中电导钙激活K+通道对肝癌细胞迁移的影响

Effects of the intermediate conductance calcium-actived K+channel on migration of hepatocellular carcinoma cells

目的研究阻断中电导钙激活K+通道(KCa3.1)对人源肝癌细胞(HepG2)迁移的影响。方法将人源肝癌细胞HepG2分为3组:空白组、TRAM-34(钙激活钾通道特异性阻断药)组和联合组。空白组细胞不做处理,TRAM-34组用30μmol·L^-1TRAM-34处理48 h,联合组用30μmol·L-1 TRAM-34+100μmol·L^-1EBIO(钙激活钾通道特异性激动药)共同处理48 h。用蛋白质印迹法检测Kca3.1蛋白在HepG2细胞中的表达;用细胞迁移侵袭实验和细胞划痕实验检测HepG2细胞的迁移能力。结果KCa3.1钾通道蛋白在HepG2与LO2细胞(正常肝细胞)中的相对表达量为1.49±0.24,0.74±0.16,两者间比较,差异有统计学意义(P<0.05)。空白组、TRAM-34组和联合组的HepG2细胞的迁移能力(Transwell实验,光密度值)分别为0.28±0.01,0.20±0.04和0.26±0.05;TRAM-34组与空白组、联合组相比,差异均有统计学意义(均P<0.05)。TRAM-34可明显抑制HepG2细胞的迁移能力,但EBIO可部分抵消此抑制作用;划痕实验也证实了这一趋势。结论阻断KCa3.1钾通道可抑制HepG2细胞的迁移。

Objective To investigate the effect of blocking the K+channel(KCa3.1)activated by calcium on the migration of human hepatocellular carcinoma(HepG2)cells.Methods Human hepatocellular carcinoma cells HepG2 were divided into 3 groups:Blank group,TRAM-34(triarylmethane-34)group and joint group[30μmol·L^-1 TRAM-34+100μmol·L^-1 EBIO(2H-benzimidazol-2-one)],for 48 h.Western blot was used to detect the expression of Kca3.1 protein in HepG2 cells.Migration ability of HepG2 cells was detected by cell migration and invasion test(optical density,OD value)and cell scratch test.Results The relative expression of KCa3.1 potassium channel in HepG2 and LO2 cells were 1.49±0.24,0.74±0.16;there was statistically significant difference between two cells(P<0.05).The ability to migrate after Transwell in blank group,TRAM-34 group and joint group were 0.28±0.01,0.20±0.04 and 0.26±0.05,respectively.Compared with blank group and joint group,the results in the TRAM-34 group were statistically significant(P<0.05).TRAM-34 significantly inhibited the migration of HepG2 cells,but EBIO partially offset the inhibition.The scratch test also well confirmed this trend.Conclusion Blocking KCa3.1 potassium channel can inhibit the migration of HepG2 cells.