Dong Shuilin, Wang Wei, Liao Zhibin, Fan Yawei, Wang Qi, Zhang Lei
Hepatic Surgery Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Hubei Key Laboratory of Hepato-Pancreato-Biliary Diseases, Wuhan, Hubei, China.
J Gene Med. 2023 Apr;25(4):e3477. doi: 10.1002/jgm.3477. Epub 2023 Feb 14.
There have been many reports of long non-coding RNAs (lncRNAs) in tumors, and abnormally expressed lncRNA is closely related to hepatocellular carcinoma (HCC). The mechanism of LINC00607 in HCC has not been reported.
We utilized qPCR to evaluate the RNA expression level. The mechanism of MYC binding to the LINC00607 promoter was revealed through chromatin immunoprecipitation assay and dual luciferase reporter assay. The proliferation and invasive ability were evaluated by CCK-8 and transwell assays. The relation between LINC00607 and miR-584-3p was assessed by RNA immunoprecipitation assay and dual luciferase reporter assay. The level of ROCK1 was evaluated by qPCR and western blot.
In this research, we found that the expression of LINC00607 was higher in HCC tissues when compared with that in the adjacent non-tumor tissues. Meanwhile, MYC was observed to interact with the LINC00607 promoter, leading to the upregulation of LINC00607 in HCC. We further revealed that LINC00607 functioned as a sponge for miR-584-3p. Cell proliferation and migration assays showed that miR-584-3p may inhibit the HCC progression. Moreover, we found that the miR-584-3p inhibitor could reverse the effects of LINC00607 downregulation in HCC through rescue experiments. Through verification, miR-584-3p bound to the 3' UTR of ROCK1 to downregulate its expression.
LINC00607 regulated by MYC can promote the proliferation, migration and invasion of HCC cells through the miR-584-3p/ROCK1 axis.
肿瘤中长链非编码RNA（lncRNAs）已有诸多报道，且异常表达的lncRNA与肝细胞癌（HCC）密切相关。LINC00607在HCC中的作用机制尚未见报道。
我们运用qPCR评估RNA表达水平。通过染色质免疫沉淀实验和双荧光素酶报告基因实验揭示MYC与LINC00607启动子的结合机制。采用CCK-8和Transwell实验评估细胞增殖和侵袭能力。通过RNA免疫沉淀实验和双荧光素酶报告基因实验评估LINC00607与miR-584-3p之间的关系。运用qPCR和蛋白质免疫印迹法评估ROCK1的水平。
在本研究中，我们发现与癌旁非肿瘤组织相比，LINC00607在HCC组织中的表达更高。同时，观察到MYC与LINC00607启动子相互作用，导致LINC00607在HCC中上调。我们进一步揭示LINC00607作为miR-584-3p的海绵发挥作用。细胞增殖和迁移实验表明，miR-584-3p可能抑制HCC进展。此外，通过拯救实验我们发现miR-584-3p抑制剂可逆转LINC00607下调对HCC的影响。经证实，miR-584-3p与ROCK1的3'UTR结合以下调其表达。
由MYC调控的LINC00607可通过miR-584-3p/ROCK1轴促进HCC细胞的增殖、迁移和侵袭。
