辐射相关基因Necap2基因的克隆、表达及初步功能研究

Cloning, Expression and Function Preliminary Research of Radiation related gene. Necap2

目的寻找辐射相关基因并探索其在辐射损伤中的作用。方法应用表达谱基因芯片技术比较了骨髓造血细胞mRNA在8Gy照射小鼠及健康小鼠之间表达的差异，应用RT—PCR技术从小鼠骨髓细胞中克隆Necap2基因，并应用流式细胞术分析此基因的功能。结果发现并克隆新的辐射相关基因Necap2，其在照射小鼠骨髓细胞中的表达量约是未照射小鼠的10倍。G2／M期阻滞是骨髓造血细胞辐射后一个特征性改变，因此我们探索Necap2对细胞周期的影响。研究发现Necap2基因细胞内表达可引起HEK293细胞G2／M阻滞，s期减少，但对G0／G1期没有明显影响。结论发现一个新的辐射相关基因Necap2，其在辐射过程中可以引起细胞G2／M期阻滞，在此过程中起到重要的调控作用。

Objective To seek new radiation related genes and study its role in radiation induced hematopoietic ceils injury. Methods We have used DNA microassay screening system to compare different expressed genes from mouse bone marrow hematopoietic cells between 8Gy irradiated mice and healthy ones, we cloned the full-length gene with RT-PCR method from mouse bone marrow hematopoietic cells and analyzed the gene function with flow cytometry. Results A new radiation related gene Necap2 was found and cloned, which expression was increased about 10 times compared with the non-irradiated control. G2/M phase arrest was a characteristic change post irradiation in bone marrow cells. So, we next study the influence of Necap2 gene on cell cycle. We found that G2/M phase was arrested, and S phase was decreased in Necap2 gene transfected HEK 293 cells. Conclusions Discovered a new radiation-related genes Necap2, which can cause cell G2 / M phase arrest in the course of radiation and it plays an important regulatory roles in radiation.