BDE-209/BPA暴露对人胚胎干细胞系FY-hES-10早期神经分化中印记基因表达的影响
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摘要
目的探讨十溴联苯醚(BDE-209)和双酚A(BPA)暴露对人胚胎干细胞系FY-hES-10体外早期神经分化中印记基因表达的影响。方法利用添加小分子抑制剂的方法将FY-hES-10细胞系进行神经贴壁诱导,并在培养基中添加低剂量的BDE-209或/和BPA,每24 h换液1次,连续暴露11 d。收集诱导分化第11天的细胞检测nestin阳性率以及印记基因SNRPN、KCNK9、UBE3A和PEG10的表达水平。结果 BDE-209、BPA单独或联合暴露组nestin的阳性率明显低于对照组(P<0. 05)。印记基因SNRPN、KCNK9、UBE3A在BDE-209、BPA单独或联合暴露组表达均明显低于对照组,PEG10在BDE-209 1 nmol/L和BDE-209 1 nmol/L+BPA 1 nmol/L组表达明显低于对照组,而在BPA 1 nmol/L组中表达与对照组无明显区别。结论低剂量BDE-209/BPA单独或联合暴露均有可能通过影响胚胎干细胞早期神经分化中印记基因表达从而产生神经发育毒性,BDE-209和BPA联合暴露可能加重神经发育毒性。
Objective To study the effects of decabrominated biphenyl ether( BDE-209) and/or bisphenol A( BPA) exposure on imprinting gene expression in early neural differentiation of human embryonic stem cell line FY-hES-10. Methods Neuroprogenitor cells were derived from human embryonic stem cell line FY-hES-10 by using small-molecule inhibitors and were exposed to BDE-209 and/or BPA for 11 days. Nestin and imprinting genes( SNRPN,KCNK9,UBE3 A and PEG10) expressions were detected by immunofluorescence staining and q-PCR. Results Compared with the solvent control group,the nestin positive rate of FY-hES-10 derived neuro-progenitors cells in BDE-209 and/or BPA exposure groups were lower. The expression levels of SNRPN,KCNK9 and UBE3 A in BDE-209 and/or BPA exposure groups were lower,the expression levels of PEG10 were also lower in BDE-209 1 nmol/L and BDE-209 1 nmol/L +BPA 1 nmol/L groups with statistically significant differences( P < 0. 05). The expression level of PEG10 did not change significantly in BPA 1 nmol/L exposure group. Conclusions BDE-209 and/or BPA may affect imprinting genes expression resulting in neurodevelopmental toxicity.
Objective To study the effects of decabrominated biphenyl ether( BDE-209) and/or bisphenol A( BPA) exposure on imprinting gene expression in early neural differentiation of human embryonic stem cell line FY-hES-10. Methods Neuroprogenitor cells were derived from human embryonic stem cell line FY-hES-10 by using small-molecule inhibitors and were exposed to BDE-209 and/or BPA for 11 days. Nestin and imprinting genes( SNRPN,KCNK9,UBE3 A and PEG10) expressions were detected by immunofluorescence staining and q-PCR. Results Compared with the solvent control group,the nestin positive rate of FY-hES-10 derived neuro-progenitors cells in BDE-209 and/or BPA exposure groups were lower. The expression levels of SNRPN,KCNK9 and UBE3 A in BDE-209 and/or BPA exposure groups were lower,the expression levels of PEG10 were also lower in BDE-209 1 nmol/L and BDE-209 1 nmol/L +BPA 1 nmol/L groups with statistically significant differences( P < 0. 05). The expression level of PEG10 did not change significantly in BPA 1 nmol/L exposure group. Conclusions BDE-209 and/or BPA may affect imprinting genes expression resulting in neurodevelopmental toxicity.
引文
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[3]Elsworth JD,Jentsch JD,Vandevoort CA,et al.Prenatal exposure to bisphenol A impacts midbrain dopamine neurons and hippocampal spine synapses in non-human primates[J].Neurotoxicology,2013,35C:113-120.
[4]邵敏,陈永亨,李晓宇.人体血清中的多溴联苯醚、邻苯二甲酸酯和双酚A的连续在线分离及气相色谱-质谱测定[J].分析化学,2012,40:1139-1146.
[5]Gregg C,Zhang J,Weissbourd B,et al.High-resolution analysis of parent-of-origin allelic expression in the mouse brain[J].Science,2010,329:643-648.
[6]Fan Y,Luo Y,Chen X,et al.A modified culture medium increases blastocyst formation and the efficiency of human embryonic stem cell derivation from poor-quality embryos[J].J Reprod Dev,2010,56:533-539.
[7]Morizane A,Doi D,Kikuchi T,et al.Small-molecule inhibitors of bone morphogenic protein and activin/nodal signals promote highly efficient neural induction from human pluripotent stem cells[J].J Neurosci Res,2011,89:117-126.
[8]Frederiksen M,Thomsen M,Vorkamp K,et al.Patterns and concentration levels of polybrominated diphenyl ethers(PBDEs)in placental tissue of women in Denmark[J].Chemosphere,2009,76:1464-1469.
[9]Xing T,Chen L,Tao Y,et al.Effects of decabrominated diphenyl ether(PBDE 209)exposure at different developmental periods on synaptic plasticity in the dentate gyrus of adult rats in vivo[J].Toxicol Sci,2009,110:401-410.
[10]Balakrishnan B,Henare K,Thorstensen EB,et al.Transfer of bisphenol A across the human placenta[J].Am JObstet Gynecol,2010,202:393.e1-7.
[11]Sonntag KC,Pruszak J,Yoshizaki T,et al.Enhanced yield of neuroepithelial precursors and midbrain-like dopaminergic neurons from human embryonic stem cells using the bone morphogenic protein antagonist noggin[J].Stem Cells,2007,25:411-418.
[12]Barel O,Shalev SA,Ofir R,et al.Maternally inherited Birk Barel mental retardation dysmorphism syndrome caused by a mutation in the genomically imprinted potassium channel KCNK9[J].Am J Hum Genet,2008,83:193-199.
[13]Kishino T,Lalande M,Wagstaff J.UBE3A/E6-AP mutations cause Angelman syndrome[J].Nat Genet,1997,15:70-73.
[14]Ono R,Nakamura K,Inoue K,et al.Deletion of Peg10,an imprinted gene acquired from a retrotransposon,causes early embryonic lethality[J].Nat Genet,2006,38:101-106.
[15]Lee SH,Appleby V,Jeyapalan JN,et al.Variable methylation of the imprinted gene,SNRPN,supports a relationship between intracranial germ cell tumours and neural stem cells[J].J Neurooncol,2011,101:419-428.