nm23-H1基因对实验性裸鼠肺癌细胞休眠作用的研究  

The Effect of the nm23-H1 Gene in Promoting Lung Cancer Cell Dormancy in Nude Mice

作　　者：王延军[1] 刘伦旭[2] 车国卫[2] 朱文[2] 朱大兴[2] 

机构地区：[1]陕西省延安市人民医院心胸外科 [2]四川大学华西医院胸外科四川省重点实验室华西医院肺癌分子实验室,成都市610041

出　　处：《中国肿瘤临床》2009年第18期1063-1066,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:30400199);四川省应用基础研究计划项目资助(编号:04JY029-084-2)~~

摘　　要：目的:建立人非小细胞肺癌实验性裸鼠肺部的肿瘤细胞休眠模型,并验证nm23-H1基因是否具有促进肿瘤细胞休眠的作用。方法:L9981-nm23-EGFP是nm23-H1基因以真核载体pEGFP质粒转染L9981所构建的人非小细胞肺癌细胞株。L9981-EGFP是转染真核空载体pEGFP质粒的不合nm23-H1基因的人高转移性非小细胞肺癌细胞株。以L9981-nm23-EGFP肺癌细胞作为实验组,空载高转移性肺癌细胞株L9981-EGFP作为对照组,分别接种裸鼠,8周后处死全部裸鼠,检测2组裸鼠肺部肺癌细胞的Ki-67增殖指数,凋亡指数,肿瘤组织VEGF的表达,裸鼠肺组织的调亡指数和G_0/G_1期细胞占细胞总数的百分比。结果:L9981-nm23-EGFP组免疫组化增殖指数、凋亡指数和VEGF的表达均低于高转移性L9981-EGFP组。流式细胞术检测裸鼠肺组织的凋亡指数亦支持免疫组化的结果。实验组G_0/G_1期细胞占细胞总数的比率高于对照组。结论:裸鼠肺部有肺癌休眠细胞存在,实验组裸鼠肺部肺癌休眠细胞多于对照组。nm23-H1基因可能具有促进肺癌细胞休眠的作用。Objective： To establish an animal model for human non-small cell lung cancer （NSCLC） cell dormancy in nude mice and to verify the effect of the nm23-H1 gene on promoting dormancy of NSCLC cells. Methods： L9981-EGFP is a cell line made by transfecting the control eukaryotic expression vector pEGFP into the highly metastatic human NSCLC L9981 cell line, which does not express the nm23-H1 gene. L9981-nm23-EGFP is L9981 transfected with plasmid pEGFP-nm23-H1, a eukaryotic expression vector that contains the nm23-H1 gene. L9981-nm23-EGFP ceils or L9981-EGFP cells were inoculated into nude mice by tail vein injection to establish the NSCLC experimental model. All mice were sacrificed 8 weeks after inoculation. Immunohistochemistry （SP） and TUNEL techniques were employed to measure the proliferation index （PI） using Ki-67, the expression of VEGF in lung cancer tissues and the apoptosis index （Al） of lung cancer tissues. The determination of apoptosis index （Al） and percentage of G0/G1 cells in nude mice lung tissues were also carried out by flow cytometry （FCM）. Results： Immunohistochemistry showed that PI, AI and expression intensity of VEGF in the L9981-nm23-EGFP group were significantly lower than those in the L9981-EGFP group. The AI in nude mice lung tissues as determined by FCM also supported the AI estimations based on immunohistochemistry. The ratio of G0/G1 cells to total cells in the experimental group was significantly higher than that in the control group. Conclusion： There are dormant lung cancer cells in our experimental nude mice lung tissues, significantly more than in the control group. An experimental human NSCLC dormant cell model in nude mice has been successfully established, and it has indicated that the nm23-H1 gene may play a role in the mechanism of lung cancer cell dormancy.

关 键 词：肿瘤休眠 nm23-H1肺癌 实验性动物模型 

分 类 号：R734.2[医药卫生—肿瘤] R735.7[医药卫生—临床医学]