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目的探讨十溴联苯醚(BDE-209)暴露对人胚胎干细胞系(FY-h ES-10细胞)印记基因表达的影响。方法将FYh ES-10细胞分别暴露于含终浓度为0(溶剂对照,DMSO浓度为1‰)、1、10、100 nmol/L BDE-209的培养基,每24 h换液1次,连续暴露96 h。测定Ki-67阳性率及母源性印记基因H19、KCNK9、DLX5、UBE3A和父源性印记基因SNRPN、MEST、PEG10、GLIS3的表达水平以及基因组DNA总体甲基化水平。结果与溶剂对照组比较,各剂量BDE-209暴露组FY-h ES-10细胞的Ki-67阳性率和基因组总体DNA甲基化水平均较低,差异有统计学意义(P<0.05);且随着BDE-209暴露剂量的升高,而FY-h ES-10细胞的Ki-67阳性率呈下降趋势。各剂量BDE-209暴露组FY-h ES-10细胞内H19、KCNK9、UBE3A、SNRPN、PEG10基因的表达水平均较低,而DLX5、GLIS3基因的表达水平均较高,差异有统计学意义(P<0.05);MEST基因的表达水平均无明显改变。且随着BDE-209暴露剂量的升高,FY-h ES-10细胞H19、KCNK9、PEG10基因的表达水平均呈下降趋势,GLIS3基因的表达水平呈上升趋势。结论 BDE-209有可能通过影响印记基因的表达从而产生胚胎发育毒性。
Objective To investigate the effect of BDE-209 exposure on the expression of imprinted genes in human embryonic stem cell line (FY-h ES-10). Methods FYh ES-10 cells were exposed to medium containing 1 (final concentration of DMSO) and 1, 10, 100 nmol / L BDE-209 respectively, Exposure 96 h. The positive rate of Ki-67 and the expression levels of maternal marker genes H19, KCNK9, DLX5, UBE3A and the parental imprinting genes SNRPN, MEST, PEG10, GLIS3 and the overall methylation level of genomic DNA were determined. Results Compared with the solvent control group, the Ki-67 positive rate and genomic DNA methylation level of FY-h ES-10 cells in each dose of BDE-209 exposed group were significantly lower (P <0.05). And with the increase of BDE-209 exposure dose, the positive rate of Ki-67 in FY-h ES-10 cells showed a decreasing trend. The expression levels of H19, KCNK9, UBE3A, SNRPN and PEG10 in FY-h ES-10 cells were lower than those in BDE-209 exposed groups, while the expression levels of DLX5 and GLIS3 genes were all higher P <0.05). There was no significant difference in the expression of MEST gene. The expression of H19, KCNK9 and PEG10 in FY-h ES-10 cells decreased with the increase of BDE-209 exposure dose, and the expression of GLIS3 increased. Conclusion BDE-209 may have the potential to cause embryonic developmental toxicity by affecting the expression of imprinted genes.