P-5m八肽对肝癌HepG2细胞中MMP-2和MMP-9表达的抑制作用

摘要

Objective To explore the impact of P-5m octapeptide on the expressions of matrix metalloproteinase-2( MMP-2) and matrix metalloproteinase-9( MMP-9) in HepG2 cells,and to investigate the antitumor mechanism of P-5m octapeptide further.Methods The HepG2 cells in logarithmic growth phase were stimulated by P-5m octapeptide for 36 h at the concentrations of 10 μmol/L and 100 μmol/L, respectively.Then, the cells were treated after 36 h.The expressive variations of MMP-2 and MMP-9 mRNA were detected by Real-time PCR,and the changes of the expressions of MMP-2 and MMP-9 proteins were detected by western blot assay.Results Real-time PCR analysis showed that there were significant differences on the expressions of MMP-2 mRNA and MMP-9 mRNA compared with the control group(P<0.05),after the cells were stimulated by P-5m octapeptide at the concentrations of 10 μmol/L and 100 μmol/L for 36 h, respectively.Western blot analysis showed that P-5m octapeptide could inhibit the expressions of MMP-2 protein and MMP-9 protein in HepG2 cells compared with the control group(P<0.05)after 36 h stimulation at the concentrations of 10 μmol/L and 100μmol/L,respectively.Conclusion P-5m octapeptide can remarkably inhibit the expressions of MMP-2 and MMP-9 in HepG2 cells.%目的 探讨P-5m八肽对肝癌HepG2细胞中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达的影响,以及P-5m八肽的抗肿瘤作用机制.方法 用终浓度为10μmol/L和100μmol/L的P-5m八肽分别对处于对数生长期的HepG2细胞进行处理,药物作用时间为36 h,36 h后将细胞裂解.采用Real-time PCR方法检测给药后细胞中MMP-2和MMP-9 mRNA表达水平的变化;Western blot方法测定MMP-2和MMP-9蛋白表达水平的变化.结果 Real-time PCR检测显示终浓度为10μmol/L和100μmol/L的P-5m八肽对HepG2细胞刺激后,MMP-2和MMP-9的mRNA表达均受到抑制,与对照组比较差异具有统计学意义(P<0.05);Western blot检测结果表明:用10μmol/L和100μmol/L的P-5m八肽对HepG2细胞刺激后,MMP-2和MMP-9的蛋白表达均受到抑制,与对照组比较差异具有统计学意义(P<0.05).结论 P-5m八肽可明显抑制HepG2细胞中MMP-2和MMP-9的表达.