转录组分析揭示妊娠期镉暴露对胎盘滋养层细胞活力的影响机制

Transcriptome analysis revealed the effects of cadmium exposure on the activity of placental trophoblast cells during pregnancy

目的探讨妊娠期不同浓度镉(Cadmium,Cd)处理对胎盘滋养层细胞活力存在影响及其相应的分子机制。方法在胎盘滋养层细胞系JEG3细胞中,采用5、10和20μmol/L CdCl_(2)处理24 h后,采用CCK⁃8试剂盒检测细胞活力,同时,在相同条件下应用转录组测序检测不同浓度CdCl_(2)处理对JEG3细胞基因表达及其相关信号通路的影响。结果(1)CdCl_(2)处理会抑制胎盘滋养层细胞活力,10和20μmol/L CdCl_(2)处理会明显抑制JEG3细胞的活力。(2)对5、10和20μmol/L CdCl_(2)处理组与对照组样品进行转录组测序共获得88.6 Gb的质控数据,且任意2个样品间的相关性高于0.878。(3)测序结果显示5、10和20μmol/L CdCl_(2)处理组的明显差异表达基因(DEG)分别为72、476和825个,在3个处理浓度下,金属硫蛋白(MT)家族基因的变化最为明显。(4)3个不同浓度CdCl_(2)处理组中,GO富集分析的差异基因主要集中于细胞过程、单体过程、生物学调控、细胞组分、细胞粘附和催化活性等。(5)KEGG富集分析发现,5与10μmol/L CdCl_(2)处理下矿物质吸收通路富集最为明显,20μmol/L CdCl_(2)处理下AGE⁃RAGE信号通路富集较为明显。结论Cd会抑制滋养层细胞的活力,其主要的机制可能为Cd处理会促进MT家族基因的表达进而通过矿物质吸收通路影响细胞活力,而高浓度(20μmol/L)处理时,Cd不仅通过矿物质吸收通路影响细胞活力,还会通过AGE⁃RAGE信号通路抑制细胞活力。本研究为今后确定Cd暴露导致的胎盘发育与功能异常奠定重要的分子基础。

Objective To investigate the effect of different concentrations of cadmium on the activity of placental trophoblast cells during pregnancy and its corresponding molecular mechanism.Methods Placental trophoblastic JEG3 cells were treated with 5,10 and 20μmol/L CdCl_(2)for 24 hours,then CCK⁃8 kit was used to detect cell viability.Meanwhile,transcriptome sequencing was used to detect the effects of different concentrations of CdCl_(2)on gene expression and related signaling pathways in JEG3 cells under the same conditions.Results(1)The activity of placental trophoblast cells was inhibited by cadmium treatment,and the activity of JEG3 cells was significantly inhibited by 10 and 20μmol/L CdCl_(2)treatment.(2)Transcriptome sequencing of 5,10 and 20μmol/L CdCl_(2)treated groups and control groups obtained a total of 88.6 Gb of clean data,and the correlation between any two samples was higher than 0.878.(3)The sequencing result showed that there were 72,476 and 825 significant differenced genes treated with CdCl_(2)at 5,10 and 20μmol/L,respectively.The changes of MT family genes were the most significant at the three treatment concentrations.(4)In the three CdCl_(2)treatment groups with different concentrations,the differential genes in GO enrichment analysis were mainly concentrated in cell processes,monomer processes,biological regulation,cell components,cell adhesion,catalytic activity,etc.(5)KEGG enrichment analysis showed that the enrichment of mineral absorption pathway was most significant under 5 and 10μmol/L CdCl_(2)treatment,and the enrichment of AGE⁃RAGE signaling pathway was more significant under 20μmol/L CdCl_(2)treatment.Conclusion The cadmium can inhibit the activity of trophoblast cells,the main mechanism may be that cadmium can promote the expression of MT family genes and then affect cell vitality through mineral absorption pathway,while high concentration(20μmol/L)can not only affect cell vitality through mineral absorption pathway,but also inhibit cell vitality through AGE⁃RAGE signaling pathway.This study will lay an important molecular foundation for the future identification of placental developmental and functional abnormalities caused by cadmium exposure.