CPSF6 将可变多聚腺苷酸化与肝癌进展中的代谢适应联系起来。
CPSF6 links alternative polyadenylation to metabolism adaption in hepatocellular carcinoma progression.
机构信息
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, 200240, China.
Department of Integrated Traditional Chinese and Western Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, 200030, China.
出版信息
J Exp Clin Cancer Res. 2021 Mar 1;40(1):85. doi: 10.1186/s13046-021-01884-z.
BACKGROUND
Alternative polyadenylation (APA) is an important mechanism of gene expression regulation through generation of RNA isoforms with distinct 3' termini. Increasing evidence has revealed that APA is actively involved in development and disease, including hepatocellular carcinoma (HCC). However, how APA functions in tumor formation and progression remains elusive. In this study, we investigated the role of cleavage factor I (CFIm) subunit CPSF6 in human hepatocellular carcinoma (HCC).
METHODS
Expression levels of CPSF6 in clinical tissues and cell lines were determined by qRT-PCR and western blot. Functional assays, including the cell number, MTT, colony formation and transwell, were used to determine the oncogenic role of CPSF6 in HCC. Animal experiments were used to determine the role of CPSF6 in HCC tumorigenicity in vivo. Deep sequencing-based 3 T-seq was used to profile the transcriptome-wide APA sites in both HCC cells and CPSF6 knockdown HCC cells. The function of CPSF6-affected target NQO1 with distinct 3'UTRs was characterized by metabolism assays.
RESULTS
We observed CPSF6 was upregulated in HCC and the high expression of CPSF6 was associated with poor prognosis in patients. Overexpression of CPSF6 promoted proliferation, migration and invasion of HCC cells in vitro and in vivo. Transcriptome-wide APA profiling analysis indicated that high expression of CPSF6 promoted the favorable usage of the proximal poly(A) site in the 3'UTR of NQO1. We demonstrated CPSF6-induced tumorigenic activities were mediated by the NQO1 isoform with short 3'UTR. Furthermore, we found that CPSF6 induced metabolic alterations in liver cells through NQO1.
CONCLUSION
CPSF6 plays a critical role in HCC progression by upregulating NQO1 expression through APA. These findings provide evidence to demonstrate that APA of NQO1 contributes to HCC progression and may have implications for developing new therapeutic strategy against this disease.
背景
可变多聚腺苷酸化（APA）是一种通过生成具有不同 3'末端的 RNA 异构体来调节基因表达的重要机制。越来越多的证据表明，APA 积极参与了发育和疾病，包括肝细胞癌（HCC）。然而，APA 如何在肿瘤形成和进展中发挥作用仍不清楚。在这项研究中，我们研究了剪接因子 I（CFIm）亚基 CPSF6 在人肝细胞癌（HCC）中的作用。
方法
通过 qRT-PCR 和 Western blot 测定临床组织和细胞系中 CPSF6 的表达水平。细胞计数、MTT、集落形成和 Transwell 等功能测定用于确定 CPSF6 在 HCC 中的致癌作用。动物实验用于确定 CPSF6 在 HCC 肿瘤发生中的体内作用。基于深度测序的 3T-seq 用于分析 HCC 细胞和 CPSF6 敲低 HCC 细胞中转录组范围的 APA 位点。通过代谢测定来表征具有不同 3'UTR 的 CPSF6 影响的靶标 NQO1 的功能。
结果
我们观察到 CPSF6 在 HCC 中上调，并且 CPSF6 的高表达与患者的预后不良相关。CPSF6 的过表达促进了 HCC 细胞在体外和体内的增殖、迁移和侵袭。转录组范围的 APA 谱分析表明，CPSF6 的高表达促进了 NQO1 3'UTR 中近端 poly(A) 位点的有利使用。我们证明 CPSF6 诱导的肿瘤发生活性是通过 NQO1 的短 3'UTR 同工型介导的。此外，我们发现 CPSF6 通过 NQO1 诱导肝细胞中的代谢改变。
结论
CPSF6 通过 APA 上调 NQO1 的表达在 HCC 进展中发挥关键作用。这些发现为 NQO1 的 APA 促进 HCC 进展提供了证据，并可能为开发针对这种疾病的新治疗策略提供依据。
Zotero 插件