人宫颈癌细胞Hela移植模型肿瘤生长的荧光分析和卡尺测量的比较

目的建立稳定表达绿色荧光蛋白的人宫颈癌细胞系,建立移植瘤模型并比较移植模型肿瘤生长的荧光分析和卡尺测量的优缺点。方法以Lipofectamine 2000介导chickenβ-actin-GFP-NEO转染人宫颈癌细胞Hela,经梯度浓度G418筛选获得稳定表达绿色荧光蛋白的细胞克隆并扩大培养。BALB/cA-nu裸鼠皮下接种1×10^6个发光细胞使其成瘤,利用活体荧光成像系统和游标卡尺观察肿瘤的生长情况。结果获得了稳定表达GFP的人宫颈癌细胞株,将其接种到裸鼠体内可成瘤。活体荧光成像观察发现,1至3周随着肿瘤体积逐渐增大,平均荧光光子数逐渐增加;4周时随着肿瘤出现明显坏死,平均荧光光子数呈现下降趋势,而游标卡尺测量结果显示肿瘤在4至5周仍然不断的增大。结论绿色荧光蛋白能够在人宫颈癌细胞Hela中长期稳定表达,用绿色荧光蛋白标记的人宫颈癌细胞Hela建立的裸鼠肿瘤模型可以为人宫颈癌研究提供理想的实验材料,应用小动物活体成像系统能够客观定量评价活的肿瘤细胞在动物体内的生长情况,而不是肿瘤体积的变化。

The Comparison of Tumor Growth in the Implanted Mouse Model Detected by Optical Imaging System and a Slide Caliper

Objective To establish a human cervical carcinoma cell line with stable expression of green fluorescent protein and the fluorescent-labeled implanted tumor model,and compare the detection methods of tumor growth by optical imaging system and slide caliper.Method Human cervical carcinoma cells were transfected with chicken β-actin-GFP-NEO by Lipofectamine 2000 reagent and screened with G418 to produce the stable GFP-expressing clone.Then the labeled Hela cells were implanted into the subcutaneous tissue of nude mice and the growth of the implanted tumor was observed and growth curve of the labeled Hela cells was analyzed with the whole-body optical imaging system and the slide caliper.Result The stable GFP-expressing Hela cell lines were obtained,which can grow to tumor mass after subcutaneous transplantation.The in vivo fluorescence imaging analysis showed that the fluorescent intensity of tumor mass increased with the increase in tumor volume in nude mice during first to third week.The fluorescent intensity tended to decline with the increasing necrosis at the fourth to fifth week,while the tumor volume was still increasing.Conclusion A GFP-labeled Hela cell line and a mouse tumor model based on the cell line were established,and the animal model provides useful experimental materials for cervical cancer study.The in vivo fluorescence imaging system could be used to quantitatively evaluate the living tumor cell growth in vivo but not the size or volume of the tumor detected by slide caliper.