长链非编码 RNA-Gm9866 通过靶向 Fam98b 激活 TGFβ/Smad 信号通路促进肝纤维化。
LncRNA-Gm9866 promotes liver fibrosis by activating TGFβ/Smad signaling via targeting Fam98b.
机构信息
Department of Gastroenterology, The First Affiliated Hospital of Guangxi Medical University, No. 6, Shuangyong Road, Nanning, 530021, Guangxi, China.
Department of Gastroenterology, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan, China.
出版信息
J Transl Med. 2023 Nov 2;21(1):778. doi: 10.1186/s12967-023-04642-1.
OBJECTIVE
The exact mechanism and target molecules of liver fibrosis have remained largely elusive. Here, we investigated the role of long noncoding RNA Gm9866(lncRNA-Gm9866) on liver fibrosis.
METHODS
The transcription of lncRNA-Gm9866 in activated cells and mouse fibrotic livers was determined by quantitative polymerase chain reaction (qRT-PCR). The effects of lentivirus-mediated knockdown or overexpression of lncRNA-Gm9866 in liver fibrosis were examined in vitro and in vivo. Furthermore, bioinformatics analysis, cell samples validation, fluorescence in situ hybridization (FISH) co-localization, RNA binding protein immunoprecipitation (RIP), actinomycin D test and Western blot (WB) were carried out to explore the potential mechanism of lncRNA-Gm9866.
RESULTS
The expression of α-smooth muscle actin (α-SMA), Collagen I (COL-1) and lncRNA-Gm9866 were significantly increased in tissues and cells. Overexpressing lncRNA-Gm9866 promoted the activation of hepatic stellate cells (HSCs). Silencing lncRNA-Gm9866 inhibited the activation of HSCs and transforming growth factor-β1 (TGFβ1) induced fibrosis. Overexpressing lncRNA-Gm9866 promoted hepatocytes (HCs) apoptosis and the expression of pro-fibrogenic genes, inhibited the proliferation and migration of HCs. Knockdown of lncRNA-Gm9866 inhibited the apoptosis of HCs, the expression of pro-fibrogenic genes, TGFβ1 induced fibrosis and the occurrence of carbon tetrachloride (CCl4)-induced liver fibrosis, and promoted the proliferation and migration of HCs. Mechanistically, lncRNA-Gm9866 may directly bine with Fam98b. Silencing Fam98b in stably overexpressing lncRNA-Gm9866 cell lines reversed the increase of pro-fibrogenic genes and pro-apoptotic genes, fibrosis related pathway protein TGFβ1, Smad2/3, p-Smad2/3 and Notch3 induced by overexpressing lncRNA-Gm9866.
CONCLUSIONS
LncRNA-Gm9866 may regulate TGFβ/Smad and Notch pathways by targeting Fam98b to regulate liver fibrosis. LncRNA-Gm9866 may be a new target for diagnosis and treatment of liver fibrosis.
目的
肝纤维化的确切机制和靶分子在很大程度上仍未被发现。本研究旨在探讨长链非编码 RNA Gm9866（lncRNA-Gm9866）在肝纤维化中的作用。
方法
采用实时定量聚合酶链反应（qRT-PCR）检测激活细胞和小鼠纤维化肝脏中 lncRNA-Gm9866 的转录。在体外和体内研究了慢病毒介导的 lncRNA-Gm9866 敲低或过表达对肝纤维化的影响。此外，进行了生物信息学分析、细胞样本验证、荧光原位杂交（FISH）共定位、RNA 结合蛋白免疫沉淀（RIP）、放线菌素 D 试验和 Western blot（WB），以探讨 lncRNA-Gm9866 的潜在机制。
结果
α-平滑肌肌动蛋白（α-SMA）、胶原 I（COL-1）和 lncRNA-Gm9866 的表达在组织和细胞中均显著增加。过表达 lncRNA-Gm9866 促进肝星状细胞（HSCs）的激活。沉默 lncRNA-Gm9866 抑制 HSCs 激活和转化生长因子-β1（TGFβ1）诱导的纤维化。过表达 lncRNA-Gm9866 促进肝细胞（HCs）凋亡和促纤维化基因的表达，抑制 HCs 的增殖和迁移。敲低 lncRNA-Gm9866 抑制 HCs 的凋亡、促纤维化基因的表达、TGFβ1 诱导的纤维化和四氯化碳（CCl4）诱导的肝纤维化，并促进 HCs 的增殖和迁移。机制上，lncRNA-Gm9866 可能直接与 Fam98b 结合。在稳定过表达 lncRNA-Gm9866 的细胞系中沉默 Fam98b，逆转了过表达 lncRNA-Gm9866 引起的促纤维化基因和促凋亡基因、纤维化相关通路蛋白 TGFβ1、Smad2/3、p-Smad2/3 和 Notch3 的增加。
结论
lncRNA-Gm9866 可能通过靶向 Fam98b 调节 TGFβ/Smad 和 Notch 通路来调节肝纤维化。lncRNA-Gm9866 可能成为肝纤维化诊断和治疗的新靶点。
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