磷酸酶基因PTEN对骨肉瘤细胞凋亡机制研究
投稿时间:2014-12-02  修订日期:2015-01-07  点此下载全文
引用本文:张昊,王勇,成志勇,张金金,鲁会亮,张楠.磷酸酶基因PTEN对骨肉瘤细胞凋亡机制研究[J].医学研究杂志,2015,44(8):53-55
DOI: 10.11969/j.issn.1673-548X.2015.08.016
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作者单位E-mail
张昊 671000 大理学院基础医学院人体解剖学教研室
071000 保定市骨科医院骨科 
 
王勇 671000 大理学院基础医学院人体解剖学教研室 347754029@qq.com 
成志勇 071000 保定市第一医院血液内科  
张金金 071000 保定市第一医院血液内科  
鲁会亮 071000 保定市骨科医院骨科  
张楠 071000 保定市第一医院血液内科  
基金项目:云南省科技计划项目(12031586)
中文摘要:目的 探讨肿瘤抑制基因PTEN对人骨肉瘤细胞系MG-63细胞凋亡的影响及其分子作用机制。 方法 将携带有野生型PTEN及绿色荧光蛋白的腺病毒(Ad-PTEN-GFP)及空载体腺病毒(Ad-GFP),转染MG-63细胞系,流式细胞仪分析转染效率及细胞凋亡率,半定量RT-PCR检测PTEN mRNA水平变化,Western blot法检测PTEN蛋白表达,试剂盒检测caspase-3、caspase-7、caspase-9蛋白活性。 结果 以感染复数为100,转染MG-63细胞2天后腺病毒感染效率即达93.2%±4.7%。转染PTEN基因5天后,细胞凋亡率为29.8%,细胞形态出现明显凋亡改变。分子学检测结果显示转染PTEN基因后,PTEN mRNA及蛋白表达明显升高;caspase-3、caspase-7、caspase-9蛋白活性明显增加。 结论 PTEN基因可能通过提高caspase-3、caspase-7、caspase-9活性,诱导人骨肉瘤MG-63细胞凋亡。
中文关键词:PTEN基因  MG-63细胞系  caspase家族  骨肉瘤
 
Research of PTEN Gene on Apoptosis in Osteosarcoma Cells
Abstract:Objective To investigate the mechanism of tumor-suppressing gene PTEN on the cell apoptosis and the influence of Caspase family on human osteosarcoma cell line MG-63 cells in vitro. Methods The Recombinated adenovirus which contained PTEN gene(Ad-PTEN-GFP) and the empty vector (Ad-GFP) were transfected into MG-63 cells respectively. The transfection efficiency of Ad-PTEN-GFP and apoptosis rate were assessed by flow cytometry (FCM). The apoptosis was detected by morphological characteristics of light microscope.The PTEN mRNA levels were detected by reverse transcriptional PCR. Western blot was used to detected PTEN protein. Caspase-3,caspase-7 and caspase-9 protein activities were tested by kits. Results When multiplicity of infection (MOI) was 100, the efficiency of denovirus transfection MG-63 cell was 93.2%±4.7% after transfection 48h. The apoptosis ratio was 29.8% after MG-63 cells transfected with Ad-PTEN-GFP 5 days. The apoptosis appearance with morphological characteristics appeared, PTEN mRNA and protein increased, the caspase-3,caspase-7,caspase-9 protein activity was up-regulated respectively compared with Ad-GFP group after transfaction of PTEN gene. Conclusion PTEN gene can induce MG-63 cells apoptosis probability via up-regulating the caspase-3,caspase-7,caspase-9 ability.
keywords:PTEN gene  MG-63 cell line  Caspase family  Osteosarcoma
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