Zhang Bashan, Li Fei, Zhu Zinian, Ding Aijiao, Luo Jintong
Department of Clinical Laboratory, Affiliated Dongguan People's Hospital, Southern Medical University of Dongguan, Dongguan City, Guangdong Province, People's Republic of China.
Cancer Manag Res. 2020 Sep 24;12:8951-8964. doi: 10.2147/CMAR.S252679. eCollection 2020.
Hepatocellular carcinoma (HCC) is a common lethal malignant tumor worldwide. Circular RNAs (circRNAs) have been reported to affect the development of human cancers, including HCC. In this project, we aim to clarify the functional effect of circular CDR1as (circ_CDR1as) on HCC progression.
Quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot is implemented to detect the expression of circ_CDR1as, microRNA (miR)-1287 and Raf-1 proto-oncogene, serine/threonine kinase (Raf1). Cell proliferation is assessed via colony formation and 3-(4, 5)-dimethylthiazole-2-y1)-2, 5-biphenyl tetrazolium bromide (MTT) assays. Cell migration and invasion are measured by Transwell assay. The target relationship between miR-1287 and circ_CDR1as or Raf1 is validated through dual-luciferase reporter assay. The levels of epithelia-mesenchymal transition (EMT) markers and the MEK/ERK signal pathway-related proteins are examined by Western blot. Model in nude mice is constructed to determine the role of circ_CDR1as in vivo.
Expression of circ_CDR1as and Raf1 is elevated, while miR-1287 expression is decreased in HCC. Depletion of circ_CDR1as or Raf1 could inhibit proliferation and metastasis of HCC cells. Besides, circ_CDR1as regulates Raf1 expression by targeting miR-1287. MiR-1287 upregulation or Raf1 depletion could partially counterbalance circ_CDR1as depletion-mediated inhibitory effects on HCC cell behaviors. Moreover, circ_CDR1as depletion represses HCC progression through inactivating MEK/ERK pathway. In addition, circ_CDR1as depletion suppresses tumor growth in vivo via regulating miR-1287/Raf1 pathway.
Circ_CDR1as depletion inhibits HCC cell proliferation and metastasis by miR-1287/Raf1 and MEK/ERK pathways, highlighting a promising molecular target for HCC treatment.
肝细胞癌（HCC）是全球常见的致死性恶性肿瘤。据报道，环状RNA（circRNAs）会影响包括HCC在内的人类癌症的发展。在本项目中，我们旨在阐明环状CDR1as（circ_CDR1as）对HCC进展的功能影响。
采用定量实时聚合酶链反应（qRT-PCR）或蛋白质免疫印迹法检测circ_CDR1as、微小RNA（miR）-1287和原癌基因Raf-1丝氨酸/苏氨酸激酶（Raf1）的表达。通过集落形成和3-(4,5)-二甲基噻唑-2-基)-2,5-联苯四氮唑溴盐（MTT）试验评估细胞增殖。采用Transwell试验检测细胞迁移和侵袭能力。通过双荧光素酶报告基因试验验证miR-1287与circ_CDR1as或Raf1之间的靶向关系。通过蛋白质免疫印迹法检测上皮-间质转化（EMT）标志物和MEK/ERK信号通路相关蛋白的水平。构建裸鼠模型以确定circ_CDR1as在体内的作用。
HCC中circ_CDR1as和Raf1的表达升高，而miR-1287的表达降低。circ_CDR1as或Raf1的缺失可抑制HCC细胞的增殖和转移。此外，circ_CDR1as通过靶向miR-1287调节Raf1的表达。miR-1287上调或Raf1缺失可部分抵消circ_CDR1as缺失介导的对HCC细胞行为的抑制作用。此外，circ_CDR1as缺失通过使MEK/ERK通路失活来抑制HCC进展。此外，circ_CDR1as缺失通过调节miR-1287/Raf1通路抑制体内肿瘤生长。
circ_CDR1as缺失通过miR-1287/Raf1和MEK/ERK通路抑制HCC细胞增殖和转移，突出了其作为HCC治疗有前景的分子靶点的作用。
