论文部分内容阅读
目的通过观察辐射对小鼠皮肤成纤维细胞系L929生长及CCN1基因表达的影响,探讨CCN1基因与辐射损伤的关系。方法体外培养L929细胞系,以4Gy60Coγ射线照射为辐射组,并设对照组,MTT法测细胞增殖变化,平板克隆形成实验检测克隆形成能力,流式细胞术测细胞周期,RT-PCR及免疫细胞化学法检测L929细胞中CCN1基因的表达情况。结果辐射组细胞辐照2d增殖抑制明显(P<0.01),克隆形成也明显受到抑制(P<0.05);辐射后24h,细胞出现G2期停滞;辐射后6hL929细胞CCN1的mRNA表达水平明显降低(P<0.01),至24h开始回调(P<0.05);辐照后24～48h,细胞CCN1蛋白的表达水平明显下降(P<0.01)。结论因辐照生长受到抑制的小鼠成纤维细胞L929,其CCN1基因的表达水平下调,提示CCN1基因的表达水平低下是辐射导致细胞生长抑制的因素之一。
Objective To observe the effect of radiation on the growth of mouse dermal fibroblast cell line L929 and the expression of CCN1 gene and to explore the relationship between CCN1 gene and radiation injury. Methods L929 cell line was cultured in vitro. The cells were irradiated with 4Gy60Coγ ray and the control group was established. The cell proliferation was measured by MTT assay. The clonogenic capacity was assayed by plate clone formation assay. The cell cycle, RT-PCR and immune cells Chemiluminescence was used to detect the expression of CCN1 gene in L929 cells. Results Radiation group showed significant inhibition of proliferation (P <0.01) and clonogenic formation (P <0.05) at 24 h after irradiation, and arrested G2 phase at 24 h after radiation. The expression of CCN1 mRNA decreased significantly at 6 h after radiation (P <0.01), and then began to change back to 24h (P <0.05). After 24-48h irradiation, the expression of CCN1 protein decreased significantly (P <0.01). Conclusion The down-regulation of CCN1 gene expression in mouse fibroblast L929 cells with suppressed growth of irradiation suggests that the low expression of CCN1 gene is one of the factors that cause cell growth inhibition by radiation.