摘要
目的:观察早期生长反应基因1(Egr1)对成骨肉瘤细胞Saos-2中stathmin基因表达调控的影响.方法:构建Egr真核表达载体,应用脂质体转染人成骨肉瘤Saos-2细胞,采用RT-PCR检测转染细胞的Egr1及stathmin mRNA表达的变化Western Blot和免疫细胞化学染色检测stathmin蛋白表达;MT法检测转染细胞生长状况;原位末端标记(TUNEL)检测细胞凋亡情况.结果:RT-PCR检测发现Egr1表达水平明显高于空白对照组,stathmin mRNA表达水平显著降低,转染细胞stathmin蛋白表达水平明显降低.MTT法结果显示Egr1可显著抑制Saos-2细胞的生长,TUNEL阳性细胞明显增加.结论转录调控因子Egr1参与stathmin基因转录调控并可抑制其表达,进而促进Saos-2细胞凋亡.
AIM：To investigate the effects of stathmin gene expressionin osteosarcoma Saos-2 cells induced by early growth responsegene-1（Egr1）.METHODS：To construct pcDNA3.1-Egr1 recombinant plasmid and then exogenous Egr1 gene was introduced into Saos-2 with Lipofectamine transfection method.The expression of Egr1 gene and stathmin gene were detected by RT-PCR,Western Blot and immunocytochemistry.The apoptosis and proliferation of Saos-2 cells were observed by TUNEL and MTT assay.RESULTS：The results of RT-PCR indicated that the expression of Egr1 gene was upregulated,however the expression of stathmin gene was downregulated in the transfected cells.MTT assay revealedthat the proliferation of Saos-2 cells transfected EGR-1 gene was much lower than that of untransfected cells.The results of TUNEL showed that EGR-1 gene induced apoptosis of the transfected cells.CONCLUSION：Egr1 can suppress the expression of stathmin gene and induce Osteosarcoma Saos-2 cell apoptosis.
出处
《第四军医大学学报》
北大核心
2009年第22期2493-2496,共4页
Journal of the Fourth Military Medical University
基金
国家自然科学基金(30772181)
