PIM1过表达对甲状腺乳头状癌细胞代谢组学的影响及意义  

Impact of PIM1 overexpression on the metabolomics of papillary thyroid carcinoma cells and its significance

作　　者：王佳琦 许茜茜 唐茜 朱欣 WANG Jiaqi;XU Qianqian;TANG Xi;ZHU Xin(Postgraduate Training Base Alliance of Wenzhou Medical University(Zhejiang Cancer Hospital),Hangzhou310022,China;Key Laboratory of Head&Neck Cancer Translational Research of Zhejiang Province,Zhejiang Cancer Hospital,Hangzhou310022,China)

机构地区：[1]温州医科大学研究生培养基地(浙江省肿瘤医院),浙江杭州310022 [2]浙江省头颈肿瘤转化医学研究重点实验室,浙江省肿瘤医院,浙江杭州310022

出　　处：《中国普通外科杂志》2024年第11期1835-1845,共11页China Journal of General Surgery

基　　金：国家自然科学基金资助项目(82072950)。

摘　　要：背景与目的:编码组成型活性丝苏氨酸激酶的致癌基因PIM1表达上调与各种肿瘤的发生和发展有关,笔者前期研究中证实PIM1对甲状腺癌的致癌作用,能够影响甲状腺癌的发生发展及预后。因此,本研究通过代谢组学分析过表达PIM1的甲状腺乳头状癌(PTC)细胞的代谢特性,为进一步研究PIM1如何调节PTC的代谢过程提供依据。方法:构建PIM1过表达载体,建立稳定转染的PTC细胞株(BCPAP);通过qRT-PCR和Western blot验证PIM1过表达的效果;基于液相色谱-质谱联用(LC-MS)技术,对PIM1过表达的BCPAP细胞株(PIM1-OE)及转染空白质粒的对照细胞株(NC)进行代谢组学分析,结合多元统计分析以及KEGG数据库,鉴定并筛选出差异代谢物及代谢通路;基于生物信息学验证PIM1及相关代谢通路与甲状腺癌患者的预后关系。结果:PIM1过表达的BCPAP细胞株中PIM1的mRNA和蛋白表达水平均明显上调(均P<0.05);代谢组学结果显示,PIM1过表达后,细胞内41种代谢物发生变化,其中15种代谢物(L-天冬氨酸、琥珀酸、L-苏氨酸、L-色氨酸、甜菜碱、2-脱氢泛酸、3-吲哚乙腈、D-章鱼碱、吲哚、N-乙酰谷氨酸、肌酸、泛酸、尿毒酸、N-乙酰-L-天冬氨酸、磷酸羟基丙酮酸)差异较为明显(均P<0.05);结合KEGG数据库分析差异代谢产物后发现31条代谢通路存在差异,其中4条代谢通路(丙氨酸、天冬氨酸和谷氨酸代谢,色氨酸代谢,甘氨酸、丝氨酸和苏氨酸代谢,精氨酸和脯氨酸代谢)差异较为明显,且代谢途径的活性均下调(均P<0.05);生物信息学分析结果显示,PIM1的高表达以及甘氨酸、丝氨酸和苏氨酸代谢通路活性下调的甲状腺癌患者的总生存率均明显降低(均P<0.05)。结论:PIM1可能通过调节天冬氨酸、琥珀酸和色氨酸在内的多种氨基酸代谢水平,影响多种代谢途径,改变PTC的代谢状态,进而促进PTC的发生发展过程,其中甘氨酸、丝氨酸和苏氨酸代谢途径�Background and Aims:The oncogene PIM1,encoding a constitutively active serine/threonine kinase,is upregulated in various tumors and closely associated with tumorigenesis and progression.Our previous studies have confirmed the oncogenic role of PIM1 in thyroid cancer,demonstrating its impact on tumor development,progression,and prognosis.This study was conducted to investigate the metabolic characteristics of papillary thyroid carcinoma(PTC)cells overexpressing PIM1 using metabolomics analysis,providing a basis for further exploring how PIM1 regulates metabolic processes in PTC.Methods:A PIM1 overexpression plasmid was constructed,and a stably transfected PTC cell line(BCPAP)was established.The overexpression of PIM1 was verified by qRT-PCR and Western blot.Metabolomics analysis of PIM1-overexpressing BCPAP cells(PIM1-OE)and control cells transfected with an empty vector(NC)was performed using liquid chromatography-mass spectrometry(LC-MS).Differential metabolites and metabolic pathways were identified and screened through multivariate statistical analysis and the KEGG database.Bioinformatics analysis was conducted to explore the relationship between PIM1,associated metabolic pathways,and the prognosis of thyroid cancer patients.Results:The mRNA and protein levels of PIM1 were significantly upregulated in the PIM1-OE BCPAP cells(both P<0.05).Metabolomics analysis identified changes in 41 intracellular metabolites in PIM1-OE cells,with 15 metabolites showing significant differences,including L-aspartic acid,succinic acid,L-threonine,L-tryptophan,betaine,2-dehydropantothenate,3-indoleacetonitrile,D-octopine,indole,N-acetylglutamic acid,creatine,pantothenic acid,uremic acid,N-acetyl-L-aspartic acid,and hydroxyphosphono-pyruvic acid(all P<0.05).KEGG database analysis revealed significant alterations in 31 metabolic pathways,with 4 pathways—alanine,aspartate and glutamate metabolism;tryptophan metabolism;glycine,serine,and threonine metabolism;and arginine and proline metabolism—being notably affected,showing decre

关 键 词：甲状腺肿瘤 甲状腺癌 乳头状 代谢组学 PIM1 

分 类 号：R736.1[医药卫生—肿瘤]