摘要
目的研究人垂体瘤转化基因1(hum an p itu itary tumor transform ing gene 1,PTTG1)上调表达对肺腺癌细胞株(A549)增殖和凋亡的影响。方法构建含PTTG1基因的真核表达载体,脂质体法转染A549细胞并用G418筛选出高表达的阳性克隆株,实时荧光定量PCR(real-tim e fluorescent quantitation PCR)和免疫细胞化学(immunocytochem ical assay,ICA)分别检测PTTG1 mRNA及蛋白表达的改变,3H胸腺嘧啶核甙(3H-TdR)掺入法检测细胞增殖活性,AnnexinⅤ异硫氢酸荧光素(FITC)试剂盒检测凋亡情况。结果成功构建含PTTG1基因的真核表达载体,获得上调表达PTTG1的A549细胞,转染含PTTG1基因重组质粒的A549细胞增殖活性较未转染组细胞及转染空白质粒组细胞均显著增强(P<0.05);3组细胞间的凋亡率无显著差异。结论PTTG1基因能显著促进A549细胞的增殖,而对细胞凋亡没有影响。
Objective To investigate the effect of human pituitary tumor transforming gene 1 （ PTTG1 ） transfection on cell proliferation and apoptosis of A549 cells. Methods The eukaryotic expression vector expressing PTTG1 gene was constructed, and transfected into A549 cells by lipofection. The positive clones were screened by using G418. PTTG1 mRNA and PTTG1 protein expression was confirmed by real-time fluorescent quantitation PCR and immunocytochemical assay, the cell proliferation was determined by ^3H-TdR incorporation, and the apoptosis was determined by Annexin V-FITC kit. Results Recombinant expression vector for PTTG1 is successfully constructed and A549 cells highly expressing PTTG1 are obtained. The proliferation of A549 cells transfected with PTTG1 was higher than that of A549 cells and A549 transfected with empty vector （P 〈 0.05 ）, but the apoptosis was of no significant difference among the three groups. Conclusion PTTG1 gene enhances the proliferation and has no effect on the apoptosis in A549 cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2006年第9期961-963,共3页
Journal of Third Military Medical University
关键词
垂体瘤转化基因1
真核表达载体
A549细胞
增殖
凋亡
pituitary tumor transforming gene 1
eukaryotic expression vector
A549 cells
proliferation
apoptosis
