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应用活体内生物发光技术对人高转移大细胞肺癌细胞株l9981成瘤性和转移性研究

期刊名称：中国肺癌杂志,2008;11(3):321-326
时间：0
分类：R734.2[医药卫生—肿瘤;医药卫生—临床医学]
作者机构：[1]四川大学华西医院肺癌分子生物学重点实验室,成都610041, [2]天津市肺癌转移与肿瘤微环境重点实验室,天津医市肺癌研究所,天津医科大学总医院,天津300052
相关基金：本研究受国家自然科学基金重点项目（No.30430300）,天津市科技支撑计划重点项目（No.06YFSZSF05300）和天津市科技支撑计划重大项目（No.07SYSYSF05000）资助
相关项目：nm23基因逆转肺癌转移的分子机理和信号传导调节

关键词：肺肿瘤, 成瘤性, 生物发光, Lung neoplasms Tumorigencity Bioluminescence

中文摘要：

背景与目的应用阳离子脂质体介导的基因转染方法对nm23-H1缺失的人高转移大细胞肺癌细胞株L9981进行荧光素酶基因（Luc）标记，建立稳定高效表达荧光素酶（Luciferase）基因的人高转移大细胞肺癌细胞株L9981-Luc。应用活体生物荧光成像技术，非侵入性地连续检测小鼠（裸鼠和SCID鼠）皮下肿瘤模型发展演进、自发转移的过程。方法将带有荧光素酶基因（Luc）的质粒PGL4．17经阳离子脂质体介导转入人高转移大细胞肺癌细胞株L9981中，筛选出高效稳定表达荧光素酶的细胞株。将转染后的细胞接种于裸鼠和SCID鼠右后腿腹股沟皮下，建立皮下肿瘤模型，利用活体内可见光成像系统观察其在小鼠体内的生长和转移过程。结果转基因人高转移大细胞肺癌细胞株L9981-Luc可在体内、外持续稳定表达荧光素酶，细胞数和发光值成直线相关。成功地建立了表达荧光素酶活性的动物肿瘤皮下自发转移模型。L9981-Luc保留了原有的高转移特性。瘤重和发光强度呈直线相关。结论利用活体内生物发光技术可以非侵袭性地连续示踪肿瘤细胞在活体内的生长和转移过程，为研究肺癌侵袭转移过程及其机理和最佳治疗策略的选择提供了新的手段和工具。

英文摘要：

Background and objective The aim of this work is to study the tumorigenicity and metastasis ability in human large cell lung caner cell line L9981 by in vivo imaging. Methods We firstly transfected the plasmids with firefly luciferase （luc） gene into L9981 cells and then established the stable transfected L9981-luc cell line with G418. Then the positive L9981- Lue cells were implanted subcutaneously into mice and were monitored for tumor growth and micrometastases with in vivo imaging technique. Results The results showed that the bioluminescence density of the stable transfected L9981-Lue cells correlated to the numbers of the tumor cells in vitro. The L9981-Luc cells still keep the high metastasis characterization. After the L9981-Luc cells were implanted into mice subcutaneously for several weeks, we found the metastasis lesions in the different organs of the mice using in vivo imaging machine and the bioluminescence of the tumor correlated with its size. Furthermore, we confirmed the metastasis lesions by scarifying the mice and analyzing with pathological staining. Conclusion We established a stable L9981-Luc cell line with high metastasis character that can be used to analyze the tumor invasion ad metastasis in animal model by in vivo imaging.

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