结肠癌细胞起源的白细胞介素1α对血管内皮细胞增殖和迁移的影响  

Effect of colon cancer celi-derived IL-1α on the migration and proliferation of vascular endothelial cells

作　　者：马家驰[1] 陈泉[1] 顾远晖[1] 李一平[1] 房伟[1] 刘美玲[1] 陈晓昌 郭庆金 马世勋[2] 

机构地区：[1]甘肃省人民医院普外科,兰州730000 [2]宁夏医科大学临床学院

出　　处：《中华肿瘤杂志》2015年第11期810-815,共6页Chinese Journal of Oncology

基　　金：基金项目：国家自然科学基金地区科学基金（81260325）;甘肃卫生行业科研计划资助项目（GSWST2010-17）

摘　　要：目的探讨结肠癌细胞起源的白细胞介素1仅（IL—1α）及其受体拮抗剂IL—1α对血管内皮细胞的影响。方法Westernblot法检测IL-1α和IL-1I型受体（IL-1R1）在结肠癌细胞中的表达，细胞增殖和迁移实验检测外源性的IL-1α和IL—1α对胎儿脐静脉内皮细胞（HUVEC）增殖和迁移的影响；采用双层培养系统检测肿瘤细胞起源的IL-1α对血管内皮细胞增殖和迁移的影响，以及IL-1α对血管内皮细胞的作用。结果Westernblot检测显示，IL-1α蛋白在高肝转移结肠癌HT-29和WiDr细胞中表达，而在低肝转移结肠癌CaCo-2和CoL0320细胞中不表达。细胞迁移实验显示，对照组、1ng／mrIL—1α组、10n异／mlrIL—1α组和100ng／mlrIL—1α组的穿膜细胞数分别为（17．9±3．6）个、（23．2±4．2）个、（31．7±4．5）个和（38．6±4．9）个，IL-1α能够显著提高HUVEC的迁移能力，并与IL-1α浓度有关（均P〈0．01）。细胞增殖实验显示，对照组、1ng／mlrIL—1α组、10ng／mlrIL—1α组和100ng／mlrIL—1α组的吸光度值分别为1．37±0．18、1．79±0．14、2．14±0．17和2．21±0．23，IL—1α能够显著提高HUVEC的增殖能力，并与IL-1α浓度有关（均P〈0．叭）。IL—1α能够抑制HUVEC的迁移和增殖（均P〈0．01）。在共同培养系统中，对照组、HUVECs＋10pg／mlrIL—1α组和HUVECs＋HT-29组中血管内皮生长因子（VEGF）水平分别为（1．697±0．072）ng／ml、（3．507±0．064）ng／ml和（4．139±0.039）ng／ml，HUVECs＋10pg／mlrIL-1α组和HUVECs＋HT-29组分别与HUVECs组比较，差异均有统计学意义（均P〈0．01）。结论结肠癌细胞起源的IL-1α在结肠癌的转移过程中有重要作用，其通过结合IL-1RI提高结肠癌细胞VEGF的分泌水平，强化血管内皮细胞的增殖和迁移，进而促进肿瘤的新生血管；而IL-1α具有抑制IL-1α水平的作用，从而抑制结肠癌的新生血管。Objective To explore the effect of colon cancer cell-derived intedeukin-1α on the migration and proliferation of human umbilical vein endothelial cells as well as the role of IL-1α and IL-1α in the angiogenesis process. Methods Western blot was used to detect the expression of IL-1α and IL-1R1 protein in the colon cancer cell lines with different liver metastatic potential. We also examined how IL-1α and IL-1α influence the proliferation and migration of umbilical vascular endothelial cells assessed by PreMix WST-1 assay and migration assay, respectively. Double layer culture technique was used to detect the effect of IL-1α on the proliferation and migration of vascular endothelial cells and the effect of IL-1α on the vascular endothelial cells. Results Western blot analysis showed that IL-1α protein was only detected in highly metastatic colon cancer HT-29 and WiDr cells, but not in the lowly metastatic CaCo-2 and CoLo320 cells.Migration assay showed that there were significant differences in the number of penetrated ceils between the control （17.9±3.6） and 1 ng/ml rIL-1α group （23.2±4.2）, 10 ng/ml rIL-1α group （31.7±4.5）, and 100 ng,/ml rIL-1α group （38.6 ± 4.9）, showing that it was positively correlated with the increasing concentration of rIL-1α （P〈0.01 for all）. The proliferation assay showed that the absorbance values were 1.37±0.18 in the control group, and 1.79±0.14 in the 1 ng/ml rIL-1α group, 2.14±0.17 in the 10 ng/ml rIL-1α group, and 2.21 ±0.23 in the 100 ng/ml rIL-1α group, showing a positive correlation with the increasing concentration of rIL-1α （P〈0.01 for all）. IL-1α significantly inhibited the proliferation and migration of vascular endothelial cells （P〈0.01）. The levels of VEGF protein were （1.697±0.072） ng/ml, （3.507±0.064） ng/ml and （4.139±0.039） ng/ml in the control, HUVECs±IL-lot and HUVECs±HT-29 co- eultare system groups, respectively, showing a significant difference between the control and HUVECs ± 10 pg/ml rIL

关 键 词：血管内皮细胞增殖 肿瘤细胞起源 白细胞介素1Α 结肠癌细胞 细胞迁移 WESTERNBLOT法 rIL-1α HUVECS 

分 类 号：R735.35[医药卫生—肿瘤]