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目的研究2,2’,4,4’,5,5’-六氯联苯(2,2’,4,4’,5,5’-hexachlorobiphenyl,PCB153)对人B淋巴母细胞基因表达谱的影响。方法采用0(对照)和25、100、200μmol/L的PCB153分别处理人B淋巴母细胞2、24、48 h,收集细胞后提取总RNA。PCB153处理24 h的RNA样本用Illumina人HT-12 v4全基因组表达谱芯片筛测差异表达基因,并对差异基因进行基因本体论(gene ontology,GO)分类分析。在PCB153处理2、24、48 h样本中用实时定量PCR对候选差异基因进行验证。结果芯片结果显示,各剂量PCB153处理样本中分别发现161、191、1 006个差异基因,三个剂量组重叠差异基因数为15个,其中,上调基因4个、下调基因11个。选取6个差异基因用定量PCR进行验证,发现2个上调基因(CCDC92和TMEM175)及3个下调基因(CCL22、STK38L和GZMK)的表达改变与芯片结果一致。基因CCDC92、CCL22、GZMK和TMEM175等可影响多种淋巴细胞功能,基因STK38L可作用于细胞周期和细胞凋亡等,基因CCL22和MTDH的异常表达则与多种癌症密切相关。结论体外急性染毒PCB153干扰了人B淋巴母细胞某些重要功能基因的表达,为多氯联苯毒性的分子作用机制提供了依据。
Aim To study the gene expression profiles of human B lymphoblasts from 2,2 ’, 4,4’, 5,5’-hexachlorobiphenyl (PCB153) Impact. Methods Human B lymphoblasts were treated with 0 (control) and 25, 100 and 200 μmol / L PCB153 for 2, 24 and 48 h, respectively. Total RNA was extracted after cells were harvested. The 24-hour RNA samples from PCB153 were used to screen differentially expressed genes with Illumina human HT-12 v4 genome-wide expression microarray. Gene ontology (GO) was used to classify the differentially expressed genes. Candidate differential genes were verified by real-time quantitative PCR in the PCBs processed at 22, 24, and 48 h. Results The results of the chip showed that 161,191 and 1006 differential genes were found in the samples treated with different doses of PCB153, and the number of overlapping and differential genes in the three dose groups was 15, of which 4 genes were up-regulated and 11 genes were down-regulated. Six differential genes were selected and verified by quantitative PCR. The results showed that the expression of two up-regulated genes (CCDC92 and TMEM175) and three down-regulated genes (CCL22, STK38L and GZMK) were consistent with the chip results. Genes CCDC92, CCL22, GZMK and TMEM175 can affect a variety of lymphocyte functions, the gene STK38L can act on cell cycle and apoptosis, abnormal expression of genes CCL22 and MTDH are closely related to a variety of cancers. Conclusion In vitro acute exposure to PCB153 interferes with the expression of some important functional genes in human B lymphoblasts and provides a basis for the molecular mechanism of the toxicity of PCBs.