沉睡美人插入突变筛选鉴定了 CNPY2 和 ACTN2 在肝癌肿瘤进展中的促转移作用。

Sleeping Beauty insertional mutagenesis screen identifies the pro-metastatic roles of CNPY2 and ACTN2 in hepatocellular carcinoma tumor progression.

机构信息

State Key Laboratory of Chinese Medicine and Molecular Pharmacology (Incubation), The Hong Kong Polytechnic University Shenzhen Research Institute, Shenzhen, China; Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China; State Key Laboratory of Chemical Biology and Drug Discovery, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China.

出版信息

Biochem Biophys Res Commun. 2021 Feb 19;541:70-77. doi: 10.1016/j.bbrc.2021.01.017. Epub 2021 Jan 19.

DOI:10.1016/j.bbrc.2021.01.017

PMID:33482578

Abstract

A forward genetic Sleeping Beauty (SB) insertional mutagenesis screen, followed by high-throughput transcriptome sequencing, was used to identify driver genes responsible for hepatocellular carcinoma (HCC)-associated metastasis. Using RNA-sequencing (RNA-seq) to identify transposon-endogenous transcriptome fusion genes, the phylogenetic lineage between the parental liver tumor and secondary metastasis can be determined to provide mechanistic insight to genetic changes involved in the metastatic evolution process. In the current study, two novel candidate genes were identified to be potentially involved in HCC-associated metastatic progression, canopy FGF signaling regulator 2 (Cnpy2) and actinin alpha 2 (Actn2). Transposon-Cnpy2 fusion transcripts were identified in both primary liver tumors and lung metastases. Its significant association with clinicopathological characteristics and correlated gene enrichment in metastasis-related mechanisms suggest its potential role in modulating local invasion and angiogenesis. Other known driver genes for human HCC that can also promote metastatic progression include epidermal growth factor receptor (Egfr) and RNA imprinted and accumulated in nucleus (Rian). Metabolic pathway related gene carbamoyl phosphate synthetase (Cps1) was identified to play an important role in early HCC development, while cell junction-related pathway gene Rac family small GTPase 1 (Rac1) was identified to take part in both HCC and pro-metastatic progression. Importantly, actinin alpha 2 (Actn2) was identified exclusively in the secondary metastasis site and its role in HCC-related metastatic process was elucidated using in vitro approaches. ACTN2-overexpression in human liver cancer cells displayed enhanced cellular motility and invasion abilities, indicating its possible function in later stage of metastasis, such as extravasation and lung colonization.

摘要

采用正向遗传的 Sleeping Beauty（SB）插入突变筛选，结合高通量转录组测序，鉴定了与肝细胞癌（HCC）相关转移有关的驱动基因。利用 RNA 测序（RNA-seq）鉴定转座子-内源性转录组融合基因，可以确定亲本肝肿瘤和继发性转移之间的系统发育谱系，为遗传变化在转移进化过程中提供机制见解。在本研究中，鉴定出两个可能参与 HCC 相关转移进展的新候选基因，即 canopy FGF 信号调节因子 2（Cnpy2）和肌动蛋白α 2（Actn2）。转座子-Cnpy2 融合转录本在原发性肝肿瘤和肺转移中均被鉴定出来。其与临床病理特征的显著相关性及其在转移相关机制中的相关基因富集表明，其在调节局部侵袭和血管生成方面具有潜在作用。其他已知的人类 HCC 驱动基因也能促进转移进展，包括表皮生长因子受体（Egfr）和 RNA 印迹和积累核内（Rian）。代谢途径相关基因氨甲酰磷酸合成酶 1（Cps1）被鉴定为在早期 HCC 发展中发挥重要作用，而细胞连接相关途径基因 Rac 家族小 GTP 酶 1（Rac1）被鉴定为参与 HCC 和促转移进展。重要的是，肌动蛋白α 2（Actn2）仅在继发性转移部位被鉴定出来，并通过体外方法阐明了其在 HCC 相关转移过程中的作用。在人肝癌细胞中过表达 ACTN2 可增强细胞迁移和侵袭能力，表明其在转移的后期阶段可能具有功能，如血管外渗和肺定植。