pEGFP-N1介导BTI的表达抑制肝癌HepG2细胞迁移及细胞周期进程

pEGFP-N1-Mediated BTI Expression Inhibits Migration and Cell Cycle Progression in Hepatocellular Carcinoma HepG2 Cells

荞麦是一种药食同源的粮食作物,研究发现荞麦胰蛋白酶抑制剂(BTI)可以抑制肿瘤细胞的增殖。该文采用激光共聚焦显微镜、MTT法、划痕法、流式细胞术、qRT-PCR、Western blot等方法检测了pEGFP-N1介导的BTI在HepG2细胞内的表达对细胞的生长、黏附、迁移、凋亡及细胞周期的作用。结果显示,pEGFP-N1介导的BTI在HepG2细胞内的表达显著抑制了细胞生长、黏附和迁移,并使HepG2细胞凋亡率增加。同时E-钙黏蛋白的表达水平增高,MMP-2和MMP-9的表达下降。伴随着p53、p21、p63、p73的上调表达,周期蛋白CyclinD1、CyclinE1和周期依赖性激酶(CDK2、CDK4、CDK7)的表达下调,细胞周期阻滞在G1期。

Buckwheat,a specialty grain in China,is a common component of food products and medicines.BTI (Buckwheat trypsin inhibitor) reportedly inhibits tumor cells growth.Laser confocal microscopy,MTT assay,wound healing assay,flow cytometry,qRT-PCR,and Western blot were used to investigate the effect of cell viability,adhesion,migration,apoptosis,and cell cycle after pEGFP-N1-BTI transfected in human hepatocellular carcinoma HepG2 cells.The pEGFP-N1-mediated BTI expression significantly inhibited proliferation,adhesion,and migration of HepG2 cells.Meanwhile,the cell apoptosis rates were increased and the cell cycle was arrested.Moreover,pEGFP-N1-mediated BTI enhanced the protein expression level of E-cadherin and decreased those of MMP-2 and MMP-9.The potential mechanism involved the following:p53,p21,p63,and p73 up-regulation;CyclinD1,CyclinE1,and cyclin-dependent kinases (CDK2,CDK4,and CDK7) down-regulation;and arrest of the HepG2 cells cycle in G1 phase.