环状 RNA 0008287 通过损害 microRNA-548c-3p 依赖的 CLIC1 抑制作用促进胃癌细胞的免疫逃逸。
Circ_0008287 promotes immune escape of gastric cancer cells through impairing microRNA-548c-3p-dependent inhibition of CLIC1.
机构信息
Department of Gastrointestinal Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, P.R. China; Guangxi Key Laboratory of Enhanced Recovery after Surgery for Gastrointestinal Cancer, Nanning 530021, P.R. China.
Department of General Surgery, the Second Affiliated Hospital of Guangxi Medical University, Nanning 530007, P.R. China.
出版信息
Int Immunopharmacol. 2022 Oct;111:108918. doi: 10.1016/j.intimp.2022.108918. Epub 2022 Jul 26.
BACKGROUND
Analyses in silico suggested the upregulation of a circular RNA (circRNA), circ_0008287, in gastric cancer and possible interactions among microRNA (miR)-548c-3p, circ_0008287, and intracellular chloride channel protein 1 (CLIC1). This study aims to testify whether circ_0008287 can affect the immune escape of gastric cancer cells by regulating miR-548c-3p and CLIC1.
METHODS
RT-qPCR was performed to determine the expression pattern of circ_0008287 in gastric cancer cells. Gain- and loss-of function assays were then performed to assess the effects of circ_0008287 on malignant phenotypes of cancer cells. Interactions among circ_0008287, miR-548c-3p and CLIC1 were verified by dual luciferase reporter gene, RIP and FISH assays. Effects of CLIC1 on IFN-γ secretion and apoptosis in CD8 + T cells were evaluated by flow cytometry following co-culture of CD8 + T cells with cancer cells overexpressing/silencing CLIC1. A gastric cancer mouse model was further developed for in vivo investigation on effects of circ_0008287 on tumorigenesis and tumor metastasis.
RESULTS
circ_0008287, an upregulated circRNA in gastric cancer cells, augmented the viability as well as invasive and migratory potentials of gastric cancer cells. By competitively binding to miR-548c-3, circ_0008287 increased the expression of CLIC1, which impaired the function of CD8 + T cells and promoted their apoptosis. After downregulation of circ_0008287, in vivo tumorigenesis and metastasis were suppressed.
CONCLUSION
Hence, this study suggests the promotive role of circ_0008287 in gastric cancer progression and immune escape and further elucidates the underlying circ_0008287/miR-548c-3p/CLIC1 regulatory axis.
背景
基于计算机的分析表明，胃癌中环状 RNA（circRNA）circ_0008287 的表达上调，并且 microRNA（miR）-548c-3p、circ_0008287 和细胞内氯离子通道蛋白 1（CLIC1）之间可能存在相互作用。本研究旨在通过调节 miR-548c-3p 和 CLIC1 来验证 circ_0008287 是否可以影响胃癌细胞的免疫逃逸。
方法
通过 RT-qPCR 测定胃癌细胞中 circ_0008287 的表达模式。然后进行增益和缺失功能测定，以评估 circ_0008287 对癌细胞恶性表型的影响。通过双荧光素酶报告基因、RIP 和 FISH 测定验证 circ_0008287、miR-548c-3p 和 CLIC1 之间的相互作用。通过共培养过表达/沉默 CLIC1 的胃癌细胞与 CD8+T 细胞，通过流式细胞术评估 CLIC1 对 CD8+T 细胞 IFN-γ 分泌和凋亡的影响。进一步建立胃癌小鼠模型，以研究 circ_0008287 对肿瘤发生和转移的体内影响。
结果
在胃癌细胞中上调的 circ_0008287 增加了胃癌细胞的活力以及侵袭和迁移能力。通过竞争性结合 miR-548c-3，circ_0008287 增加了 CLIC1 的表达，从而损害了 CD8+T 细胞的功能并促进其凋亡。下调 circ_0008287 后，体内肿瘤发生和转移受到抑制。
结论
因此，本研究表明 circ_0008287 在胃癌进展和免疫逃逸中起促进作用，并进一步阐明了潜在的 circ_0008287/miR-548c-3p/CLIC1 调节轴。
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