Zhao Jie, Wei Xiao-Li, Jia Yong-Sheng, Zheng Jian-Quan
Beijing Institute of Pharmacology and Toxicology, Taiping Road 27, Beijing 100850, China.
Eur J Pharmacol. 2008 Jan 28;579(1-3):50-7. doi: 10.1016/j.ejphar.2007.10.008. Epub 2007 Oct 13.
Overexpression of human ether-à-go-go (eag) related gene (herg) contributes to the progression and metastasis of a variety of tumors of different histogenesis, which implies that the herg gene could provide a promising target on tumor therapy. In the present study, plasmid-mediated expression of shRNA-herg1 and shRNA-herg1/1b was employed to silence the herg gene expression in human neuroblastoma SH-SY5Y cell lines. The inhibition of the target gene expression was confirmed by RT-PCR and Western blot. It was found that shRNA-herg1 or shRNA-herg1/1b depressed the cellular growth rate, inhibited cell viability and reduced colony formation of SH-SY5Y cells. The flow cytometry assay revealed that SH-SY5Y cells were retarded in G0-G1 after herg1 or herg1/1b gene was silenced by shRNA-herg1 or shRNA-herg1/1b. In vivo, intra-tumor injection of shRNA-herg1/1b inhibited the growth of SH-SY5Y tumors inoculated subcutaneously in nude mice. The result suggested that the herg played an important role in regulating the growth and proliferation of SH-SY5Y cells. The block of the HERG channel might be a potential therapeutic strategy for neuroblastoma and some other tumors with overexpression of the herg gene.
人醚-去-去(eag)相关基因(herg)的过表达促进了多种不同组织发生的肿瘤的进展和转移,这表明herg基因可能为肿瘤治疗提供一个有前景的靶点。在本研究中,采用质粒介导的shRNA-herg1和shRNA-herg1/1b表达来沉默人神经母细胞瘤SH-SY5Y细胞系中的herg基因表达。通过RT-PCR和蛋白质印迹法证实了靶基因表达的抑制。发现shRNA-herg1或shRNA-herg1/1b降低了SH-SY5Y细胞的细胞生长速率,抑制了细胞活力并减少了集落形成。流式细胞术分析显示,在用shRNA-herg1或shRNA-herg1/1b沉默herg1或herg1/1b基因后,SH-SY5Y细胞在G0-G1期受到阻滞。在体内,瘤内注射shRNA-herg1/1b抑制了裸鼠皮下接种的SH-SY5Y肿瘤的生长。结果表明,herg在调节SH-SY5Y细胞的生长和增殖中起重要作用。阻断HERG通道可能是神经母细胞瘤和其他一些herg基因过表达肿瘤的潜在治疗策略。