Zhang Weiwei, Yang Qian, Qian Dongchen, Zhao Keli, Tang Chenxue, Ju Shaoqing
Department of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong, Jiangsu, P. R. China.
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong, Jiangsu, P. R. China.
Gastroenterol Rep (Oxf). 2024 Feb 28;12:goae008. doi: 10.1093/gastro/goae008. eCollection 2024.
Autophagy death of cancer cells is detrimental to apoptosis induced by therapeutic drugs, which promotes tumor progression to a certain extent. Increasing reports have demonstrated the regulatory role of circular RNAs (circRNAs) in autophagy. Here, we aimed to determine the role of hsa_circ_0009109 in autophagy in gastric cancer (GC).
The effects of hsa_circ_0009109 on autophagy were examined using quantitative real-time polymerase chain reaction (qPCR), transmission electron microscopy, Western blot, and immunofluorescence. The mechanism of hsa_circ_0009109 regulating the miR-544a-3p/bcl-2 axis was analysed using fluorescence hybridization, dual-luciferase reporter, and rescue experiments.
Functional testing indicated that hsa_circ_0009109 was significantly down-expressed in GC tissues and cell lines. A reduction in cytoplasmic-derived hsa_circ_0009109 could promote GC progression by accelerating cell proliferation, enhancing migration and invasion, inhibiting apoptosis, and accelerating the cell cycle progression. Besides, hsa_circ_0009109 was found to exert the effect of an autophagy inhibitor such as 3-Methyladenine (3-MA), which was manifested by the weakening of the immunofluorescence of LC3B and the reduction in autophagy-related proteins after overexpression of hsa_circ_0009109, while increased autophagosomes were observed after interference with hsa_circ_0009109. Subsequently, the crosstalk between hsa_circ_0009109 and miR-544a-3p/bcl-2 was verified using dual-luciferase reporter assay. The autophagy status was altered under the regulation of the hsa_circ_0009109-targeted miR-544a-3p/bcl-2 axis.
The hsa_circ_0009109 mediated a novel autophagy regulatory network through targeting the miR-544a-3p/bcl-2 axis, which may shed new light on the exploration of therapeutic targets for the clinical treatment of GC.
癌细胞的自噬性死亡不利于治疗药物诱导的细胞凋亡，在一定程度上促进肿瘤进展。越来越多的报道表明环状RNA（circRNA）在自噬中具有调控作用。在此，我们旨在确定hsa_circ_0009109在胃癌（GC）自噬中的作用。
使用定量实时聚合酶链反应（qPCR）、透射电子显微镜、蛋白质免疫印迹法和免疫荧光法检测hsa_circ_0009109对自噬的影响。使用荧光杂交、双荧光素酶报告基因和拯救实验分析hsa_circ_0009109调控miR-544a-3p/bcl-2轴的机制。
功能测试表明，hsa_circ_0009109在GC组织和细胞系中显著低表达。细胞质来源的hsa_circ_0009109减少可通过加速细胞增殖、增强迁移和侵袭、抑制细胞凋亡以及加速细胞周期进程来促进GC进展。此外，发现hsa_circ_0009109发挥了自噬抑制剂如3-甲基腺嘌呤（3-MA）的作用，表现为hsa_circ_0009109过表达后LC3B免疫荧光减弱和自噬相关蛋白减少，而干扰hsa_circ_0009109后观察到自噬体增加。随后，使用双荧光素酶报告基因检测验证了hsa_circ_0009109与miR-544a-3p/bcl-2之间的相互作用。在hsa_circ_0009109靶向的miR-544a-3p/bcl-2轴调控下，自噬状态发生改变。
hsa_circ_0009109通过靶向miR-544a-3p/bcl-2轴介导了一个新的自噬调控网络，这可能为GC临床治疗的治疗靶点探索提供新的思路。
