蛇葡萄素通过Dermcidin蛋白调节肝癌细胞株HepG2侵袭活力及侵袭基因表达的实验

Ampelopsis Regulates Invasive Activity and Invasion Gene Expression of Hepatocellular Carcinoma HepG2 Cell Line Through Dermcidin Protein

目的研究蛇葡萄素(AMP)对肝癌细胞株HepG2侵袭活力及侵袭基因表达的调节作用及分子机制。方法培养肝癌细胞株HepG2后用不同剂量的AMP处理(20、40、60、80μmol/L)、转染Dermcidin的siRNA,测定细胞的迁移能力、侵袭能力以及Dermcidin、侵袭基因mRNA表达量。结果不同剂量AMP处理后细胞的迁移、侵袭数目及细胞中Dermcidin、基质金属蛋白酶(MMP)2、MMP9、MMP10mRNA水平均低于对照组,且AMP剂量越大,细胞的迁移、侵袭数目及细胞中Dermcidin、MMP2、MMP9、MMP10mRNA水平越低;80μmol/L AMP联合dermcidinsiRNA后,HepG2细胞的迁移、侵袭数目以及细胞内MMP2、MMP9、MMP10mRNA表达水平均增多。结论AMP通过下调Dermcidin蛋白来抑制肝癌细胞株HepG2的侵袭活力、降低侵袭基因的表达水平。

Objective To investigate the regulatory effect of ampelopsin(AMP)on invasion activity and invasion gene expression of hepatocellular carcinoma HepG2 cell line and its molecular mechanism.Methods Hepatocellular carcinoma HepG2 cell line were cultured and treated with different doses of AMP(20,40,60,80μmol/L),and transfected with Dermcidin siRNA.Then the cell migration ability and invasive ability,the mRNA expression of Dermcidin and invasive genes were determined.Results After different doses of AMP treatment,the number of migration and invadsion cells and the mRNA levels of Dermcidin,matrix metalloproteinase(MMP)2,MMP9 and MMP10 were lower than those in the control group;the greater the AMP dose was,the lower the number of cell migration,the number of invasion and the mRNA levels of Dermcidin,MMP2,MMP9,MMP10 were;the number of migration and invasion cells and the mRNA levels of MMP2,MMP9,MMP10 in HepG2 cells treated with 80μmol/L AMP combined with dermcidin gene siRNA transfection were significantly increased.Conclusion AMP inhibits the invasive activity and decreases the invasive genes expression in HepG2 cell line by down-regulating the expression of Dermcidin protein.