基因芯片表达分析4Gyγ射线对小鼠骨髓c-kit阳性细胞影响

Expression changes of genes in c-kit positive cells after 4 Gy γ-ray irradiation： a gene chip analysis

目的 利用基因芯片技术分析经4 Gyγ射线照射后c-kit（CD117,正常表达于干祖细胞表面的细胞因子受体）阳性细胞与代谢过程相关的基因及信号通路表达变化.方法 利用磁珠分选系统分选出骨髓c-kit阳性细胞.实验分为对照组、4 Gy照射组.取1×10^6个c-kit阳性细胞进行照射,照射剂量率为0.99 Gy/min.照射后将细胞培养18h后取出,进行全基因组的高通量基因芯片检测,并进行Gene Ontology聚类分析和Kyoto Encyclopedia of Genes and Genomes信号通路分析.结果 4Gyγ射线照射引起c-kit阳性细胞包括Pld5、Neu2、Bpgm、Alas2、Satl1、Rdh16、Mccc1、Sat2、Smug1、Cml5、Adhfe1、Idh3a、Slc27a5在内的13个基因表达上调3倍以上,包括Acss2、Phgdh、Psat1、Glb1、Gpam、Dus3l、Fuca2、Impdh1、Dlat、Glb1、Enpp4、Prim2、Mettl10、Slc27a2、Dera、Qdpr、Dus1l、Cdyl2、Dhodh、Srr、Spr、Mical2在内的22个基因表达下调3倍以上,这些基因主要参与嘌呤代谢、嘧啶代谢、三羧酸循环等信号通路.结论 4Gyγ射线照射能引起c-kit阳性细胞中与代谢过程相关的许多基因表达发生变化,这些变化的基因有待进一步的实验验证.

Objective To analyze the metabolic process-related genes and pathway variations in the c-kit （CD 117,a cytokine receptor expressed on the surface of hematopoietic stem cells） positive cells after 4 Gy irradiation by using gene chip technology.Methods c-kit positive cells were sorted by microbead separation system.Two groups were involved：control group and 4 Gy irradiation group.A total of 1 ×10^6 c-kit positive cells received 4 Gy irradiation at a dose rate of 0.99 Gy/min.After the two groups were cultured for 18 h,all the cells were collected and detected by gene chip arrays.Data were processed and analyzed via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways.Results After 4 Gy γ-ray irradiation,13 metabolic process-related genes were up-regulated more than 3 times：Pld5,Neu2,Bpgm,Alas2,Satl1,Rdh16,Mccc1,Sat2,Smug1,Cml5,Adhfe1,Idh3a,and Slc27a5.Additionally,22 genes were down-regulated more than 3 times：Acss2,Phgdh,Psat1,Glb1,Gpam,Dus3l,Fuca2,Impdh1,Dlat,Glb1,Enpp4,Prim2,Mettl10,Slc27a2,Dera,Qdpr,Dus1l,Cdyl2,Dhodh,Srr,Spr,and Mical2.These genes were involved in pyrimidine metabolism,purine metabolism,and citrate cycle （TCA cycle） pathways.Conclusion Many variations of genes and pathways in c-kit positive cells existed after 4 Gy irradiation.However,further laboratory technology is required to confirm these variations.