军事医学2024，Vol.48Issue(5)：362-368,7.DOI:10.7644/j.issn.1674-9960.2024.05.007
组织蛋白酶B基因敲除抑制布鲁菌胞内感染
Deficiency of cathepsin B suppresses Brucella intracellular infection
摘要
Abstract
Objective To investigate the role of cathepsin B(CTSB)inhuman cervical cancer HeLa cellswith Brucella infections.Methods The ctsb knockout(KO)HeLa cell line was constructed using CRISPR/Cas9.The effects of ctsbKO on cell proliferation and survival of intracellular bacteria in case of Brucella infection were detected by CCK-8 assay and intracellular bacteria count while the molecular mechanism of ctsb regulation on Brucella infection was revealed by Western blotting.Results The ctsbKO cell line was constructed.CCK-8 results showed that KO of ctsb had no impact on cell proliferationwith or without Brucella infection.The results of intracellular bacteria count showed a significant decrease in intracellular Brucella four days after infection in the KO cell line.Western blotting assay suggested that the expression levels of autophagy-related proteins were significantly down-regulated in the KO cell line with long-term Brucella infection.Conclusion KO of ctsb inhibits the intracellular survival of Brucella in long-term infections,providing a potential target for the treatment of chronic brucellosis.关键词
布鲁菌/组织蛋白酶B/自噬/CRISPR/Cas9基因编辑Key words
Brucella/cathepsin B/autophagy/CRISPR/Cas9 gene editing分类
医药卫生引用本文复制引用
陆菁菁,许荷玲,张琦,黄芳,李山虎..组织蛋白酶B基因敲除抑制布鲁菌胞内感染[J].军事医学,2024,48(5):362-368,7.基金项目
国家重点研发计划生物安全专项(2022YFC2600702-002) （2022YFC2600702-002）
