Toxicology Graduate Program, Faculty of Science, Mahidol University, Bangkok, Thailand; Excellent Center for Drug Discovery (ECDD), Mahidol University, Bangkok, Thailand.
Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Life Sci. 2018 Oct 1;210:65-75. doi: 10.1016/j.lfs.2018.08.058. Epub 2018 Aug 27.
Cholangiocarcinoma (CCA) is a malignant tumor of bile duct epithelial cells. The prognosis of CCA is poor due to lack of effective therapeutic targets and detection at an advanced stage. Exosomes are secreted nano-sized vesicles and contribute to the malignancy of several cancers via transferring their miRNAs between cells. Thus, exosomal miRNAs may serve as new therapeutic targets and potential biomarkers for CCA.
Exosomes were isolated from three different CCA cell lines and normal human cholangiocyte cells, followed by miRNA profiling analysis. Potential role of dysregulated miRNA was investigated by knockdown experiment.
We found that 38 and 460 miRNAs in CCA exosomes were significantly up- and down-regulated, respectively. Of these differentially expressed miRNAs, the hsa-miR-205-5p and miR-200 family members were markedly up-regulated for 600-1500 folds, whereas the miR-199 family members and their clustered miRNA, hsa-miR-214-3p, were down-regulated for 1000-2000 folds. The expression patterns of these representative exosomal miRNAs were similar to those observed in all types of CCA cells. The target genes of the top ten most up- and down-regulated miRNAs are significantly associated with well-characterized cancer-related pathways. Consistently, knockdown of the most up-regulated miRNA, miR-205-5p, reduced KKU-M213 cell invasion and migration.
We have demonstrated the distinct miRNA signatures in exosomes released from CCA cells, compared to normal human cholangiocyte cells. These exosomal miRNAs may have the potential to be novel therapeutic targets and biomarkers for CCA.
胆管癌（CCA）是一种胆管上皮细胞的恶性肿瘤。由于缺乏有效的治疗靶点和在晚期检测，CCA 的预后较差。外泌体是一种分泌的纳米大小囊泡，通过在细胞间转移其 miRNA 而促进几种癌症的恶性程度。因此，外泌体 miRNA 可能作为 CCA 的新治疗靶点和潜在的生物标志物。
从三种不同的 CCA 细胞系和正常人类胆管细胞中分离出外泌体，然后进行 miRNA 谱分析。通过敲低实验研究失调 miRNA 的潜在作用。
我们发现 CCA 外泌体中有 38 个和 460 个 miRNA 分别显著上调和下调。在这些差异表达的 miRNA 中，hsa-miR-205-5p 和 miR-200 家族成员上调了 600-1500 倍，而 miR-199 家族成员及其聚类 miRNA hsa-miR-214-3p 下调了 1000-2000 倍。这些代表性外泌体 miRNA 的表达模式与所有类型的 CCA 细胞中观察到的相似。上调和下调倍数最大的前 10 个 miRNA 的靶基因与已明确的癌症相关途径显著相关。一致地，下调最上调的 miRNA，miR-205-5p，减少了 KKU-M213 细胞的侵袭和迁移。
我们已经证明了 CCA 细胞释放的外泌体中独特的 miRNA 特征，与正常人类胆管细胞相比。这些外泌体 miRNA 可能具有作为 CCA 的新型治疗靶点和生物标志物的潜力。
