白细胞介素-33对肝癌细胞迁移、侵袭能力和细胞骨架的影响

Effects of interleukin-33 on the invasion,migration and cytoskeleton of hepatoma carcinoma cell

目的:探讨白细胞介素(IL)-33对肝癌细胞迁移能力、侵袭能力和细胞骨架的影响及其作用机制。方法:通过蛋白质印迹法检测肝癌细胞中β-链蛋白、基质金属蛋白酶(MMP)-2以及MMP-9蛋白表达水平,流式细胞仪分析肝癌细胞表面抗原分子的表达水平,附壁试验和细胞聚集试验分别检测肝癌细胞对细胞外基质的亲和力以及肝癌细胞的聚集能力,细胞迁移划痕实验和穿透小室法测定IL-33对肝癌细胞体外迁移侵袭能力的影响,高内涵细胞分析系统免疫荧光法检测IL-33对肝癌细胞骨架的影响。结果:在IL-33的作用下,肝癌细胞中β-链蛋白、MMP-2以及MMP-9蛋白的表达均显著下降(P<0.05),并有剂量和时间依赖性。与对照组相比,经IL-33作用后肝癌细胞间黏附分子-1、人类白细胞抗原-Ⅰ的表达率显著增加,且CD44表达显著下降(P<0.05);肝癌细胞对细胞外基质的亲和力及肝癌细胞聚集程度均降低(P<0.05);实验组肝癌细胞的微丝数量和细胞骨架的面积显著降低,且与剂量相关(P<0.05)。结论:IL-33能够有效抑制肝癌细胞的迁移和侵袭,并且有降低细胞骨架微丝数量和面积的作用。

Objective： To investigate the mechanism and effects of interleukin-33（IL-33） on the migration,invasion and cytoskeleton of human hepatoma carcinoma cells. Methods： The protein expression levels of β-catenin,matrix metalloproteinase-2（MMP-2） and matrix metalloproteinase-9（MMP-9） in hepatoma carcinoma cells were detected by Western blotting. The expression of antigenic molecules was analyzed by flow cytometer. The tumor cell binding affinity to extracellular matrix components and the degree of homotypic aggregation were measured by cell attachment and aggregation assay,respectively. The invasive and migratory abilities of IL-33 were detected by Transwell migration assay and wound healing assay,respectively. The effect of IL-33 on cytoskeleton was evaluated by high intension cell analysis system. Results： The expression of β-catenin,MMP-2 and MMP-9 were significant decreased in experimental group than those in control group in a dose and time dependent manner after the IL-33 treatment（P 〈 0. 05）. Compared with control group,the expression of ICAM-1,HLA-Ⅰ increased significantly while the CD44 was significantly decreased（P 〈 0. 05）. And the binding affinity to extracellular matrix components and the degree of homotypic aggregation were significantly decreased after IL-33 treatment. The microfilament cytoskeleton and the area of cytoskeleton decreased more significantly than those in control group（P 〈 0. 05）. Conclusion：The IL-33 can effectively suppress the invasion and migration of hepatoma carcinoma cell,and reduce the amount of microfilament and the area of cytoskeleton.