Tang Jianmin, Wu Zheng, Wang Xiaohang, Hou Yanli, Bai Yongrui, Tian Ye
Department of Radiotherapy and Oncology, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
Institute of Radiotherapy and Oncology, Soochow University, Suzhou 215004, China.
Cells. 2022 Oct 28;11(21):3407. doi: 10.3390/cells11213407.
The role of hypoxia-regulated long non-coding RNA (lncRNA) in the development of head and neck squamous cell carcinoma (HNSCC) remains to be elucidated. In the current study, we initially screened hypoxia-regulated lncRNA in HNSCC cells by RNA-seq, before focusing on the rarely annotated lncRNA USP2 antisense RNA 1 (USP2-AS1). We determined that USP2-AS1 is a direct target of HIF1α and is remarkably elevated in HNSCC compared with matched normal tissues. Patients with a higher level of USP2-AS1 suffered a poor prognosis. Next, loss- and gain-of-function assays revealed that USP2-AS1 promoted cell proliferation and invasion in vitro and in vivo. Mechanically, RNA pulldown and LC-MS/MS demonstrated that the E3 ligase DDB1- and CUL4-associated factor 13 (DCAF13) is one of the binding partners to USP2-AS1 in HNSCC cells. In addition, we assumed that USP2-AS1 regulates the activity of DCAF13 by targeting its substrate ATR. Moreover, the knockdown of DCAF13 restored the elevated cell proliferation and growth levels achieved by USP2-AS1 overexpression. Altogether, we found that lncRNA USP2-AS1 functions as a HIF1α-regulated oncogenic lncRNA and promotes HNSCC cell proliferation and growth by interacting and modulating the activity of DCAF13.
缺氧调节的长链非编码RNA（lncRNA）在头颈部鳞状细胞癌（HNSCC）发生发展中的作用仍有待阐明。在本研究中，我们首先通过RNA测序在HNSCC细胞中筛选缺氧调节的lncRNA，之后聚焦于注释很少的lncRNA泛素特异性蛋白酶2反义RNA 1（USP2-AS1）。我们确定USP2-AS1是缺氧诱导因子1α（HIF1α）的直接靶点，与配对的正常组织相比，其在HNSCC中显著上调。USP2-AS1水平较高的患者预后较差。接下来，功能缺失和功能获得实验表明，USP2-AS1在体外和体内均促进细胞增殖和侵袭。机制上，RNA下拉和液相色谱-串联质谱分析表明，E3泛素连接酶损伤特异性DNA结合蛋白1和Cullin4相关因子13（DCAF13）是HNSCC细胞中USP2-AS1的结合伙伴之一。此外，我们推测USP2-AS1通过靶向DCAF13的底物共济失调毛细血管扩张症和Rad3相关蛋白（ATR）来调节DCAF13的活性。而且，敲低DCAF13可恢复USP2-AS1过表达所导致的细胞增殖和生长水平升高。总之，我们发现lncRNA USP2-AS1作为一种受HIF1α调节的致癌lncRNA，通过与DCAF13相互作用并调节其活性来促进HNSCC细胞的增殖和生长。
