Gluc-Fluc双荧光素酶质粒转染MB49细胞后荧光表达特性  

Expression of Gluc-Fluc dual luciferase plasmid after transfection into MB49 bladder cells

作　　者：张信基[1] 石小军[1] 孙鹏宇[1] 张哲欢[1] 付昕阳[1] 谭万龙[1] 

机构地区：[1]南方医科大学南方医院泌尿外科,广东广州510515

出　　处：《南方医科大学学报》2011年第3期499-503,共5页Journal of Southern Medical University

基　　金：广东省自然科学基金(9151051501000030)

摘　　要：目的利用生物发光成像技术对Gluc-Fluc双荧光素酶质粒转染MB49膀胱癌细胞后双荧光表达量变化特性进行研究。方法用pAAV2neo-Gluc和pAAV2neoCAG-Gluc-2A-Fluc分别转染MB49细胞,利用荧光照度计来研究双荧光素酶质粒中Gluc、Fluc的特性;利用体外活体成像系统对双荧光素酶质粒Gluc-Fluc转染MB49膀胱癌细胞后荧光表达情况的检测。结果荧光照度计检测结果显示:Gluc随着细胞数的增多或时间的延长,荧光表达量呈相应的增加。Fluc随细胞数目的增多,荧光表达量增加;但随时间的延长,荧光表达量变化不大。体外活体成像结果显示:Gluc-Fluc双荧光素酶质粒已转染入MB49膀胱癌细胞,经过G418加压筛选获得稳定表达的细胞株。结论双荧光素酶基因的构建并未改变Gluc的自身荧光表达特性。转染双荧光素酶质粒的MB49膀胱癌细胞,由于其Fluc在活体成像时的定位优势和Gluc易分泌、检测灵敏度高的定量优势,为后期动态检测肿瘤生长变化以及评价药物治疗效果提供了一个可靠的基石和平台。Objective To investigate the expression characteristics of Gluc-Fluc dual luciferase plasmid after its transfection into MB49 bladder cells.Methods pAAV2neoCAG-Gluc-2A-Fluc and pAAV2neo-Gluc plasmids were separately transfected into MB49 cells via LipofectamineTM2000.The Gluc activity in the cell culture supernatant and the Fluc activity in the cells were detected by luminometer and Lumina Imaging system.Results The luminometer result showed that the activity of Gluc in the supernatant increased gradually in a cell number-and time-dependent manner,while Fluc activity in the cells increased with the cell number but not with time.The Lumina Imaging system showed that Gluc-Fluc was successfully expressed in MB49 bladder cells and cell lines with stable Gluc-Fluc expression were obtained after G418 selection.Conclusion Gluc in the dual luciferase plasmid retains its expression characteristics.Due to the advantages of Fluc in localization in living imaging and the easy quantitative detection of Gluc,the dual luciferase plasmid,after transfection in MB49 bladder cells,allows reliable and dynamic detection of tumor growth in animal models.

关 键 词：双荧光素酶 转染 活体成像 膀胱癌 

分 类 号：R737.14[医药卫生—肿瘤]