南方农业学报2023，Vol.54Issue(11)：3349-3358,10.DOI:10.3969/j.issn.2095-1191.2023.11.021
砷制剂对禽白血病肿瘤细胞Warburg效应及Hedgehog信号通路的影响
Effects of arsenic preparations on the Warburg effect and Hedgehog signaling pathway in avian leukemia tumor cells
摘要
Abstract
[Objective]To explore the effects of arsenic trioxide(ATO)on Warburg effect and Hedgehog signaling pathway of avian leukemia(AL)tumor cells,to clarify the mechanism of ATO alleviating AL liver tumor lesions in chick-ens,and to provide scientific basis for ATO treatment of AL in production.[Method]Acute toxicity test was carried out to determine the safety of ATO to green shell laying hens.The pathogen of avian leukemia virus(ALV)was identified by p27 antigen detection,PCR amplification and gp85 gene sequencing,and the pathological diagnosis was made by collect-ing the tumor tissue of infected chickens.AL chicken tumor cells were cultured in vitro and treated with 0.5,1.0 and 2.0 μmol/L ATO,respectively.The cells were collected after culture for 12,24 and 48 h,and the contents of glucose,lactic acid,hexokinase(HK)and glucose transporter 1(GLUT1)were detected,respectively.Green shell laying hens were arti-ficially infected with ALV to construct a model group,and then treated with different doses of ATO(0.5,1.0 and 2.0 mg/kg·BW).ELISA was used to detect the protein levels of Hedgehog signaling pathway Gli1 and Shh in chicken liver tissue.The expression of Shh,Gli1,Gli2,Ptch2 and Smo genes was detected by real-time fluorescence quantitative PCR.[Re-sult]The median lethal dose(LD50)of ATO for green shell laying hens was 19.501 mg/kg·BW,and the LD50-95%confi-dence limit was 19.501±0.213 mg/kg·BW.An ALV-J strain named GZCS01 was identified from green shell laying hens suspected to have AL,and the tumor formed by infection was nephroblastoma.At different doses of ATO(0.5,1.0 and 2.0 μmol/L),ATO was applied to wilms tumor cells.After 48 h culture,glucose content,lactate generation and HK con-tents in cell fluid were extremely significantly decreased(P<0.01,the same below),and GLUT1 content in wilms tu-mor cells was also decreased.Thus,the Warburg effect of tumor cells was weakened.High dose of ATO(2.0 mg/kg·BW)could extremely significantly or significantly inhibit the expression of Shh and Gli1 protein(P<0.05,the same below),extremely significantly or significantly inhibit the expression of Shh,Gli1,Gli2 and Smo genes,and extremely signifi-cantly up-regulate the expression of Ptch2 gene.[Conclusion]ATO can inhibit glucose uptake and lactic acid production of tumor cells,and reduce key enzyme production,thus weakening Warburg effect and inhibiting tumor cell proliferation.ATO can also reduce liver lesions in AL by down-regulating the expression of key factors in Hedgehog signaling pathway such as Shh,Gli1,Gli2 and Smo,and promoting the expression of Ptch2.It can be seen that the use of arsenic prepara-tions(such as arsanic acid)in the treatment of AL is feasible.关键词
三氧化二砷(ATO)/禽白血病(AL)/肿瘤/Warburg效应/Hedgehog信号通路Key words
arsenic trioxide(ATO)/avian leukemia(AL)/tumor/Warburg effect/Hedgehog signaling pathway分类
农业科技引用本文复制引用
任涛,王建俊,杨剑,杨鑫,徐浩,温贵兰,欧德渊..砷制剂对禽白血病肿瘤细胞Warburg效应及Hedgehog信号通路的影响[J].南方农业学报,2023,54(11):3349-3358,10.基金项目
国家自然科学基金项目(32202854) （32202854）
贵州省科技计划项目(黔科合基础[2020]1Y136) （黔科合基础[2020]1Y136）
贵州省科技支撑计划项目(黔科合支撑[2021]一般164号) National Natural Science Foundation of China(32202854) （黔科合支撑[2021]一般164号）
Guizhou Science and Technology Project(QKHJC[2020]1Y136) （QKHJC[2020]1Y136）
Guizhou Science and Technology Support Project(QKHZC[2021]Yiban 164) （QKHZC[2021]Yiban 164）
