Hsa_circ_0046523 Mediates an Immunosuppressive Tumor Microenvironment by Regulating MiR-148a-3p/PD-L1 Axis in Pancreatic Cancer.
Hsa_circ_0046523 Mediates an Immunosuppressive Tumor Microenvironment by Regulating MiR-148a-3p/PD-L1 Axis in Pancreatic Cancer.
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Hsa_circ_0046523 通过调节胰腺癌中的 MiR-148a-3p/PD-L1 轴介导免疫抑制肿瘤微环境
DOI：
10.3389/fonc.2022.877376
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发表时间：
2022
影响因子：
4.7
通讯作者：
中科院分区：
文献类型：
--
作者：
环状RNA（CircRNA）是一类新型的非编码RNA，在肿瘤的发生发展中起着重要作用。然而，circRNA在调节胰腺癌（PC）免疫微环境中的功能和机制仍不清楚。CCK 8法和Transwell法检测hsa_circ_0046523表达对PC细胞增殖、迁移和侵袭能力的影响。采用流式细胞仪检测共培养后外周血单个核细胞（PBMC）中CD 4 + T细胞、CD 8 + T细胞和T淋巴细胞的比例，以及CD 8 + T细胞的凋亡、耗竭和功能。酶联免疫吸附试验（ELISA）检测细胞因子的表达水平。使用双荧光素酶报告基因测定hsa_circ_0046523、miR-148 a-3 p和PD-L1之间的相互作用。采用拯救实验和PD-L1阻断实验来研究hsa_circ_0046523是否通过PC中的miR-148 a-3 p/PD-L1发挥其生物学功能。此外，建立了免疫活性小鼠PC模型来证实这些发现。Hsa_circ_0046523在PC组织和细胞系中的表达显著上调。hsa_circ_0046523高表达与肿瘤分期及预后相关。Hsa_circ_0046523过表达促进PC细胞的增殖、迁移和侵袭。共培养实验证实，hsa_circ_0046523的强制表达可降低外周血单个核细胞（PBMC）中的CD 4+和CD 8 + T细胞的比例，以及增加TcB的比例。同时，hsa_circ_0046523过表达促进了CD 8 + T细胞的凋亡和耗竭，抑制了CD 8 + T细胞的功能，增加了PBMC中免疫抑制细胞因子IL-10和TGF-β的分泌，减少了免疫效应细胞因子IFN-γ和IL-2的分泌。hsa_circ_0046523通过与miR-148 a-3 p结合上调PC中PD-L1表达而发挥其生物学功能。此外，miR-148 a-3 p/PD-L1轴的这些免疫调节功能也在免疫活性小鼠PC模型中得到证实。我们的研究表明hsa_circ_0046523/miR-148 a-3 p/PD-L1调节轴介导PC免疫抑制微环境，这些分子有望成为重塑PC肿瘤免疫微环境的新靶点。
Circular RNAs (circRNAs) are a novel type of non-coding RNA, play an important role in the progression of tumors. However, the function and mechanism of circRNAs in regulating immune microenvironment of pancreatic cancer (PC) remain largely unclear. The effects of hsa_circ_0046523 expression on proliferation, migration and invasion of PC cells were analyzed by CCK8 and Transwell assays. Flow cytometry was used to detect the proportion of CD4+ T cells, CD8+ T cells and Tregs in peripheral blood mononuclear cells (PBMCs) after co-culture, and the apoptosis, depletion and function of CD8+ T cells. The expression levels of immunoregulatory cytokines were detected by enzyme linked immunosorbent assay (ELISA). The dual-luciferase reporter was performed to determine the interaction between hsa_circ_0046523, miR-148a-3p, and PD-L1. Rescue experiments and PD-L1 blocking experiments were employed to investigate whether hsa_circ_0046523 exerts its biological function by miR-148a-3p/PD-L1 in PC. Furthermore, an immunocompetent murine PC model was established to confirm these findings. Hsa_circ_0046523 expression was remarkably upregulated in PC tissues and cell lines. Moreover, high expression of hsa_circ_0046523 was correlated with advanced pathological stage and poorer prognosis. Hsa_circ_0046523 overexpression promoted the proliferation, migration and invasion of PC cells in vitro. Co-culture experiments confirmed that forced expression of hsa_circ_0046523 could decrease the proportion of CD4+ and CD8+ T cells, as well as increase the proportion of Tregs among peripheral blood mononuclear cells (PBMCs). Meanwhile, hsa_circ_0046523 overexpression promoted the apoptosis and exhaustion of CD8+ T cells, inhibited CD8+ T cell function, increased the secretion of immunosuppressive cytokines IL-10 and TGF-β, and decreased the secretion of immune effector cytokines IFN-γ and IL-2 among PBMCs. Mechanistically, hsa_circ_0046523 exerted its biological function by binding to miR-148a-3p to upregulate PD-L1 expression in PC. Moreover, these immune modulating functions of miR-148a-3p/PD-L1 axis were also confirmed in an immunocompetent murine PC model. Our study suggests that hsa_circ_0046523/miR-148a-3p/PD-L1 regulatory axis mediates PC immunosuppressive microenvironment and these molecules are expected to be new targets for remodeling tumor immune microenvironment of PC.
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影响因子：
3.9
作者：
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通讯作者：
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