Dual targeting of PD-L1 and PD-L2 by PCED1B-AS1 via sponging hsa-miR-194-5p induces immunosuppression in hepatocellular carcinoma
Dual targeting of PD-L1 and PD-L2 by PCED1B-AS1 via sponging hsa-miR-194-5p induces immunosuppression in hepatocellular carcinoma
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DOI：
10.1007/s12072-020-10101-6
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发表时间：
2020-11-21
影响因子：
6.6
通讯作者：
Wu, Bin
中科院分区：
文献类型：
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作者：
Fan, Fei;Chen, Keji;Wu, Bin
PD-L1和PD-L2是PD-1配体（PD-Ls）。PD-Ls过表达与肝细胞癌（HCC）预后不良相关。然而，我们对PD-Ls的表达是如何调控的知之甚少。在这里，我们研究了lncRNA-microRNA网络在HCC中参与PD-Ls的调节。方法检测45对HCC标本中PD-Ls、PCED1B-AS1和hsa-miR-194-5p的表达。通过microRNA下拉和体外结合实验检测PCED1B-AS1与hsa-miR-194-5p的相互作用。研究了PCED1B-AS1敲低和过表达对HCC细胞系中hsa-miR-194-5p和PD-Ls表达的影响。观察HCC细胞系与人T细胞共培养的免疫抑制作用。从肝癌细胞中分离外泌体并研究其对受体细胞的影响。通过体外和体内实验评估肿瘤行为。结果肝癌组织中PD-L1表达与PD-L2表达高度相关。PCED1B-AS1和hsa-miR-194-5p在HCC中表达上调。在HCC中，PCED1B-AS1与PD-Ls呈正相关，与hsa-miR-194-5p呈负相关。这些相关性通过TCGA-LIHC数据集进行了交叉验证。PCED1B-AS1与hsa-mir-194-5p相互作用，抑制PD-Ls的表达。PCED1B-AS1通过海绵处理hsa-mir-194-5p增强PD-Ls的表达。pced1b - as1诱导pd - ls介导的共培养T细胞免疫抑制。HCC细胞释放含有外泌体的PCED1B-AS1，外泌体PCED1B-AS1增强受体HCC细胞中PD-Ls的表达，同时抑制受体T细胞和巨噬细胞。血外泌体PCED1B-AS1与HCC PD-Ls表达相关。最后，PCED1B-AS1促进异种移植裸鼠细胞增殖、集落形成和体内肿瘤形成，抑制细胞凋亡。结论PCED1B-AS1通过海绵化hsa-miR-194-5p，增强PD-Ls的表达和功能，诱导HCC的免疫抑制。图形抽象
Background PD-L1 and PD-L2 are PD-1 ligands (PD-Ls). PD-Ls over-expression is associated with poor prognosis in hepatocellular carcinoma (HCC). However, little is known about how PD-Ls expression is regulated. Here, we investigated the involvement of lncRNA-microRNA network in the regulation of PD-Ls in HCC. Methods The expression of PD-Ls, PCED1B-AS1 and hsa-miR-194-5p was measured in 45 pairs of HCC samples. The interaction between PCED1B-AS1 and hsa-miR-194-5p was measured by microRNA pull down and in vitro binding assay. The effects of PCED1B-AS1 knockdown and over-expression on hsa-miR-194-5p and PD-Ls expression were investigated in HCC cell lines. Immunosuppression was evaluated in co-culture of HCC cell line and human T cells. Exosomes were isolated from HCC cells and their effects on receipt cells were investigated. Tumor behaviors were evaluated by in vitro and in vivo assays. Results PD-L1 expression was highly correlated with PD-L2 expression in HCC. PCED1B-AS1 and hsa-miR-194-5p expression was up-regulated in HCC. PCED1B-AS1 was positively correlated with PD-Ls but negatively correlated hsa-miR-194-5p in HCC. These correlations were cross-validated by TCGA-LIHC dataset. PCED1B-AS1 interacted with hsa-mir-194-5p which inhibited PD-Ls expression. PCED1B-AS1 enhanced the expression of PD-Ls via sponging hsa-mir-194-5p. PCED1B-AS1-induced PD-Ls-mediated immunosuppression in co-cultured T cells. HCC cells released PCED1B-AS1 containing exosomes and the exosomal PCED1B-AS1 enhanced PD-Ls expression in receipt HCC cells while inhibited receipt T cells and macrophages. Blood exosomal PCED1B-AS1 was correlated with HCC PD-Ls expression. Finally, PCED1B-AS1 promoted cell proliferation, colony formation and in vivo tumor formation in xenografted nude mice while inhibited apoptosis. Conclusions PCED1B-AS1 enhances the expression and function of PD-Ls via sponging hsa-miR-194-5p to induce immunosuppression in HCC. Graphic abstract