维生素K缺乏或拮抗剂-Ⅱ诱导的蛋白质对肝癌细胞增殖和转移的影响及其机制

Effect of vitamin K deficiency or protein-induced protein Ⅱ on the proliferation and metastasis of hepatoma cells and its mechanism

目的 观察维生素K缺乏或拮抗剂-Ⅱ诱导的蛋白质（PIVKA-Ⅱ）对肝癌细胞增殖和转移的影响,并探讨其机制.方法 将SMMC-7721细胞加入PIVKA-Ⅱ培养,细胞计数试剂盒（CCK-8）法检测细胞增殖,平板克隆形成实验测定SMMC-7721细胞克隆形成,流式细胞仪检测细胞凋亡,Transwell法检测细胞迁移能力,Western blot实验测定细胞转化生长因子-α（TGF-α）、碱性纤维母细胞生长因子（bFGF）、血管内皮生长因子（VEGF）、基质金属蛋白（MMP）-2和MMP-9表达.结果 PIVKA-Ⅱ组SMMC-7721细胞克隆形成率（1.94±0.14）和细胞存活率（1.53±0.21）均高于阴性对照组（1.12±0.08、1.11±0.15）和空白对照组（1.00±0.03、1.00±0.04;F=68.254、10.264,P=0.000、0.000）.PIVKA-Ⅱ组SMMC-7721细胞凋亡率（1.52±0.28）低于阴性对照组（3.13±0.36）和空白对照组（3.45±0.43;F=48.436,P=0.000）.PIVKA-Ⅱ组SMMC-7721细胞迁移数[（62.54±7.45）个]和迁移率（2.13±0.21）均高于阴性对照组[（31.34±6.03）个、1.06±0.03]和空白对照组[（29.42±5.59）个、1.00±0.05;F=46.241、283.240,P=0.000、0.000].PIVKA-Ⅱ组SMMC-7721细胞TGF-α、bFGF、VEGF、MMP-2和MMP-9的表达量（1.69±0.13、1.30±0.04、1.63±0.11、1.72±0.23、1.88±0.54）均高于阴性对照组（1.11±0.07、1.06±0.01、1.08±0.02、1.14±0.06、1.17±0.04）和空白对照组（1.00±0.01、1.00±0.01、1.00±0.02、1.00±0.01、1.00±0.01;F=2014.254、476.355、1845.364、6.486、7.045,P=0.000、0.000、0.000、0.000、0.000）.结论 PIVKA-Ⅱ 能够促进肝癌细胞增殖和迁移,抑制肝癌细胞凋亡,其机制可能和PIVKA-Ⅱ能够促进TGF-α、bFGF、VEGF、MMP-2和MMP-9表达有关.

Objective To investigate the effect of vitamin K deficiency or protein-induced protein Ⅱ （PIVKA-Ⅱ） on the proliferation and metastasis of hepatoma cells and to explore its possible mechanism.Methods The SMMC-7721 cells were cultured with PIVKA-Ⅱ.The cell proliferation was assayed by cell counting kit-8 （CCK-8）.The SMMC-7721 cell clone formation was assayed plate clone formation.The cell apoptosis was assayed by flow cytometry.The cell migration was assayed by transwell.The expression of transforming growth factor-α （TGF-α）, basic fibroblast growth factor （bFGF）, vascular endothelial growth factor （VEGF）, matrix metalloproteinase （MMP）-2 and MMP-9 were detected by Western blotting.Results The cell clone formation rate （1.94±0.14） and cell survival rate （1.53±0.21） in PIVKA-Ⅱ group was significantly higher than those in negative control group （1.12±0.08, 1.11±0.15） and blank control group （1.00±0.03, 1.00±0.04;F=68.254, 10.264, P=0.000, 0.000）.The apoptosis rate of PIVKA-Ⅱ group（1.52±0.28） was lower than that of negative control group （3.13±0.36） and blank control group （3.45±0.43;F=48.436, P=0.000）.The cell migration number （62.54±7.45） and mobility （2.13±0.21） of PIVKA-Ⅱ group was significantly higher than those in negative control group （31.34±6.03, 1.06±0.03） and blank control group （29.42±5.59, 1.00±0.05;F=46.241, 283.240, P=0.000, 0.000）.The expression of TGF-α, bFGF,VEGF, MMP-2,MMP-9 （1.69±0.13, 1.30±0.04, 1.63±0.11） in PIVKA-Ⅱ group was higher than that in negative control group （1.11±0.07, 1.06±0.01, 1.08±0.02,1.14±0.06, 1.17±0.04） and blank control group （1.00±0.01, 1.00±0.01, 1.00±0.02,1.00±0.01, 1.00±0.01;F=2 014.254, 476.355, 1 845.364, 6.486, 7.045, P=0.000, 0.000, 0.000, 0.000, 0.000）.Conclusion PIVKA-Ⅱ can promote the proliferation and migration of hepatocellular carcinoma cells and inhibit the apoptosis of hepatoma cells.The mechanism may be related to that PIVKA-Ⅱ can promote the expression of TGF-α, bFGF, VEGF, MMP-2 and MMP-9.