解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (3): 383-387.doi: 10.3969/j.issn.0529-1356.2014.03.016

• 组织学胚胎学发育生物学 • 上一篇    下一篇

Cdc20基因突变对APC min/+小鼠胚胎成纤维细胞生长的影响

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*   

  1. 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
  • 收稿日期:2013-10-21 修回日期:2014-01-24 出版日期:2014-06-06 发布日期:2014-06-06
  • 通讯作者: 王立生 E-mail:wangls168@163.com
  • 基金资助:

    广东省自然科学基金;深圳市科技计划重点项目

Effects of Cdc20 mutation on growth of mouse embryonic fibroblast

FENG Ju-ling1 ZHAO Lei2 XIE Juan1 MO Ming-shu1 GUI Qing-jun1 YOU Yong1 ZHONG Hui1 WANG Li-sheng 3*   

  1. 1. Department of Diagnostics,the University of South China,Hu’nan Hengyang 421001，China; 2. Department of Hepatopancreatobiliary Surgery, the Second Affiliated Hospital of University of South China,Hu’nan Hengyang 421001，China;3. Department of Gastroenterology, the Second Affiliated Hospital of Jinan University, Guangdong Shenzhen 518000，China
  • Received:2013-10-21 Revised:2014-01-24 Online:2014-06-06 Published:2014-06-06
  • Contact: WANG Li-sheng E-mail:wangls168@163.com

摘要：

目的 观察Cdc20 AAA/+APC min/+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Cdc20 AAA/+APC min/+基因型、Cdc20 AAA/+基因型、APC min/+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APC min/+基因型（P＜0.01）。Cdc20 AAA/+APC min/+基因型MEFs克隆形成能力明显增强。Cdc20 AAA/+APC min/+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APC min/+MEFs，大多数为38对。 结论 Cdc20 AAA/+基因突变促进APC min/+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。

关键词: Cdc20;APCmin/+, 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

Abstract:

Objective  Investigation of biological characteristics of Cdc20A AA/+APC min/+mouse embryonic fibroblast(MEFs) indicate the effect of Cdc20AAA/+on growth of mouse embryonic fibroblast and the possible mechanism. Methods MEFs of Cdc20AAA/+APCmin/+, Cdc20 AAA/+ ,APCmin/+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 Cdc20 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.

Key words: Cdc20, APCmin/+, Embryonic fibroblast, Foci formation assay, Karyotype analysis, Chromosome, Instability, Mouse