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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 To observe the biological characteristics of Cdc20AAA / + APCmin / + mouse embryonic fibroblasts (MEFs) and to explore the effect of Cdc20 gene mutation on the growth of MEFs and its mechanism. Methods The Cdc20AAA / + APCmin / +, Cdc20AAA / +, APCmin / + and WT mouse embryonic fibroblasts were prepared and their growth curves were drawn. The plate clone formation assay was performed to compare the genotypes of various genotypes Changes in growth characteristics of MEFs. The genotypes of MEFs were analyzed for the number of chromosomes and for the detection of mitotic sister chromatid isolation and for the presence of chromosomal instability. Results The growth of Cdc20AAA / + APCmin / + genotype MEFs was faster than that of APCmin / + genotypes (P <0.01). The ability of clonal formation of Cdc20AAA / + APCmin / + genotype MEFs was significantly enhanced. Early segregation of sister chromatids was observed in Cdc20AAA / + APCmin / + MEFs, with an abnormal chromosome number more than APCmin / + MEFs, most of which were 38 pairs. Conclusions The Cdc20AAA / + gene mutation promotes the growth and proliferation of APCmin / + mouse embryonic fibroblasts, which shows the growth characteristics of tumor cells. The mechanism may be related to chromosome instability.