IL-8通过整合素调控肝癌HepG2细胞迁移

Integrins mediate the migration of HepG2 cells induced by IL-8

目的研究IL-8对肝癌细胞HepG2增殖、迁移的影响,探讨整合素α亚基在调控HepG2细胞迁移中的作用。方法以不同浓度（0-125 ng/ml）的IL-8刺激HepG2细胞,分别采用MTT法检测IL-8对肝癌细胞的增殖作用;细胞划痕损伤模型测定不同处理组在0、4、8、12和24 h各时间点对HepG2细胞水平迁移的影响;Transwell法检测刺激24 h后HepG2细胞纵向迁移能力;免疫荧光检测0、125 ng/ml IL-8刺激HepG2细胞8 h后细胞骨架的变化;Western印迹测定整合素α亚基的表达变化规律。结果与对照组相比,IL-8促进HepG2细胞增殖,但各浓度间无明显差异;细胞划痕损伤实验和Transwell实验均表明IL-8可促进HepG2细胞迁移,且具有浓度依赖性;IL-8诱导HepG2细胞骨架重排,使丝状伪足样凸起数目增多;Western印迹结果显示,IL-8上调整合素α亚基的表达,但各亚基具有浓度性差异。结论 IL-8可通过上调整合素α亚基的表达促进HepG2细胞迁移,各α亚基调控作用可能具有差异性。

Objective To investigate the effect of IL-8 on the viability and migration of HepG2 cells and the potential effect of integrins（ α subunits） on the migration of HepG2 cells. Methods HepG2 cells were stimulated by IL-8 at different concentrations ranging from 0 to 125 ng / ml in vitro. MTT assay was preformed to detect the viability of HepG2 cells. Scratch wound migration assay was used to explore the effect of IL-8 on the migration of HepG2 cells at the time points of 0,4,8,12 and 24 h,respectively. Transwell assay was conducted to detect the vertical migration of HepG2 cells after stimulation with IL-8 for 24 h. The F-actin of HepG2 cells was observed by immunofluorescence analysis after treatment with IL-8 at different concentrations. The expression of α subunits of integrins in HepG2 cells treated with IL-8for 8h was detected using Western boltting assays. Results Compared with the control group,HepG2 cells treated with IL-8 showed improved proliferation activities,but there was no significant difference between the groups of HepG2 cells treated with IL-8 at different concentrations. The cell scratch healing assays and Transwell analysis both indicated that IL-8promoted the migration of HepG2 cells in a dose-dependent manner. Meanwhile,the cytoskeleton of HepG2 cells was rearranged and the number of filopodium-like protrusions（ FLPs） was increased rapidly after treatment with IL-8. Western blotting analysis showed that IL-8 up-regulated the expression of α subunit of integrins and there was a concentration difference between α subunits. Conclusion IL-8 promotes the migration of HepG2 cells by up-regulating the expression ofα subunits,and different α subunits may play different roles.