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 赵磊 谢娟 莫明树 桂庆军 游咏 钟慧 王立生*

doi:10.3969/j.issn.0529-1356.2014.03.016

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

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*

作者信息 +

Effects of Cdc20 mutation on growth of mouse embryonic fibroblast

FENG Ju-ling¹ ZHAO Lei² XIE Juan¹MO Ming-shu¹ GUI Qing-jun¹ YOU Yong¹ ZHONG Hui¹ WANG Li-sheng ^3*

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文章历史 +

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。

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.

导出引用

冯聚玲赵磊谢娟莫明树桂庆军游咏钟慧王立生*. Cdc20基因突变对APC ^min/+小鼠胚胎成纤维细胞生长的影响[J]. 解剖学报. 2014, 45(3): 383-387 https://doi.org/10.3969/j.issn.0529-1356.2014.03.016

FENG Ju-ling ZHAO Lei XIE Juan MO Ming-shu GUI Qing-jun YOU Yong ZHONG Hui WANG Li-sheng*. Effects of Cdc20 mutation on growth of mouse embryonic fibroblast[J]. Acta Anatomica Sinica. 2014, 45(3): 383-387 https://doi.org/10.3969/j.issn.0529-1356.2014.03.016

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