基于RNA-seq技术分析2,6-DCBQ对T24细胞的毒性作用机制  

RNA-seq analysis reveals the toxic effects of 2,6-DCBQ on T24 bladder cancer cells

作　　者：陈福彬 刘宁[1] 季佳佳[1] 陈国敏 方道奎[1] CHEN Fu-bin;LIU Ning;JI Jia-jia;CHEN Guo-min;FANG Dao-kui(Shenzhen Center for Disease Control and Prevention,Shenzhen,Guangdong 518055,China;School of Public Health,Sun Yat-sen University,Guangzhou,Guangdong 510080,China)

机构地区：[1]深圳市疾病预防控制中心,广东深圳518055 [2]中山大学公共卫生学院,广东广州510080

出　　处：《毒理学杂志》2025年第1期27-34,共8页Journal of Toxicology

基　　金：广东省医学科研基金(A2022405);深圳市医学重点学科建设经费(SZXK067);深圳市科技计划项目(JCYJ20210324134409026);深圳市“医疗卫生三名工程”项目资助(SZSM202311001)。

摘　　要：目的检测2,6-DCBQ引起的T24细胞毒性效应并探究其毒作用机制。方法将膀胱癌细胞T24暴露在10~250μmol 2,6-DCBQ中,通过CCK-8试剂盒检测细胞活力并计算其IC_(50);在1/4 IC_(50)、1/2 IC_(50)和IC_(50)3个2,6-DCBQ染毒剂量下,通过乳酸脱氢酶细胞毒性检测试剂盒检测2,6-DCBQ对T24的细胞毒性作用,并通过MDA检测试剂盒检测T24细胞氧化应激水平,最后通过转录组测序探究2,6-DCBQ对T24细胞基因表达和相关信号通路的影响。结果2,6-DCBQ以剂量依赖方式引起T24细胞活力降低、细胞死亡率增加。同时,高剂量2,6-DCBQ暴露也引起细胞氧化损伤(与对照组比较,F=8.725,P<0.05)。转录组测序结果提示2,6-DCBQ暴露导致T24细胞线粒体自噬相关基因异常上调,并抑制胰岛素信号通路相关基因表达。结论2,6-DCBQ暴露诱导细胞发生氧化损伤,导致细胞死亡,线粒体自噬相关通路和胰岛素信号通路在其中发挥重要作用。Objective To detect cytotoxic effects induced by 2,6-DCBQ in T24 cells and to elucidate its mechanisms of toxicity.Methods Bladder cancer T24 cells were subjected to 10~250μmol/L 2,6-DCBQ.Cell viability was determined using the CCK-8 assay to calculate the IC_(50).Cytotoxic effects at 1/4 IC_(50),1/2 IC_(50),and IC_(50)concentrations of 2,6-DCBQ were assessed with the lactate dehydrogenase cytotoxicity detection kit.Oxidative stress levels were quantified using the MDA assay.Effects of 2,6-DCBQ on gene expression and signaling pathways in T24 cells were analyzed through RNA-seq.Results Exposure to 2,6-DCBQ resulted in a dose-dependent reduction in T24 cell viability and an increase in mortality.High-dose exposure to 2,6-DCBQ induced oxidative damage(F=8.725,P<0.05).Transcriptome analysis indicated abnormal upregulation of mitochondrial autophagy-related genes and downregulation of genes associated with the insulin signaling pathway.Conclusion Exposure to 2,6-DCBQ induced oxidative damage leading to cell death,with mitochondrial autophagy-related pathways and the insulin signaling pathway identified as significant factors.

关 键 词：消毒副产物 卤代苯醌 2 6-二氯-1 4-苯醌 转录组测序 线粒体自噬 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]