广东省自然科学基金团队项目(015012),国家教育部骨干教师基金和广东省自然科学基金(000679)资助.
This work was supported by a grant from The National Science Fund for Distinguished Young Scholars of China (69725009).
以绿色荧光蛋白(GFP)作为标记基因转入人类肺癌细胞系(ASTC-a-1),经800 mg/L G418筛选,获得5株高表达细胞系.利用流式细胞仪对GFP表达的稳定性进行了初步研究,结果表明本实验中有些细胞株间GFP表达稳定性有显著差异(P<0.01).将稳定表达的细胞系(3#)植入裸鼠皮下,成瘤后用氩离子激光器产生的488 nm蓝光经扩束后直接激发,瘤体部位发出强烈的绿色荧光.用530 nm长通滤色片滤除激发光,数码相机记录荧光的分布情况.实验尝试利用激光作为激发光源,检测GFP标记的肿瘤细胞在裸鼠中的定位,期望建立一种新的肿瘤早期检测技术并改进肿瘤转移研究的手段,实验取得了阶段性进展.
Human lung cancer cells (ASTC-a-1) were infected with EGFP plasmid vectors, and incubated in RPMI 1640 culture medium supplemented with 15% FCS and 800 mg/L G418. Strong fluorescence of five cell clones were observed after several generations. The flow cytometry was used to determine the GFP expression stability, the results showed that there was significant difference between 2#(3#) and 4#(5#) (P<0.01). The nude mouse, 6 weeks of age, was injected s.c. with a single dose of 3×106 infected tumor cells. The GFP fluorescence was directly exited by 488 nm argon ion laser and recorded by digital camera with 530 nm long pass filter.
金鹰,邢达.肿瘤细胞的标记及其活体荧光成像[J].生物化学与生物物理进展,2002,29(6):942-945
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