Exosomal microRNAs in the DLK1-DIO3 imprinted region derived from cancer-associated fibroblasts promote progression of hepatocellular carcinoma by targeting hedgehog interacting protein.
Exosomal microRNAs in the DLK1-DIO3 imprinted region derived from cancer-associated fibroblasts promote progression of hepatocellular carcinoma by targeting hedgehog interacting protein.
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源自癌症相关成纤维细胞的 DLK1-DIO3 印迹区域中的外泌体 microRNA 通过靶向刺猬蛋白相互作用促进肝细胞癌的进展
DOI：
10.1186/s12876-022-02594-2
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发表时间：
2022-12-08
影响因子：
2.4
通讯作者：
Wang, Bei-Li
中科院分区：
文献类型：
--
作者：
Jin, An-Li;Ding, Lin;Yang, Wen-Jing;Liu, Te;Chen, Wei;Li, Tong;Zhang, Chun-Yan;Pan, Bai-Shen;Qiu, Shuang-Jian;Zhou, Jian;Fan, Jia;Guo, Wei;Yang, Xin-Rong;Wang, Bei-Li
关键词：
到2020年，肝细胞癌（HCC）是全球第六大最常诊断的癌症，也是癌症相关死亡的第三大原因。源自癌症相关成纤维细胞（CAFs-exo）的外泌体可以促进多种人类癌症的肿瘤进展。然而，控制CAFs-exo如何促进HCC进展的潜在调节机制仍然知之甚少。从HCC组织及相应的癌旁组织中分离出CAFs和癌旁成纤维细胞（paf），并进行体外培养。采用免疫荧光和免疫印迹（WB）方法对caf和paf进行表征。通过超离心分离外泌体，并通过透射电镜、纳米流式细胞术和WB测定对其进行表征。荧光显微镜下观察肝癌细胞外泌体的内化。细胞计数试剂盒-8 （CCK-8）检测细胞增殖情况。伤口愈合和transwell实验用于迁移和侵袭实验。采用RT-PCR法检测外泌体和HCC细胞中差异表达的microRNAs （miRNAs）。TargetScan数据库用于预测miRNA靶基因。使用TIMER、UALCAN、Kaplan-Meier绘图仪和LinkedOmics数据库进行刺猬相互作用蛋白（HHIP）表达分析、预后分析和HHIP相关共表达基因富集分析。CAFs-exo被HCC细胞内化。CAFs-exo有助于HCC细胞的侵袭性表型，而抑制外泌体分泌可逆转这些作用。在机制上，与PAFs-exo相比，与CAFs-exo共培养的HCC细胞中DLK1-DIO3印迹区域的mirna （miR-329-3p, miR-380-3p, miR-410-5p, miR-431-5p）增加。HHIP是一种可能的miR-431-5p靶基因，在HCC细胞中表达显著下调。肿瘤组织中HHIP低表达水平可预测HCC患者预后不良。hhip相关共表达基因主要与细胞粘附分子相关。CAFs-exo可以通过将DLK1-DIO3印迹区域的mirna传递到HCC细胞，从而抑制HHIP的表达，从而促进HCC的进展。hip是HCC的潜在预后生物标志物。在线版本包含补充材料，可在10.1186/s12876-022-02594-2获得。
Hepatocellular carcinoma (HCC) is the sixth most commonly diagnosed cancer and third leading cause of cancer-related death worldwide in 2020. Exosomes derived from cancer-associated fibroblasts (CAFs-exo) can promote tumor progression in various human cancers. However, the underlying regulatory mechanism controlling how CAFs-exo can promote HCC progression remains poorly understood. CAFs and para-cancer fibroblasts (PAFs) were isolated from HCC tissues and corresponding para-cancer tissues, then were cultured in vitro. CAFs and PAFs were characterized by immunofluorescence and western blot (WB) assays. Exosomes were isolated by ultracentrifugation, and characterized by transmission electron microscopy, nanoflow cytometry, and WB assay. The internalization of exosomes by HCC cells was observed under a fluorescence microscope. Cell Counting Kit-8 (CCK-8) assay was used to evaluate cell proliferation. Wound healing and transwell assays were used for migration and invasion experiments. RT-PCR assay was used to examine differentially expressed microRNAs (miRNAs) in exosomes and HCC cells. The TargetScan database was used to predict miRNA target genes. Hedgehog interacting protein (HHIP) expression analysis, prognostic analysis, and enrichment analysis of HHIP-related co-expressed genes were performed using the TIMER, UALCAN, Kaplan–Meier plotter, and LinkedOmics databases. CAFs-exo were internalized by HCC cells. CAFs-exo contributed to the aggressive phenotype of HCC cells, while inhibiting exosome secretion reversed these effects. Mechanistically, miRNAs in the DLK1-DIO3 imprinted region (miR-329-3p, miR-380-3p, miR-410-5p, miR-431-5p) were increased in HCC cells co-cultured with CAFs-exo compared with PAFs-exo. Expression of HHIP, a possible miR-431-5p target gene, was significantly downregulated in HCC cells. Low HHIP expression level in tumor tissues could predict poor prognosis in HCC patients. HHIP-related co-expressed genes were mainly associated with cell adhesion molecules. CAFs-exo can promote HCC progression by delivering miRNAs in the DLK1-DIO3 imprinted region to HCC cells, subsequently inhibiting HHIP expression. HHIP is a potential prognostic biomarker in HCC. The online version contains supplementary material available at 10.1186/s12876-022-02594-2.
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DOI：
10.18632/aging.203283
发表时间：
2021-07-29
期刊：
Aging
影响因子：
--
作者：
Xu J;Lu W
通讯作者：
Lu W
影响因子：
4.7
作者：
Song Y;Tu J;Cheng Y;Zhou F;Liu P;Zhou S;Gu Y;Sun Y
通讯作者：
Sun Y
血浆外泌体hsa_circ_0070396对肝细胞癌的诊断价值
影响因子：
2.2
作者：
Lyu, Lihua;Yang, Wenjing;Guo, Wei
通讯作者：
Guo, Wei
影响因子：
3.5
作者：
Chang, Cui;Liu, Te;Chen, Juan
通讯作者：
Chen, Juan
影响因子：
11.2
作者：
Li T;Fan J;Wang B;Traugh N;Chen Q;Liu JS;Li B;Liu XS
通讯作者：
Liu XS