明Circ_0001461通过miR-532-3p/AMOTL2轴在结肠癌的生长、迁移和免疫逃逸中发挥一定的调节作用。
