Teng Xiangnan, Shang Jin, Du Lingyao, Huang Wei, Wang Yonghong, Liu Miao, Ma Yuanji, Wang Ming, Tang Hong, Bai Lang
Center of Infectious Diseases, West China Hospital, Sichuan University, Chengdu, China.
Liver Transplantation Center and HBP Surgery, Sichuan Clinical Research Center for Cancer Sichuan Cancer Hospital & Institute, Sichuan Cancer Center, Affiliated Cancer Hospital of University of Electronic Science and Technology of China, Chengdu, China.
J Gastroenterol Hepatol. 2024 Dec;39(12):2892-2902. doi: 10.1111/jgh.16735. Epub 2024 Sep 29.
Epithelial-mesenchymal transition (EMT) is central to HCC metastasis, in which RNA-binding proteins (RBPs) play a key role.
To explore the role of RBPs in metastasis of hepatocellular carcinoma (HCC), whole transcriptome sequencing was conducted to identify differential RBPs between HCC with metastasis and HCC without metastasis. The influence of RBPs on metastasis of HCC was verified by in vitro and in vivo experiments. The interaction of RBPs with non-coding RNAs was evaluated by RNA immunoprecipitation and pull-down assays. RNA sequencing, whole-genome sequencing, and alternative splicing analysis were further performed to clarify post-transcriptional regulation mechanisms.
Whole transcriptome sequencing results showed that expression of thioredoxin (Trx) was significantly upregulated in HCC patients with metastasis. Trx was also found to be associated with poor prognosis in HCC patients. Overexpression of Trx could promote migration and invasion of HCC cells in vitro and increase the rate of lung metastasis of HCC cells in vivo. Moreover, binding assays showed that Trx could bind to LINC00152. As a result, LINC00152 was verified to determine the pro-metastasis function of Trx by knockdown assay. Furthermore, we revealed that Trx could regulate metastasis-associated alternative splicing program. Specifically, angiopoietin 1 (ANGPT1) was the splicing target; the splicing isoform switching of ANGPT1 could activate the PI3K-Akt pathway, upregulate EMT-associated proteins, and promote migration and invasion of HCC cells.
We found that Trx could interact with LINC00152 and promote HCC metastasis via regulating alternative splicing, indicating that Trx may serve as a novel therapeutic target for HCC treatment.
上皮-间质转化（EMT）是肝癌转移的核心过程，其中RNA结合蛋白（RBPs）发挥关键作用。
为探究RBPs在肝细胞癌（HCC）转移中的作用，进行全转录组测序以鉴定有转移和无转移的HCC之间的差异RBPs。通过体外和体内实验验证RBPs对HCC转移的影响。通过RNA免疫沉淀和下拉实验评估RBPs与非编码RNA的相互作用。进一步进行RNA测序、全基因组测序和可变剪接分析以阐明转录后调控机制。
全转录组测序结果显示，硫氧还蛋白（Trx）的表达在有转移的HCC患者中显著上调。还发现Trx与HCC患者的不良预后相关。Trx的过表达可促进体外HCC细胞的迁移和侵袭，并增加体内HCC细胞的肺转移率。此外，结合实验表明Trx可与LINC00152结合。结果，通过敲低实验验证LINC00152可决定Trx的促转移功能。此外，我们揭示Trx可调节与转移相关的可变剪接程序。具体而言，血管生成素1（ANGPT1）是剪接靶点；ANGPT1的剪接异构体转换可激活PI3K-Akt途径，上调与EMT相关的蛋白，并促进HCC细胞的迁移和侵袭。
我们发现Trx可与LINC00152相互作用，并通过调节可变剪接促进HCC转移，表明Trx可能作为HCC治疗的新靶点。
