Department of General Surgery, the First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, P.R. China.
Department of Developmental Biology, Jinzhou Medical University, Jinzhou, Liaoning, P.R. China.
PLoS One. 2021 Apr 26;16(4):e0250603. doi: 10.1371/journal.pone.0250603. eCollection 2021.
The pathogenesis of GC involves the complex networking of multiple signaling pathways; however, the detailed mechanisms of tumorigenesis of GC remains largely unknown. Therefore, it is necessary to explore novel diagnostic/prognostic biomarkers for GC. In this study, the levels of hsa_circRNA_100269 in gastric cancer (GC) samples and cells were examined, and its effects on the biological functions of GC cells were elucidated. The levels of hsa_circRNA_100269 in specimens/cell lines were examined using RT-qPCR. Cell models with hsa_circRNA_100269 overexpression or knockdown were generated using lentiviral vectors. Cell viability was determined by MTT assay; cell migratory/invasive activity was evaluated using wound healing/Transwell assay. Cell cycle arrest and apoptosis were assessed by flow cytometry; expression of associated markers involved in cell apoptosis, EMT and the PI3K/Akt signaling were determined by RT-qPCR/immunoblotting. In vivo study was also performed using hsa_circRNA_100269 knockout mice. Our findings revealed downregulation of hsa_circRNA_100269 in GC tissues compared to non-cancerous control. Additionally, the levels of PI3K were remarkably elevated in GC tissues, where hsa_circRNA_100269 and PI3K was negatively correlated. Moreover, the expression of hsa_circRNA_100269 was associated with histology grade and occurrence of metastasis in GC patients. In addition, hsa_circRNA_100269 was downregulated in GC cells compared to normal gastric epithelial cells. Overexpressed hsa_circRNA_100269 notably inhibited the proliferation, migration, invasion and EMT of GC cells, whereas cell cycle arrest at G0/G1 phase was promoted and cell apoptosis was enhanced. Moreover, the PI3K/Akt signaling was involved in hsa_circRNA_100269-regulated GC cell proliferation, migration, invasion, EMT and apoptosis. Knockdown of hsa_circRNA_100269 also remarkably induced tumor growth in mouse model. In summary, our findings indicated that the levels of hsa_circRNA_100269 were reduced in GC. Furthermore, hsa_circRNA_100269 could suppress the development of GC by inactivating the PI3K/Akt pathway. More importantly, hsa_circRNA_100269/PI3K/Akt axis may be a novel therapeutic candidate for GC treatment.
胃癌的发病机制涉及多种信号通路的复杂网络；然而，胃癌的肿瘤发生机制在很大程度上仍然未知。因此，有必要探索新的胃癌诊断/预后生物标志物。在这项研究中，检测了胃癌（GC）样本和细胞中 hsa_circRNA_100269 的水平，并阐明了其对 GC 细胞生物学功能的影响。使用 RT-qPCR 检测标本/细胞系中 hsa_circRNA_100269 的水平。使用慢病毒载体生成 hsa_circRNA_100269 过表达或敲低的细胞模型。通过 MTT 测定法确定细胞活力；通过划痕愈合/Transwell 测定法评估细胞迁移/侵袭活性。通过流式细胞术评估细胞周期停滞和细胞凋亡；通过 RT-qPCR/免疫印迹法测定与细胞凋亡、EMT 和 PI3K/Akt 信号相关的标志物的表达。还使用 hsa_circRNA_100269 敲除小鼠进行了体内研究。我们的研究结果显示，与非癌对照相比，GC 组织中 hsa_circRNA_100269 的表达下调。此外，GC 组织中 PI3K 的水平显著升高，hsa_circRNA_100269 与 PI3K 呈负相关。此外，hsa_circRNA_100269 的表达与 GC 患者的组织学分级和转移的发生有关。此外，与正常胃上皮细胞相比，GC 细胞中 hsa_circRNA_100269 的表达下调。过表达 hsa_circRNA_100269 显著抑制 GC 细胞的增殖、迁移、侵袭和 EMT，而促进细胞周期停滞在 G0/G1 期并增强细胞凋亡。此外，PI3K/Akt 信号通路参与了 hsa_circRNA_100269 调节的 GC 细胞增殖、迁移、侵袭、EMT 和凋亡。hsa_circRNA_100269 的敲低也显著诱导了小鼠模型中的肿瘤生长。总之，我们的研究结果表明，GC 中 hsa_circRNA_100269 的水平降低。此外，hsa_circRNA_100269 通过失活 PI3K/Akt 通路抑制 GC 的发展。更重要的是，hsa_circRNA_100269/PI3K/Akt 轴可能是治疗 GC 的新的治疗候选物。
