GCDH在抗癌药物斑蝥素生物合成机制中的功能探索  

Functional exploration of GCDH in the biosynthesis mechanism of anticancer drug cantharidin

作　　者：廖玉凤[1] 王宇[2] LIAO Yufeng;WANG Yu(Clinical Medical College of Tianjin Medical University,Tianjin 300000,China;Medical School of Panzhihua University,Panzhihua 617000,China)

机构地区：[1]天津医科大学临床医学院,天津300000 [2]攀枝花学院医学院,四川攀枝花617000

出　　处：《医学研究与教育》2025年第1期14-25,共12页Medical Research and Education

基　　金：国家自然科学基金面上项目(30873377)。

摘　　要：目的获取眼斑芫菁戊二酰辅酶A脱氢酶(Glutaryl-CoA dehydrogenase,GCDH)基因全长cDNA序列,并探索GCDH在抗癌药物斑蝥素生物合成机制中的潜在功能。方法从早期构建的眼斑芫菁cDNA文库中筛选出GCDH片段,利用RACE技术扩增该基因全长cDNA序列,通过RT-qPCR测定眼斑芫菁成虫不同时期(5、10、15、22、25、30 d)GCDH的表达量,并利用在线NCBI数据库进行Blast比对,利用在线资源对GCDH进行结构生物学预测。结果本研究成功获取了眼斑芫菁GCDH基因的全长cDNA序列1338 bp,登录号为MK370842。GCDH的开放阅读框为1260 bp,共编码419个氨基酸残基,系统进化分析结果显示,该基因编码的蛋白与赤拟谷盗(XP_971662)在系统进化关系上最近。RT-qPCR结果显示,该基因在眼斑芫菁雌、雄成虫的表达具有差异性,且在雄虫斑蝥素大量合成的前期(15 d)大量表达。结论眼斑芫菁中的GCDH基因在斑蝥素生物合成过程中可能扮演着重要角色。特别是在雄虫中,GCDH在斑蝥素大量合成的前期大量表达,这进一步提示,氨基酸代谢可能与斑蝥素的合成具有密切的相关性。这些发现为进一步研究斑蝥素生物合成机制及人工养殖获取高含量斑蝥素的斑蝥提供可借鉴的思路和理论基础。Objective To obtain the full-length cDNA sequence of Glutaryl-CoA dehydrogenase(GC-DH)gene and explore the potential functions of GCDH in the biosynthesis of cantharidin.Methods GCDH fragments were screened from the early constructed cDNA library,and the full-length cDNA se-quence of the gene was amplified by RACE technique.The expression of GCDH in adults at different periods(5 days,10 days,15 days,22 days,25 days,30 days)was determined with RT-qPCR.Blast comparison was conducted using online NCBI database,and structural biology prediction of GCDH was made using online resources.Results In this study,the full length cDNA sequence of GCDH 1338 bp was obtained,and the entry number was MK370842.The open reading frame of GCDH was 1260 bp and encodes 419 amino acid residues.Phylogenetic analysis results showed that the protein encoded by this gene was the closest phylogenetic relation to Trbolium castaneum(XP_971662).RT-qPCR results showed that the expression of this gene was different in male and female adults,and it was highly ex-pressed in the early stage(15 d)of mass synthesis of cantharidin in males.Conclusion The GCDH in cantharidin may play an important role in cantharidin biosynthesis.Especially in males,GCDH is high-ly expressed in the early stage of cantharidin mass synthesis,which further suggests that amino acid metabolism may be closely related to cantharidin synthesis.These findings provide useful ideas and the-oretical basis for further study of cantharidin biosynthesis mechanism and artificial cultivation of cantharidin with high content.

关 键 词：戊二酰辅酶A脱氢酶 眼斑芫菁 斑蝥素生物合成机制 

分 类 号：Q966[生物学—昆虫学]