Han Li Li, Yin Xiao Ran, Zhang Shu Qun
Cell Physiol Biochem. 2018;51(3):1179-1192. doi: 10.1159/000495495. Epub 2018 Nov 27.
BACKGROUND/AIMS: Previous studies have confirmed that microRNAs are involved in the metastasis and epithelial-mesenchymal transition (EMT) of malignancies. In this study, we examined whether miR-650 promotes the migration, invasion, and EMT of hepatocellular carcinoma (HCC) cells by targeting the large tumor suppressor kinase 2 gene (LATS2).
qRT-PCR was used to detect expression of miR-650 in HCC tissues and paired normal tissues. MTT and Transwell assay were used to observe the effect of miR-650 on proliferation, migration and invasion of HCC cells. Western blot assay and Immunohistochemistry were performed to demonstrate association between miR-650 expression level and epithelial-mesenchymal transition (EMT) related protein. Mechanistically, Reporter luciferase assay was performed to reveal whether large tumor suppressor kinase 2 (LATS2) was a direct target of miR-650 in HCC cells.
We observed that miR-650 levels were largely up-regulated in HCC tissues, and that the increased expression was closely associated with the adverse clinical features of HCC patients. Additionally, the expression of LATS2, which was identified as a direct target of miR-650, can counteract the effects of miR-650 in HCC. Furthermore, we demonstrated that high miR-650 expression levels and low LATS2 expression levels in tumors may indicate a poor prognosis for HCC patients.
In conclusion, the miR-650/LATS2 pathway may serve as a novel prognostic biomarker and an attractive therapeutic target for HCC patients.
背景/目的:既往研究证实,微小RNA参与恶性肿瘤的转移和上皮-间质转化(EMT)。在本研究中,我们检测了miR-650是否通过靶向大肿瘤抑制激酶2基因(LATS2)促进肝细胞癌(HCC)细胞的迁移、侵袭及EMT。
采用qRT-PCR检测HCC组织及配对正常组织中miR-650的表达。采用MTT和Transwell实验观察miR-650对HCC细胞增殖、迁移和侵袭的影响。进行蛋白质免疫印迹分析和免疫组织化学以证明miR-650表达水平与上皮-间质转化(EMT)相关蛋白之间的关联。从机制上,进行报告荧光素酶实验以揭示大肿瘤抑制激酶2(LATS2)是否为HCC细胞中miR-650的直接靶点。
我们观察到HCC组织中miR-650水平大幅上调,且表达增加与HCC患者的不良临床特征密切相关。此外,被鉴定为miR-650直接靶点的LATS2的表达可抵消miR-650在HCC中的作用。此外,我们证明肿瘤中高miR-650表达水平和低LATS2表达水平可能提示HCC患者预后不良。
总之,miR-650/LATS2通路可能作为HCC患者一种新的预后生物标志物和有吸引力的治疗靶点。