细胞外泛素对肝癌细胞增殖、侵袭和迁移能力的影响

Effect of extracellular ubiquitin on the proliferation,invasion,and migration of hepatoma cells

目的通过体外细胞实验,探讨细胞外泛素对肝细胞癌细胞增殖、侵袭和迁移能力的影响。方法利用不同浓度的细胞外泛素(200、400、800 ng/ml),分别干预肝癌细胞(HepG2),采用CCK8法检测细胞的增殖情况;利用Transwell法检测不同浓度泛素对肝癌细胞侵袭能力的影响;利用划痕实验和蛋白免疫印迹检测细胞外泛素对肝癌细胞迁移能力的作用。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果CCK8法检测结果显示,细胞外泛素干预后能明显促进肝癌细胞的增殖能力,并具有明显的浓度依赖性,且泛素干预浓度在400 ng/ml时作用最显著,在48、72、96 h相对吸光度值均明显高于对照组(P值均<0.05)。Transwell实验结果显示,与对照组相比,不同浓度的细胞外泛素均可明显促进肝癌细胞的侵袭,且在400 ng/ml泛素干预组最显著[(134.00±8.18)个vs(347.33±18.90)个,t=17.94,P<0.001]。划痕实验和蛋白免疫印迹结果显示,400 ng/ml泛素能显著增加HepG2肝癌细胞的迁移能力。结论细胞外泛素在体外能明显促进HepG2肝癌细胞的增殖、侵袭和迁移,且具有浓度依赖性。

Objective To investigate the effect of extracellular ubiquitin on the proliferation,invasion,and migration of hepatoma cells by in vitro cell experiments.Methods The hepatoma cells(HepG2)were treated with extracellular ubiquitin at different concentrations(200,400,and 800 ng/ml);CCK-8 assay was used to observe cell proliferation,Transwell assay was used to observe the effect of extracellular ubiquitin at different concentrations on the invasion ability of hepatoma cells,and wound healing assay and Western blotting were used to observe the effect of extracellular ubiquitin on the migration ability of hepatoma cells.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results The CCK-8 assay showed that extracellular ubiquitin significantly promoted the proliferation of hepatoma cells in a concentration-dependent manner,with the most significant effect at the concentration of 400 ng/ml,and the intervention group had a significantly higher relative absorbance value than the control group at 48,72,and 96 hours(all P<0.05).The Transwell assay showed that different concentrations of extracellular ubiquitin significantly promoted the invasion of hepatoma cells,with the most significant effect in the group treated by 400 ng/ml extracellular ubiquitin(134.00±8.18 vs 347.33±18.90,t=17.94,P<0.001).The Wound healing assay and Western blotting showed that 400 ng/ml extracellular ubiquitin significantly increased the migration ability of HepG2 cells.Conclusion Extracellular ubiquitin can significantly promote the proliferation,invasion,and migration of HepG2 cells in vitro in a concentration-dependent manner.