摘要
目的:通过建立大肠癌细胞与血管内皮细胞间缝隙连接细胞间通讯(GJIC)模型,检测癌细胞与血管内皮细胞间GJIC以及缝隙连接蛋白Cx43表达的情况,分析其与肿瘤转移的关系。方法:采用细胞培养、免疫组化技术及激光漂白后荧光恢复技术,检测单独培养的内皮细胞间以及共培养的人大肠癌细胞系LoVo细胞、HT29细胞与内皮细胞之间GJIC的状况及Cx43的表达。结果:相邻接触的内皮细胞在激光漂白后出现荧光恢复现象,低转移能力的HT29细胞与内皮细胞共培养组荧光恢复明显减缓;高转移能力的LoVo细胞与内皮细胞共培养组荧光恢复更慢。Cx43免疫组化染色显示,单独培养的内皮细胞的细胞膜呈弥漫强阳性着色;HT29细胞与内皮细胞共培养组阳性细胞数量和着色强度明显低于单独内皮细胞培养组;LoVo细胞与内皮细胞共培养组的细胞几乎无Cx43阳性表达。结论:血管内皮细胞与大肠癌细胞粘附后,细胞间的GJIC减少,Cx43的表达降低,且高转移能力的LoVo细胞与血管内皮细胞间的GJIC减少尤其明显,Cx43的表达显著降低甚至缺失。表明肿瘤细胞与血管内皮细胞粘附后,其间的缝隙连接细胞间通讯将发生改变,从而影响恶性肿瘤的转移。
OBJECTIVE：To analyze the relationship between the changes of gap junction intercellular communication （GJIC）,the expression of Connexin 43 and cancer metastasis by establishing the GJIC model after colorectal carcinoma cell adhered to vascular endothelial cell （EC）.METHODS：Cell culture,immunohistochemistry and fluorescence recovery after laser bleaching assay were used to detect the changes of GJIC and the expression of Connexin 43（Cx43） in solely cultured ECs or Ecs co-cultured with HT29 or LoVo cells.RESULTS：In the group of ECs which were cultivated solely,after the cells were bleached,intercellular fluorescence intensity start to go up,whereas in the group of EC which were cultivated it together with HT29 cell,fluorescence recover slowly,and more slowly with LoVo cell.Cells which were cultivated EC solely were strong positive of Connexin43.In the group which were cultivated EC together with HT29 cell,quantities of positive cells reduced and dyeing intention weakened,and in the group which were cultivated EC together with LoVo cell,Connexin43 almost disappeared.CONCLUSIONS：Colorectal cancer cells and ECs adhesion,the GJIC reduction,Cx43 expression decrease,and when LoVo cells adhesion ECs is particularly evident,Cx43 expression is significantly lower or absent.Show that the tumor cell and ECs adhesion,the GJIC will change,thus affecting the metastasis of malignant tumors.
出处
《中华肿瘤防治杂志》
CAS
2010年第14期1066-1069,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
山东省医药卫生科研项目(HZ117)
关键词
缝隙连接细胞间通讯
激光漂白
CX43
肿瘤转移
gap junction intercellular communication
laser bleaching
Connexin 43
metastasis
