癌相关成纤维细胞衍生的外泌体通过 OIP5-AS1/miR-142-5p/PD-L1 轴促进肺癌进展。

Cancer-associated fibroblasts-derived exosomes promote lung cancer progression by OIP5-AS1/ miR-142-5p/ PD-L1 axis.

机构信息

Department of Cardiothoracic Surgery, Affiliated Hospital of Nantong University, Nantong, 226000, Jiangsu, China.

Department of Cardiothoracic Surgery, The First People's Hospital of Suqian, Suqian, 223800, Jiangsu, China.

出版信息

Mol Immunol. 2021 Dec;140:47-58. doi: 10.1016/j.molimm.2021.10.002. Epub 2021 Oct 13.

DOI:10.1016/j.molimm.2021.10.002

PMID:34653794

Abstract

Cancer-associated fibroblasts (CAFs) are the most important stromal cells in the tumor microenvironment (TEM) and have been reported to regulate various cancer development. Exosomes are considered important elements involved in intercellular communication and TME regulation, while the potential function of CAFs in lung cancer immunosuppressive microenvironments remains unknown. CAFs-derived exosomes (CAFs-exo) and normal fibroblasts (NFs)-derived exosomes (NFs-exo) were isolated by ultra-centrifugation and characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and western blot analysis. A549 cells were co-cultured with peripheral blood mononuclear cells (PBMCs). Flow cytometry assay was performed to detect the killing role of PBMCs on A549 cells. Bioinformatics and luciferase reporter assays were used to analyze the relationship among microRNA (miRNA), long non-coding RNA (lncRNA) and target gene. BALB/c mice were used to construct the lung cancer model by subcutaneous injection. Programmed death ligand 1 (PD-L1) was up-regulated in lung cancer tissues and cells. PD-L1 also up-regulated in CAFs cell medium-mediated A549 cells. CAFs decreased PBMCs induced-cell apoptosis through increasing PD-L1 in A549 cells. Moreover, CAFs transferred exosomes to lung cancer cells to suppress the killing effect of PBMCs through up-regulating PD-L1. Using microarray assays, opa-interacting protein 5 antisense RNA 1 (OIP5-AS1) level was highly expressed in CAFs-exos. After treatment by CAFs-exos, miR-142-5p level was significantly down-regulated in A549 cells. OIP5-AS1 served as a sponge to target miR-142-5p and negatively regulated miR-142-5p expression in lung cancer cells. In addition, PD-L1 was a direct target of miR-142-5p. CAFs derived exosomal OIP5-AS1 reduced PBMCs induced-cell apoptosis and promoted tumor growth through decreasing miR-142-5p and up-regulating PD-L1. CAFs-derived exosomes suppressed the role of PBMCs induced-killing of lung cancer cells and promoted lung cancer progression by OIP5-AS1/ miR-142-5p/ PD-L1 axis, which provided a potential opportunity for diagnosis and treatment of lung cancer.

摘要

癌症相关成纤维细胞（CAFs）是肿瘤微环境（TEM）中最重要的基质细胞，已被报道可调节多种癌症的发展。外泌体被认为是细胞间通讯和 TME 调节的重要组成部分，而 CAFs 在肺癌免疫抑制微环境中的潜在功能尚不清楚。通过超速离心分离 CAFs 衍生的外泌体（CAFs-exo）和正常成纤维细胞（NFs）衍生的外泌体（NFs-exo），并通过透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和 Western blot 分析进行特征分析。将 A549 细胞与外周血单核细胞（PBMCs）共培养。通过流式细胞术检测 PBMCs 对 A549 细胞的杀伤作用。生物信息学和荧光素酶报告基因分析用于分析 microRNA（miRNA）、长非编码 RNA（lncRNA）和靶基因之间的关系。使用 BALB/c 小鼠通过皮下注射构建肺癌模型。程序性死亡配体 1（PD-L1）在肺癌组织和细胞中上调。CAFs 细胞培养基介导的 A549 细胞中 PD-L1 也上调。CAFs 通过增加 A549 细胞中的 PD-L1 来减少 PBMCs 诱导的细胞凋亡。此外，CAFs 通过外泌体将肺癌细胞转移到肺癌细胞中，通过上调 PD-L1 抑制 PBMCs 的杀伤作用。通过微阵列分析，OPA 相互作用蛋白 5 反义 RNA 1（OIP5-AS1）在 CAFs-exos 中高表达。用 CAFs-exos 处理后，A549 细胞中的 miR-142-5p 水平显著下调。OIP5-AS1 作为 miR-142-5p 的海绵，负调控肺癌细胞中 miR-142-5p 的表达。此外，PD-L1 是 miR-142-5p 的直接靶标。CAFs 衍生的外泌体 OIP5-AS1 通过降低 miR-142-5p 和上调 PD-L1，减少 PBMCs 诱导的细胞凋亡并促进肿瘤生长。CAFs 衍生的外泌体通过 OIP5-AS1/miR-142-5p/PD-L1 轴抑制 PBMCs 诱导的杀伤肺癌细胞的作用，促进肺癌进展，为肺癌的诊断和治疗提供了潜在的机会。