EDA-A1基因突变体对人脐静脉内皮细胞ECV304增殖及细胞周期的影响

发布时间：2018-01-16 11:34

本文关键词：EDA-A1基因突变体对人脐静脉内皮细胞ECV304增殖及细胞周期的影响　出处：《重庆医学》2014年31期 　论文类型：期刊论文

【摘要】：目的研究少汗型外胚层发育不良症EDA-A1基因突变体对人脐静脉内皮细胞(ECV)周期和增殖活性的影响。方法构建EDA-A1基因编码序列(CDS)突变体和野生型的真核表达载体pcDNA3.1(-)-EDA-A1-M/W,转染人脐静脉内皮细胞,逆转录PCR(RT-PCR)扩增EDA-A1基因,蛋白免疫印迹法(Western blot)法检测EDA-A1蛋白表达,MTT法、流式细胞术检测EDA-A1基因突变体对ECV细胞增殖和细胞周期的影响。结果 RT-PCR、Western blot结果显示,野生型组和突变体组ECV细胞表达EDA-A1基因及其蛋白,而空质粒转染组[pcDNA3.1(-)]和空白对照组细胞无表达。与对照组(空质粒租)相比,突变体组ECV细胞增殖活性下降,生长抑制率为45.70%(P0.01),野生型细胞增殖活性无明显改变(P0.05)。突变体组中有更多细胞进入G0/G1期、S期;野生型组S期细胞增多,G2/M期细胞明显减少(P0.05)。结论突变体和野生型EDA-A1基因对ECV细胞增殖和细胞周期有不同的生物学功能。
[Abstract]:Study of hypohidrotic ectodermal dysplasia mutant EDA-A1 gene on human umbilical vein endothelial cells (ECV) of cycle and proliferation. Methods to construct EDA-A1 gene encoding sequence (CDS) eukaryotic expression vector of pcDNA3.1 mutant and wild type (-) -EDA-A1-M/W was transfected into human umbilical vein endothelial cells, reverse transcription PCR (RT-PCR) EDA-A1 gene was amplified by Western blotting (Western blot) expression of EDA-A1 protein was detected by MTT method. The effect of the mutant EDA-A1 gene were detected by flow cytometry on proliferation and cell cycle of ECV cells. The results of RT-PCR and Western blot results showed that the wild type and mutant group group ECV cells expressing EDA-A1 gene and its protein, and empty plasmid transfection group (- [pcDNA3.1)] and control group and control group. The expression of cell (empty plasmid rent) compared to decreased ECV cell proliferation activity mutant, growth inhibition rate was 45.70% (P0.01), wild type Cell proliferation was not significantly changed (P0.05). The mutant group had more cells in G0/G1 phase, S phase; wild type group S phase cells, G2/M phase cells decreased significantly (P0.05). Conclusion the mutant and wild-type EDA-A1 gene function on ECV cell proliferation and cell cycle were different.

【作者单位】：四川省广元市中心医院口腔科;四川大学口腔疾病重点研究实验室;兰州大学生命科学院细胞生物学研究所;兰州大学口腔医学院口腔修复科;
【分类号】：R329.2
【正文快照】： 临床上先天多个或者全部牙齿缺失的病例以少汗型外胚层发育不良症(hypohidrotic ectodermal dysplasia,HED,OMIM305100)、Axenfeld-Rieger综合征(OMIM180500)、Hallermann-Streiff综合征(OMIM234100)最为常见[1]。HED又称为无汗型外胚层发育不良症(anhidrotic ectodermal dys-

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