大肠癌细胞与血管内皮细胞间缝隙连接通讯的建立与功能检测

摘要

目的 建立大肠癌细胞与血管内皮细胞间缝隙连接细胞通讯(gap junction intercellular communication,GJIC)模型,探讨癌细胞与血管内皮细胞之间GJIC的变化在癌转移中的作用.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及与不同转移能力的人大肠癌细胞系LoVo细胞、HT29细胞共培养之内皮细胞间缝隙连接细胞间通讯的状况.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[(25.68±15.74)%和(15.13±7.76)%,P＜0.05].结论 大肠癌细胞与血管内皮细胞黏附后,其间的缝隙连接细胞间通讯减少,并且高转移能力的LoVo细胞与血管内皮细胞间的缝隙连接通讯减少尤其明显.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发生改变,与恶性肿瘤的转移特性密切相关.%Purpose To establish a gap junction intercellular communication( GJIC )model after colorectal carcinoma cell adhered to vascular endothelial cell ( EC ) and to analyze the relationship between the expression of GJIC and cancer metastasis. Methods Cell culture,immunohistochemistry,fluorescence recovery after photobleaching assay were used to detect the expression of GJIC after ECs were solely cultivated and together with HT29 cells and LoVo cells, respectively. Results In the group of ECs, after the cells were bleached.intercellular fluorescence intensity started to go up , whereas in the group of EC that we cultivated it together with HT29 cell fluorescence recovered slowly,and much more slowly with LoVo cell [ ( 25. 68 ± 15. 74 )％ vs ( 15. 13 ±7. 76 )％ ,P ＜ 0. 05]. Conclusion After colorectal carcinoma cells adhere to EC , GJIC hetween EC and colorectal carcinoma cell decrease,particularly in cocultured with LoVo cells. Expression of GJIC between cancer cells and vascular endothelial cells is closely related to the metastasis of cancer.