Wang Ping, Lv Long
Department of Endocrinology, People's Hospital of Gaochun, Gaochun, Nanjing, 211300, Jiangsu Province, China.
Department of General Surgery, People's Hospital of Gaochun, Gaochun, Nanjing, 211300, Jiangsu Province, China.
Oncotarget. 2016 Dec 6;7(49):81367-81376. doi: 10.18632/oncotarget.13229.
KRT19 was identified as one of the key biomarkers for distinguishing cholangiocarcinoma (CCA) and hepatocellular carcinoma. The detailed role of miRNAs involved in the oncogenic incident of KRT19 was poor investigated.
Based on prediction and validation, miR-26a was inversely correlated with KRT19 in patients' tissues samples and biopsies. Ectopic expression of miR-26a dramatically suppressed cell proliferation and tumor growth in vitro and in vivo. Knock-down miR-26a could induce an increasing population of SP cells by promoting KRT19 expression. The KRT19 was also suppressed via directly binding at 3'UTR region by miR-26a.
Bioinformatics prediction was first applied to screening the potential miRNA involved. RT-PCR, Immunohistochemistry and Western blot were used to examine the expression of miRNAs and candidate genes in 65 pairs of cholangiocarcinoma. The loss-and gain-function assay was employed to detect the role of certain miRNA in vitro and vivo. Side-population (SP) cells were detected and sorted by flow cytometry.
Aberrant decreased miR-26a could promote cell proliferation by regulating KRT19 which play important roles in the pathogenesis of CCA.
KRT19被鉴定为区分胆管癌(CCA)和肝细胞癌的关键生物标志物之一。关于参与KRT19致癌事件的微小RNA(miRNA)的详细作用研究较少。
基于预测和验证,miR-26a在患者组织样本和活检中与KRT19呈负相关。miR-26a的异位表达在体外和体内均显著抑制细胞增殖和肿瘤生长。敲低miR-26a可通过促进KRT19表达诱导侧群(SP)细胞数量增加。miR-26a还通过直接结合KRT19的3'非翻译区(UTR)来抑制其表达。
首先应用生物信息学预测筛选潜在的相关miRNA。采用逆转录-聚合酶链反应(RT-PCR)、免疫组织化学和蛋白质免疫印迹法检测65对胆管癌组织中miRNA和候选基因的表达。采用功能缺失和功能获得实验检测特定miRNA在体外和体内的作用。通过流式细胞术检测和分选侧群(SP)细胞。
miR-26a异常降低可通过调节KRT19促进细胞增殖,而KRT19在CCA发病机制中起重要作用。