Gao Lili, Yang Xiaoping, Zhang Hao, Yu Minghua, Long Jianting, Yang Tao
Center for Medical Research and Innovation,
Department of General Surgery.
Onco Targets Ther. 2018 Oct 15;11:6981-6994. doi: 10.2147/OTT.S182225. eCollection 2018.
Cholangiocarcinoma (CCA) is epithelial cell malignancy with very poor prognosis. A lot of patients were diagnosed at advanced stage of CCA and no risk factors were identified. There are limited treatment options available for the management of CCA patients. It is urgent to develop effective targeted therapies for the treatment of CCA. miRNAs are small noncoding RNAs that negatively regulate the target genes. In this study, we investigated the role and mechanism of miR-10a-5p in CCA.
Human CCA cell lines (CCLP1 and SG-231) were transfected with miR-10a-5p mimic or miR-10a-5p inhibitor. qRT-PCR was performed to detect the miR-10a-5p level. Proliferation, colony formation, and apoptosis were analyzed. Luciferase reporter assay was used to explore the targeting of miR-10a-5p on PTEN. For in vivo tumorigenesis assay, CCLP1 cells with stable knockdown of miR-10a-5p or control CCLP1 cells were injected subcutaneously into the flank of the SCID mice and animals were monitored for tumor growth.
miR-10a-5p expression was significantly upregulated in human CCA cell lines (CCLP1 and SG-231). Inhibition of miR-10a-5p significantly suppressed the proliferation and induced apoptosis in CCLP1 and SG-231. PTEN is a direct target of miR-10a-5p in CCA cells.
Inhibition of miR-10a-5p can decrease CCA cells growth by downregulation of Akt pathway. These results indicate that miR-10a-5p may serve as a potential target for the treatment of CCA and help to develop effective therapeutic strategies.
胆管癌（CCA）是一种预后极差的上皮细胞恶性肿瘤。许多患者在CCA晚期被诊断出来，且未发现危险因素。对于CCA患者的治疗，可用的治疗选择有限。迫切需要开发有效的靶向治疗方法来治疗CCA。微小RNA（miRNA）是负向调节靶基因的小非编码RNA。在本研究中，我们研究了miR-10a-5p在CCA中的作用及机制。
用miR-10a-5p模拟物或miR-10a-5p抑制剂转染人CCA细胞系（CCLP1和SG-231）。进行qRT-PCR检测miR-10a-5p水平。分析细胞增殖、集落形成和凋亡情况。采用荧光素酶报告基因检测法探讨miR-10a-5p对PTEN的靶向作用。对于体内肿瘤发生试验，将稳定敲低miR-10a-5p的CCLP1细胞或对照CCLP1细胞皮下注射到SCID小鼠的侧腹，并监测动物的肿瘤生长情况。
miR-10a-5p在人CCA细胞系（CCLP1和SG-231）中表达显著上调。抑制miR-10a-5p可显著抑制CCLP1和SG-231细胞的增殖并诱导其凋亡。PTEN是CCA细胞中miR-10a-5p的直接靶点。
抑制miR-10a-5p可通过下调Akt通路降低CCA细胞的生长。这些结果表明，miR-10a-5p可能作为治疗CCA的潜在靶点，并有助于制定有效的治疗策略。
