壁虎超滤组分对人肝癌HepG2细胞增殖及迁移的影响

Effects of gecko ultrafiltration components on proliferation and migration of human hepatoma HepG2 cells

目的探讨壁虎超滤组分(GUCs)对人肝癌HepG2细胞的增殖及迁移的影响。方法通过噻唑蓝法检测细胞的增殖,选定分组浓度,将人肝癌HepG2细胞分为空白组(不加药)、对照组(10μg·mL^-15-氟尿嘧啶)和低、中、高浓度实验组(0.10,0.15和0.225 mg·mL^-1 GUCs)。培养24 h后,用Transwell实验检测细胞的迁移能力,用蛋白质免疫印迹法检测血管内皮生长因子(VEGF)、Rac家族小GTP酶1(Rac-1)和E钙黏蛋白(E-cadherin)蛋白表达水平的变化。结果GUCs能显著抑制肝癌HepG2细胞增殖,呈时间-剂量依赖性。24,48和72 h的半数抑制浓度分别为0.24,0.21和0.18 mg·mL^-1。空白组、对照组和低、高浓度实验组的透膜细胞数分别为(131.25±16.79),(68.50±2.41),(69.25±14.16)和(12.25±2.35)个,E-cadherin蛋白灰度值分别为0.13±0.01,0.23±0.01,0.20±0.02和0.27±0.02,VEGF蛋白灰度值分别为0.84±0.07,0.74±0.09,0.73±0.06和0.31±0.01,Rac-1蛋白灰度值分别为0.40±0.05,0.33±0.02,0.32±0.02和0.27±0.02,低、高浓度实验组和对照组的上述指标与空白组比较,差异均有统计学意义(均P<0.05)。结论GUCs能抑制HepG2细胞增殖以及迁移,可能与上调E-cadherin基因表达、下调VEGF和Rac-1基因表达有关。

Objective To explore the effects of gecko ultrafiltration components(GUCs)on the proliferation and migration of human liver cancer HepG2 cells.Methods The cell proliferation was detected by the methyl thiazolyl tetrazolium method,and the group concentration was selected to divide the human liver cancer HepG2 cells into blank group(no drug),control group(10μg·mL^-15-fluorouracil)and experimental-L,-M,-H groups(0.10,0.15 and 0.225 mg·mL^-1 GUCs).After 24 h incubation,Transwell test was used to detect the migration ability of cells,and Western Blotting was used to detect the changes of protein expression level of vascular endothelial growth factor(VEGF)and Rac family small GTPase 1(Rac-1)and E-cadherin.Results GUCs can significantly inhibit the proliferation of liver cancer HepG2 cells in a dose-dependent manner.The median inhibitory concentrations at 24,48 and 72 h were 0.24,0.21 and 0.18 mg·mL^-1,respectively.The number of permeable cells in the blank group,control group and experimental-L,-H groups were(131.25±16.79),(68.50±2.41),(69.25±14.16)and(12.25±2.35),the gray value of E-cadherin protein were 0.13±0.01,0.23±0.01,0.20±0.02 and0.27±0.02,the gray value of VEGF protein were 0.84±0.07,0.74±0.09,0.73±0.06 and 0.31±0.01,the gray value of Rac-1 protein were 0.40±0.05,0.33±0.02,0.32±0.02 and 0.27±0.02,compared with the blank group,the above indexes of experimental-L,-H groups and control groups had statistically significant differences(all P<0.05).Conclusion GUCs can inhibit the proliferation and migration of HepG2 cells,which may be related to the up-regulation of E-cadherin gene expression and down-regulation of VEGF and Rac-1 gene expression.