Zhao Long, Zhang Haoran, Wang Shuo, Zhou Yushi, Jiang Kewei, Wang Shan, Ye Yingjiang, Wang Bo, Shen Zhanlong
Department of Gastroenterological Surgery, Peking University People's Hospital, Beijing, China.
Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, China.
Mol Carcinog. 2025 Jun;64(6):1025-1038. doi: 10.1002/mc.23906. Epub 2025 Mar 26.
Accumulating evidence indicated that circular RNAs (circRNAs) directly sponge to microRNAs (miRNAs),(miRNAs), which in turn regulate the gene expression and affect the malignancy behavior at the posttranscriptional level. However, the expression levels, function, and mechanism of circ_0000231 in colorectal cancer (CRC) are largely unknown. The expression levels of circ_0000231, miR-140-3p, and Bcl-2 in 110 CRC tissues and matched normal colorectal tissues were detected by qRT-PCR method. circ_0000231 and Bcl-2 were suppressed by siRNA, and miR-140-3p was overexpressed by RNA mimics in CRC cell lines. The function-based experiments were conducted to detect the proliferation and migratory abilities in CRC cell lines. RNA pull-down, RNA immunoprecipitation (RIP), and dual-fluorescence reporter assay were conducted to verify the association among circ_0000231, miR-140-3p, and Bcl-2. Western blot analysis and mRFP-GFP-LC3 adenovirus were used to detect the autophagy level affected by circ_0000231, miR-140-3p, and Bcl-2 axis. Downregulated circ_0000231 significantly suppressed the proliferation and migratory abilities of CRC cells by suppressing autophagy and promoting G0/G1 phase arrest. Furthermore, RNA pull-down, RIP, and dual-fluorescence reporter assays confirmed that circ_0000231 regulates the expression of Bcl-2 by directly targeting miR-140-3p. More importantly, circ_0000231 promoted the levels of autophagy via the miR-140-3p/Bcl-2 axis in CRC. Our study demonstrated that circ_0000231, as a tumor promotor, enhances the level of autophagy by regulating Bcl-2 via targeting miR-140-3p. Moreover, circ_0000231 might serve as a diagnostic and prognostic indicator and a novel molecular target for CRC therapy.
越来越多的证据表明，环状RNA（circRNAs）直接吸附微小RNA（miRNAs），而miRNAs反过来在转录后水平调节基因表达并影响恶性行为。然而，circ_0000231在结直肠癌（CRC）中的表达水平、功能及机制仍 largely未知。采用qRT-PCR法检测110例CRC组织及配对的正常结直肠组织中circ_0000231、miR-140-3p和Bcl-2的表达水平。在CRC细胞系中，circ_0000231和Bcl-2被siRNA抑制，miR-140-3p被RNA模拟物过表达。进行基于功能实验检测CRC细胞系中的增殖和迁移能力。采用RNA下拉、RNA免疫沉淀（RIP）和双荧光报告基因检测法验证circ_0000231、miR-140-3p和Bcl-2之间的关联。采用蛋白质免疫印迹分析和mRFP-GFP-LC3腺病毒检测circ_0000231、miR-140-3p和Bcl-2轴影响的自噬水平。下调的circ_0000231通过抑制自噬和促进G0/G1期阻滞显著抑制CRC细胞的增殖和迁移能力。此外，RNA下拉、RIP和双荧光报告基因检测证实circ_0000231通过直接靶向miR-140-3p调节Bcl-2的表达。更重要的是，circ_系列0000231通过CRC中的miR-140-3p/Bcl-2轴促进自噬水平。我们的研究表明，circ_000系列000231作为一种肿瘤促进因子，通过靶向miR-140-3p调节Bcl-2来提高自噬水平。此外，circ_0000231可能作为CRC诊断和预后指标以及一种新的分子治疗靶点。
