胃癌SP表型细胞侵袭转移相关性研究  

The study of invasive ability and lung metastasis of side population phenotypes in human gastric cancer cell line

作　　者：孙力超[1] 彭亮[2] 韩璐璐[1] 李闯[1] 杨治华[1] 冉宇靓[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤研究所分子肿瘤学国家重点实验室,北京100021 [2]中日友好医院临床医学研究所,北京100029

出　　处：《现代肿瘤医学》2011年第5期837-842,共6页Journal of Modern Oncology

基　　金：国家重点基础研究发展计划(973)资助项目(No:2009CB521804)

摘　　要：目的:观察人胃癌SNU5肿瘤细胞系及其高侵袭细胞亚系中是否含有SP细胞,并验证该表型细胞与侵袭转移的关系。方法:采用Transwell小室,体外筛选出高侵袭细胞;经Hoechst33342染色,应用流式检测并分选出SP表型以及非SP表型细胞;应用无血清培养方法检测SNU5亲本细胞、SP表型以及非SP表型细胞在无血清培养中的成球能力。将分选出的SP表型以及非SP表型细胞进行侵袭实验、划痕迁移实验。裸鼠皮下接种,检测SP表型以及非SP表型细胞的自发性肺转移能力。基因芯片的方法分析SP表型以及非SP表型细胞的基因表达谱差异。结果:经过三轮筛选建立了稳定的高侵袭亚细胞群SNU5-P3。SP分选检测发现,SNU5-P3的SP细胞比例为1.6%;无血清培养发现,SNU5-P3-SP细胞成球数为104±19,显著强于SNU5以及SNU5-P3-non-SP细胞。侵袭实验检测发现,SNU5-P3-SP细胞的侵袭能力比SNU5、SNU5-P3-non-SP细胞增强近2.5倍。划痕迁移检测发现,SNU5为-P3-SP运动能力显著增强,划痕24h愈合。体内移植瘤转移实验发现SNU5-P3-SP细胞自发性肺转移率为100%,SNU5为50%,而SNU5-P3-non-SP仅有16.7%。基因芯片分析,获得733个差异基因,其中上调基因450个,36%的基因与侵袭转移有关。结论:胃癌SP表型细胞具有干细胞的特征,在胃癌侵袭、转移中起到关键作用。Objective：To explore side population（SP）phenotypes in gastric cancer cell line and high invasive sub-cell lines and to observe the relationship between SP phenotype and invasion coupled with spontaneous lung metastasis.Methods：High invasive cells were selected from gastric cancer cell line SNU5 using Transwell chamber in vitro.Cancer cells suspension was stained with Hoechst33342 and propidiumiodide（PI）with absence or presence of verapamil.Then SP was analyzed by fluorescence activated cell sorter.The ability of sphere formation,migration and invasion was tested by serum-free culture method,wound healing assay and Transwell assay.We also inoculated with SP phenotype and non-SP cell phenotype of spontaneous to detect the future lung metastasis.Gene chip was applied to analyze the different gene expression profile of SP and non-SP phenotype cells.Results：After three rounds of screening we established a highly invasive subcellular SNU5-P3.SP cells accounted for 1.6%of the SNU5-P3 cells.SNU5-P3-SP cells were cultured in serum-free medium and formed 104±19 sphere cells formation,which was more than sphere cells formed by SNU5-P3 or SNU5-P3-non-SP.The invasive assay showed that the invasive ability of SNU5-P3-SP had 2.5 times of than SNU5-P3 or SNU5-P3-non-SP cells.The wound healing assay showed increased migration capacity of SNU5-P3-SP compared with SNU5-P3 or SNU5-P3-non-SP cells.The ability of metastasis of three cells showed that SNU5-P3-SP had obvious lung metastasis.The visible lung metastasis of SNU5-P3-SP was 100%,SNU5 was 50%and SNU5-P3-non-SP was only 16.7%.Microarray analysis showed that we obtained 733 different genes,of which 450 genes were up-regulated and 36%of them were invasion or metastasis related genes.Conclusion：SP phenotype cells had stem cell characteristics and play roles in the invasion of gastric cancer and lung metastases.

关 键 词：胃癌 SP细胞 侵袭 肺转移 

分 类 号：R73-35[医药卫生—肿瘤] R735.2[医药卫生—临床医学]