Cdc20基因突变对APC^(min/+)小鼠胚胎成纤维细胞生长的影响  

Effects of Cdc20 mutation on growth of mouse embryonic fibroblast

作　　者：冯聚玲[1] 赵磊[2] 谢娟[1] 莫明树[1] 桂庆军[1] 游咏[1] 钟慧[1] 王立生[3] 

机构地区：[1]南华大学诊断学教研室,湖南衡阳421001 [2]南华大学附属第二医院肝胆胰外科,湖南衡阳421001 [3]暨南大学第二临床学院消化内科,广东深圳518000

出　　处：《解剖学报》2014年第3期383-387,共5页Acta Anatomica Sinica

基　　金：广东省自然科学基金资助项目(10151802001000002);深圳市科技计划重点资助项目(201201013)

摘　　要：目的观察Cdc20AAA/+APCmin/+基因型小鼠胚胎成纤维细胞(MEFs)的生物学性状,探讨Cdc20基因突变对MEFs生长的影响及机制。方法制作Cdc20AAA/+APCmin/+基因型、Cdc20AAA/+基因型、APCmin/+基因型和野生型(WT)小鼠胚胎成纤维细胞,绘制生长曲线、进行平板克隆形成实验,比较各种基因型MEFs生长特性的改变。各种基因型MEFs核型分析染色体个数,并检测有丝分裂姐妹染色单体分离情况及是否存在染色体不稳定。结果 Cdc20AAA/+APCmin/+基因型MEFs生长速度快于APCmin/+基因型(P<0.01)。Cdc20AAA/+APCmin/+基因型MEFs克隆形成能力明显增强。Cdc20AAA/+APCmin/+MEFs中可见姐妹染色单体的提前分离,且染色体数目异常要多于APCmin/+MEFs,大多数为38对。结论 Cdc20AAA/+基因突变促进APCmin/+小鼠胚胎成纤维细胞生长及增殖,使其呈现肿瘤细胞的生长特性,其机制可能与染色体不稳定有关。Objective Investigation of biological characteristics of Cdc 20^AAA/＋APC^min/＋ mouse embryonic fibroblast（MEFs） indicate the effect of Cdc20^AAA/＋on growth of mouse embryonic fibroblast and the possible mechanism . Methods MEFs of Cdc20^AAA/＋APC^min/＋, Cdc20^AAA/＋, APC^min/＋ and WT genotype were harvested from embryos for analysis.The growth characteristics of Cdc20^AAA/＋APC^min/＋, Cdc20^AAA/＋,APC^min/＋and WT mouse embryonic fibroblast were analyzed through growth curve analysis and foci formation assay .Separation of sister chromatid and the presence of aneuploid were detected by karyotype analysis .Results Cell proliferation assays showed that Cdc 20^AAA/＋APC^min/＋cells grew at an accelerated rate compared with APC min/＋MEFs（P〈0.01）.Foci formation assay showed that the clone forming ability was significantly increased .Cdc20^AAA/＋APC^min/＋MEFs showed a significant increase in the frequency of aneuploid compared with WT MEFs , which had a karyotype of 38 and contained prematurely separated sister chromatids .Conclusion Cdc20 carrying a null allele （Cdc20^AAA/＋） may accelerate the growth and proliferation of APC min/＋MEFs and present the growth characteristics of the tumor cells .The possible mechanism may be associated with chromosome instability .

关 键 词：Cdc20 APC^min + 胚胎成纤维细胞 平板克隆形成实验 核型分析 染色体 不稳定 小鼠 

分 类 号：R735.35[医药卫生—肿瘤]