Molecular signatures of cytotoxic effects in human embryonic kidney 293 cells treated with single and mixture of ochratoxin A and citrinin
Molecular signatures of cytotoxic effects in human embryonic kidney 293 cells treated with single and mixture of ochratoxin A and citrinin
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赭曲霉毒素 A 和柑橘素单一和混合物处理人胚肾 293 细胞细胞毒性作用的分子特征
DOI：
10.1016/j.fct.2018.11.015
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发表时间：
2019-01-01
影响因子：
4.3
通讯作者：
Jiang, Yueming
中科院分区：
文献类型：
--
作者：
Gong, Liang;Zhu, Hong;Jiang, Yueming
赭曲霉毒素A(OTA)和桔霉素(CTN)是重要的真菌毒素，常同时存在于食品和饲料中。本研究通过四甲基偶氮唑盐比色法检测OTA和CTN对人胚胎肾293细胞的单独和联合杀伤作用，发现OTA和CTN联合作用对人胚胎肾293细胞有明显的协同杀伤作用，表现为细胞明显积聚于S和G2/M期。转录本和sRNA测序用来探索介导单个或联合细胞毒性的分子特征。共鉴定出378个miRNAs，其中针对数千个基因的66个miRNAs在不同处理下差异表达，120个差异表达基因(Deg)受到单独或联合处理的调控。用荧光素酶检测系统分析了两个具有代表性的miRNAs(hsa-miR-1-3p和hsa-miR-122-5p)及其靶基因细胞程序性死亡10(PDCD10)和细胞周期蛋白G1(CCNG1)与细胞凋亡信号和细胞周期的关系。进一步，通过实时定量聚合酶链式反应和免疫印迹分析验证了它们的表达模式，表明miRNA-靶相互作用可能是霉菌毒素诱导的细胞死亡的原因。综上所述，这些发现为OTA和CTN单独或混合暴露于HEK293细胞的协同细胞毒作用提供了分子证据。
Ochratoxin A (OTA) and citrinin (CTN) are important mycotoxins, which often coexist in food and feed stuff. In this study, individual and combinative cytotoxicity of OTA and CTN were tested in human embryonic kidney (HEK) 293 cells via MTT assay, and synergistic cytotoxic effects were found following co-treatment with OTA and CTN, manifested by significant accumulation of HEK293 cells in S and G2/M stages. Transcriptomic and sRNA sequencing were performed to explore molecular signatures mediating individual or combinative cytotoxicity. A total of 378 miRNAs were identified, among which 66 miRNAs targeting thousands of genes were differentially expressed in response to different treatments, and 120 differentially expressed genes (DEGs) were regulated by either individual or combinative treatments. Correlations between two representative miRNAs (hsa-miR-1-3p and hsa-miR-122-5p), and their target genes, programmed cell death 10 (PDCD10) and cyclin G1 (CCNG1), associated with apoptotic signaling and cell cycle were analyzed by luciferase assay system. Further, their expression patterns were validated by quantitative real-time PCR and western blot analysis, suggesting that both miRNA-target interactions might account for the mycotoxin-induced cell death. Taken together, these findings provide molecular evidences for synergistic cytotoxic effects of exposure to single and mixture of OTA and CTN in HEK293 cells.