蛇床子素在体外对人肝癌细胞增殖和侵袭的抑制作用

The inhibiting effect of osthole on proliferation and invasion of hepatocellular carcinoma cells

目的:研究蛇床子素在体外对人肝癌HepG2、SMMC-7721、Bel-7402细胞增殖和侵袭的抑制作用。方法:取人肝癌细胞进行复苏,传代,培养。实验分为空白组和给药组,给药组给予不同浓度梯度(50、100、150、200μmol/L)蛇床子素药液处理。利用MTT法和细胞集落实验测定不同浓度蛇床子素体外抑制人肝癌细胞增殖作用;利用细胞划痕实验和Transwell小室实验测定不同浓度蛇床子素对人肝癌细胞的迁移和侵袭的抑制作用;利用蛋白质印迹(Western blot)法评价蛇床子素对人肝癌细胞中上皮细胞-间充质转化(EMT)相关蛋白E-cadherin和细胞迁移相关蛋白基质金属蛋白酶2(MMP-2)表达情况的影响。结果:蛇床子素能在体外抑制人肝癌细胞HepG2、SMMC-7721和Bel-7402的增殖,IC50值分别为147.62、141.57、179.67μmol/L,呈现一定的浓度依赖性。与空白组相比,给药组在蛇床子素50、75、100μmol/L浓度条件下可以显著抑制人肝癌细胞增殖(P<0.05)。与空白组相比,给药组人肝癌细胞的迁移和侵袭能力显著降低(P<0.05);给药组细胞MMP-2蛋白的表达量降低(P<0.05),E-cadherin蛋白的表达量增加(P<0.05)。结论:蛇床子素在体外对人肝癌细胞增殖和侵袭具有抑制作用,其抑制侵袭作用机制与下调MMP-2蛋白和激活E-cadherin蛋白的表达有关。

Objective:To investigate the effect of osthole on the proliferation and invasion of hepatocellular carcinoma cells HepG2,SMMC-7721,Bel-7402.Methods:Human liver cancer cells were taken for resuscitation,passage and culture.The experimental setting was divided into a blank group and an administration group.The administration group was treated with different concentrations of osthol liquid(50,100,150,200μmol/L).MTT method and cell colony experiment were used to determine the inhibitory effect of different concentrations of osthole on the proliferation of human liver cancer cells in vitro.Wound healing assay and transwell assay were used to investigate the inhibition of migration and invasion on HCC.Western blot was used to detect the influence of E-cadherin expression,a protein related to transformation(EMT)and MMP-2,a protein related to cell migration were detected by Western blot.Results:Osthole could inhibit the proliferation of HepG2,SMMC-77721 and Bel-7402 in vitro,with IC50 values of 147.62,141.57 and 179.67μmol/L,respectively,showing a certain concentration-dependent.Compared with the blank group,the treatment group could significantly inhibit the proliferation of human hepatocellular carcinoma cells under the conditions of 50,75,and 100μmol/L(P<0.05).It was found that osthole could effectively inhibit the migration and invasion of HCC.Compared with the blank group,the administration group showed statistically significant difference(P<0.05)The protein expression of MMP in the cells of the administration group decreased(P<0.05),while the expression of E-cadherin protein increased(P<0.05).Conclusion:Osthole can inhibit the proliferation and invasion in HCC and its mechanism may be related to protein E-cadherin and MMP-2.