Gao Jun, Li Enliang, Liu Weiwei, Yang Qingping, Xie Chunyan, Ai Jiyuan, Zhou Fan, Liao Wenjun, Wu Linquan
Department of Hepatobiliary and Pancreatic Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China.
Department of Assisted Reproductive, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China.
Onco Targets Ther. 2020 Aug 25;13:8615-8627. doi: 10.2147/OTT.S258715. eCollection 2020.
This study aimed to investigate the functions of the circular RNA circMYLK (hsa_circ_0002768) in the development of hepatocellular carcinoma (HCC) and to identify the underlying mechanisms of the circMYLK/miR29a/KMT5C axis.
Quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to explore the expressions of circMYLK, miR-29a and KMT5C in HCC tissues and cells. A potential miRNA (miR-29a) regulated by circMYLK was also explored, and the target relationship between miR-29a and KMT5C was confirmed. FISH, qRT-PCR, Western blotting, and dual-luciferase reporter assays were used to examine the circMYLK/miR29a/KMT5C signaling pathways involved in HCC development. Additionally, HCC cells were implanted into nude mice subcutaneously to test the role of circMYLK in tumor growth.
circMYLK was determined to be significantly upregulated in HCC tissues and cells. Suppression of circMYLK repressed HCC cell proliferation, migration, and invasion while increasing apoptosis. In addition, FISH, qRT-PCR, and Western blotting, as well as dual-luciferase reporter assays, revealed that circMYLK could bind to miR-29a. In rescue experiments, miR-29a had the potential to eliminate the inhibitory effect of circMYLK knockdown in HCC. Moreover, miR-29a was found to target the gene, which was positively regulated by circMYLK. Finally, a nude mouse tumorigenicity assay showed that injection of circMYLK siRNA into nude mice drastically suppressed xenograft tumor formation in vivo.
Our current study demonstrated that circMYLK promotes HCC progression by acting as a competing endogenous RNA of miR-29a, which regulates the downstream oncogene .
本研究旨在探讨环状RNA circMYLK（hsa_circ_0002768）在肝细胞癌（HCC）发生发展中的作用，并确定circMYLK/miR29a/KMT5C轴的潜在机制。
采用定量实时聚合酶链反应（qRT-PCR）检测circMYLK、miR-29a和KMT5C在HCC组织和细胞中的表达。还探究了受circMYLK调控的潜在微小RNA（miR-29a），并证实了miR-29a与KMT5C之间的靶向关系。采用荧光原位杂交（FISH）、qRT-PCR、蛋白质免疫印迹法和双荧光素酶报告基因检测法，研究参与HCC发生发展的circMYLK/miR29a/KMT5C信号通路。此外，将HCC细胞皮下接种到裸鼠体内，以检测circMYLK在肿瘤生长中的作用。
circMYLK在HCC组织和细胞中显著上调。抑制circMYLK可抑制HCC细胞增殖、迁移和侵袭，同时增加细胞凋亡。此外，FISH、qRT-PCR、蛋白质免疫印迹法以及双荧光素酶报告基因检测法显示，circMYLK可与miR-29a结合。在拯救实验中，miR-29a有可能消除circMYLK敲低对HCC的抑制作用。此外，发现miR-29a靶向该基因，该基因受circMYLK正向调控。最后，裸鼠致瘤性实验表明，向裸鼠注射circMYLK siRNA可显著抑制体内异种移植瘤的形成。
我们目前的研究表明，circMYLK作为miR-29a的竞争性内源性RNA促进HCC进展，miR-29a调节下游癌基因。
