Hepatobiliary and Pancreatic Surgery Department, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, China.
J Cell Mol Med. 2022 May;26(9):2505-2519. doi: 10.1111/jcmm.16845. Epub 2022 Mar 29.
MiR-589-5p could promote liver cancer, but the specific mechanisms are largely unknown. This study examined the role and mechanisms of miR-589-5p in liver cancer. The expressions of miR-589-5p, METTL3 and m6A in liver cancers were determined by RT-qPCR. The relationship between miR-589-5p and METTL3-mediated m6A methylation was examined by m6A RNA immunoprecipitation. After transfection, the viability, migration, invasion and expressions of METTL3 and miR-589-5p in liver cancer cells were detected by CCK-8, wound-healing, transwell and RT-qPCR. After the xenograft tumour was established in mice, the tumour volume was determined and the expressions of METTL3, miR-589-5p, MMP-2, TIMP-2, E-cadherin, N-cadherin and Vimentin in tumour tissue were detected by RT-qPCR and Western blotting. In vitro study showed that miR-589-5p and METTL3 were highly expressed in liver cancer. METTL3 was positively correlated with miR-589-5p. METTL3 up-regulated the expression of miR-589-5p and promoted the maturation of miR-589-5p. Overexpressed miR-589-5p and METTL3 promoted the viability, migration and invasion of liver cancer cells, while the effects of silencing miR-589-5p and METTL3 on the cells were the opposite. The effects of METTL3 overexpression and silencing were reversed by miR-589-5p inhibitor and mimic, respectively. In vivo study showed that METLL3 silencing inhibited the growth of xenograft tumour and the expressions of METTL3, MMP-2, N-cadherin and Vimentin, promoted the expressions of TIMP-2 and E-cadherin, while miR-589-5p mimic caused the opposite results and further reversed the effects of METLL3 silencing. In summary, this study found that METTL3-mediated maturation of miR-589-5p promoted the malignant development of liver cancer.
miR-589-5p 可能促进肝癌，但具体机制尚不清楚。本研究探讨了 miR-589-5p 在肝癌中的作用和机制。通过 RT-qPCR 测定肝癌中 miR-589-5p、METTL3 和 m6A 的表达。通过 m6A RNA 免疫沉淀检测 miR-589-5p 与 METTL3 介导的 m6A 甲基化之间的关系。转染后，通过 CCK-8、划痕愈合、transwell 和 RT-qPCR 检测肝癌细胞的活力、迁移和侵袭以及 METTL3 和 miR-589-5p 的表达。在小鼠中建立异种移植瘤后，测定肿瘤体积，并通过 RT-qPCR 和 Western blot 检测肿瘤组织中 METTL3、miR-589-5p、MMP-2、TIMP-2、E-cadherin、N-cadherin 和 Vimentin 的表达。体外研究表明，miR-589-5p 和 METTL3 在肝癌中高表达。METTL3 与 miR-589-5p 呈正相关。METTL3 上调 miR-589-5p 的表达并促进 miR-589-5p 的成熟。过表达 miR-589-5p 和 METTL3 促进肝癌细胞的活力、迁移和侵袭，而沉默 miR-589-5p 和 METTL3 对细胞的作用则相反。METTL3 过表达和沉默的作用分别被 miR-589-5p 抑制剂和模拟物逆转。体内研究表明，METLL3 沉默抑制异种移植瘤的生长和 METTL3、MMP-2、N-cadherin 和 Vimentin 的表达，促进 TIMP-2 和 E-cadherin 的表达，而 miR-589-5p 模拟物则产生相反的结果，并进一步逆转了 METLL3 沉默的作用。综上所述，本研究发现 METTL3 介导的 miR-589-5p 成熟促进了肝癌的恶性发展。
