Knockdown of ubiquitin-specific peptidase 39 inhibited the growth of osteosarcoma cells and induced apoptosis in vitro.
Knockdown of ubiquitin-specific peptidase 39 inhibited the growth of osteosarcoma cells and induced apoptosis in vitro.
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DOI：
10.1186/s40659-017-0121-z
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发表时间：
2017-04-12
影响因子：
6.7
通讯作者：
Min D
中科院分区：
文献类型：
--
作者：
Gan Z;Han K;Lin S;Hu H;Shen Z;Min D
泛素特异性肽酶39（USP 39）是成熟剪接体复合物组装中的一个重要因子，在几种癌症中有异常表达。然而，其对人骨肉瘤的作用及其机制尚未完全阐明。根据公开可用的Oncomine数据库的数据集，与人类正常组织相比，骨肉瘤癌组织中USP 39的mRNA和DNA拷贝增加。进一步的蛋白质印迹分析也证明了USP 39在三种不同的骨肉瘤细胞中的异常内源性表达。然后设计慢病毒介导的短发夹状RNA（short hairpin RNA，shRNA）沉默人骨肉瘤细胞系U2 OS中USP 39，检测USP 39沉默对细胞增殖、集落形成、细胞周期分布和凋亡的影响。U2 OS细胞中USP 39表达的敲低显著降低细胞增殖，损害集落形成能力。进一步分析表明，USP 39的抑制通过p21依赖的方式将细胞周期进程阻滞在G2/M期。Annexin V/7-AAD染色结果表明，USP 39基因的敲低可通过PARP裂解促进U2 OS细胞凋亡。这些结果揭示了USP 39在调节癌细胞有丝分裂中的关键作用，并表明USP 39对骨肉瘤肿瘤发生至关重要。
Ubiquitin specific peptidase 39 (USP39), an essential factor in the assembly of the mature spliceosome complex, has an aberrant expression in several cancer. However, its function and the corresponding mechanism on human osteosarcoma has not been fully explored yet. The mRNA and DNA copies of USP39 were increased in osteosarcoma cancer tissues compared with the one in human normal tissues according to datasets from the publicly available Oncomine database. A further western blot analysis also demonstrated an aberrant endogenous expression of USP39 in three different osteosarcoma cells. Then lentivirus-mediated short hairpin RNA (shRNA) was designed to silence USP39 in human osteosarcoma cell line U2OS, which is used to test the impact of USP39-silencing on cellular proliferation, colony formation, cell cycle distribution and apoptosis. Knockdown of USP39 expression in U2OS cell significantly decreased cell proliferation, impaired colony formation ability. A further analysis indicated suppression of USP39 arrested cell cycle progression at G2/M phase via p21 dependent way. In addition, the results of Annexin V/7-AAD staining suggested the knockdown of USP39 could promote U2OS cell apoptosis through PARP cleavage. These results uncover the critical role of USP39 in regulating cancer cell mitosis and indicate USP39 is critical for osteosarcoma tumorigenesis.