三氧化二砷诱导小鼠小脑组织细胞凋亡相关基因改变的芯片研究

Changes of apoptosis-associated genes expression in cerebellar cells of mice exposed to Arsenic trioxide

关键词:  As2O3  小脑组织  细胞凋亡  基因芯片  差异表达

Key words:  As₂O₃  cerebellar cells  apoptosis  genechip  differential expression of gene

摘要:

［目的］利用基因芯片研究As₂O₃ 诱导小鼠小脑组织细胞凋亡信号转导相关基因表达的差异性，寻找关键基因，并分析其可能的作用机制。 ［方法］小鼠通过自然饮用含As₂O₃ 自来水方式进行砷暴露60 d后，对其小脑组织进行总RNA提取、纯化、体外转录合成cRNA探针、cRNA探针生物素标记、与Mouse Genome 430 2.0 Araay基因芯片杂交、扫描杂交信号、最后进行芯片数据处理和生物学信息分析。［结果］与对照组比较，Mdfic、Pycard、Igh-6在染毒组表达上调，Igf1r、Mapk8ip1、Prdx2、Frag8、Spred2在染毒组表达下调，但其在低剂量组与高剂量组表达水平相同；与对照组比较， Arhgdig、Aldh1a1、Nudt2、Il18在染毒组表达上调，Mapk8、Bcl2l1表达下调，且差异表达程度呈剂量反应关系；与对照组和低剂量组比较，高剂量组的Cartpt、Map4k2表达上调，Rock1、Cacna1α、Hipks、Sod1、Arhgap5、Srgap2表达下调。［结论］As₂O₃ 诱导小脑组织细胞凋亡的可能机制主要与Caspase途径及JNK途径的活化和细胞内钙离子浓度的增高有关，所涉及的主要相关基因有：Rock1、Pycard、Cacna1α、Cartpt和Mdfic。

Abstract:

［Objective］To screen critical apoptosis-associated genes in cerebellar cells of mice exposed to arsenic trioxide.［Methods］After 60 days of expousure to arsenic trioxide, total RNA in cerebellar cells of mice was extracted,purified, and then transcribed to cRNA probe. Then, cRNA probe was biotin labeled and hybridized with Mouse Genome 430 2.0 Araay. Hybridization signals were scaned and genechip data was analyzed.［Results］Compared with control group, Mdfic,Pycard,Igh-6 were up-regulated and Igf1r,Mapk8ip1,Prdx2,Frag8,Spred2 were down-regulated in the groups exposed to arsenic trioxide, while the levels of these genes expressions were the same between the two experimental groups. Compared with control group, Arhgdig,Aldh1a1,Nudt2,Il18 were up-regulated and Mapk8,Bcl2l1 were down-regulated in the groups exposed to arsenic trioxide, and the expression levels of these genes show dose response relation. Compared with control group and low-dose group, Cartpt,Map4k2 were up-regulated and Rock1,Cacna1α,Hipks,Sod1,Arhgap5,Srgap2 were down-regulated in high-dose group.［Conclusions］The mechanisms of arsenic-inducing apoptosis in mice cerebellar cells may be mainly associated with the activation of Caspase, JNK signal pathway and the increase of Ca²⁺ concentration. The involved critical genes may be Rock1,Pycard,Cacna1α,Cartp and Mdfic.