Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, 350025, Fujian Province, China.
Fuzhong Clinical Medical College of Fujian Medical University, Fuzhou, 362002, Fujian Province, China.
Sci Rep. 2022 Dec 8;12(1):21199. doi: 10.1038/s41598-022-25738-6.
miR-34a targeting on Smad4 plays important role in TGF-β1 pathway which is a dominant factor for balancing collagen production and degradation in hepatic stellate cells. TGF-β1/Smad4 regulated collagen deposition is a hallmark of hepatic fibrosis. The potential regulation on miR-34a by LncRNAs in hepatic stellate cells (HSCs) is still reserved to be revealed. In current study, it was hypothesized that a miR-34a interactor, lncRNA CCAT2 may regulate TGF-β1 pathway in liver fibrotic remodeling. The interaction between CCAT2 and miR-34a-5p was checked by dual luciferase assay. the effects of CCAT2 and miR-34a-5p on cell proliferation and apoptosis were verified by MTT assay, colony formation assay, and flow cytometry assay. Dual luciferase activity showed CCAT2 are targets of miR-34a-5p. Sh-CCAT2 transfection prohibit HSCs proliferation and induce HSCs apoptosis, also inhibited ECM protein synthesis in HSCs. Decreased miR-34a-5p enhanced HSCs proliferation, blocked HSCs apoptosis and promoted ECM protein production. miR-34a-5p inhibitor undo protective regulation of sh-CCAT2 in liver fibrosis. Furthermore, clinical investigation showed that CCAT2 and Smad4 expression level were significantly induced, while miR-34a-5p was significantly decreased in HBV related liver fibrosis serum. In conclusion, activated HSCs via TGF-β1/Smad4 signaling pathway was successfully alleviated by CCAT2 inhibition through miR-34a-5p elevation.
miR-34a 通过靶向 Smad4 在 TGF-β1 通路中发挥重要作用,TGF-β1/Smad4 调节胶原沉积是肝纤维化的标志。lncRNA 在肝星状细胞(HSCs)中对 miR-34a 的潜在调节作用仍有待揭示。在本研究中,假设 miR-34a 的相互作用物 lncRNA CCAT2 可能调节肝纤维化重塑中的 TGF-β1 通路。通过双荧光素酶报告基因检测验证 CCAT2 与 miR-34a-5p 的相互作用。通过 MTT 检测、集落形成检测和流式细胞术检测验证 CCAT2 和 miR-34a-5p 对细胞增殖和凋亡的影响。双荧光素酶活性显示 CCAT2 是 miR-34a-5p 的靶标。Sh-CCAT2 转染可抑制 HSCs 增殖并诱导 HSCs 凋亡,同时抑制 HSCs 细胞外基质蛋白合成。miR-34a-5p 下调增强 HSCs 增殖,阻止 HSCs 凋亡并促进 ECM 蛋白产生。miR-34a-5p 抑制剂消除了 sh-CCAT2 在肝纤维化中的保护调节作用。此外,临床研究表明,HBV 相关肝纤维化血清中 CCAT2 和 Smad4 表达水平显著升高,而 miR-34a-5p 表达水平显著降低。总之,通过上调 miR-34a-5p 抑制 CCAT2 可减轻 TGF-β1/Smad4 信号通路激活的 HSCs。