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目的探讨在外源性层黏连蛋白(LN)与其受体结合下,姜黄素对人肝癌HepG2细胞生长、凋亡的影响。方法实验分为对照组、LN组(20μg/L LN)、姜黄素组(40μmol/L姜黄素)、联合组(20μg/L LN+40μmol/L姜黄素)。采用酸性磷酸酶法(APA)、流式细胞术(FCM)和Western blotting法等探讨在外源性LN与其受体结合下,姜黄素对人肝癌HepG2细胞生长、细胞凋亡率、线粒体膜电位、细胞增殖相关蛋白α-蛋白激酶(α-PKC)及细胞凋亡相关蛋白人多聚ADP核糖聚合酶(PARP)、Caspase-3、Bcl-2和p53表达的影响。结果 LN组与对照组相比,细胞存活率增加。姜黄素组与联合组能显著抑制人肝癌HepG2细胞的增殖,并呈时间依赖性。姜黄素组与联合组作用48h时,倒置显微镜下,细胞数量明显减少,皱缩变圆,大部分细胞悬浮;中晚期凋亡和坏死细胞比率(%)分别为97.04±1.50,98.02±1.35;细胞内钙离子浓度升高;线粒体膜电位下降;增殖相关蛋白α-PKC含量减少;凋亡相关蛋白PARP出现剪切带,Caspase-3表达下调,p53表达上调,Bcl-2表达无明显变化。结论在外源性层黏连蛋白与其受体结合下,姜黄素与人肝癌HepG2细胞作用后抑制生长并诱导细胞发生凋亡;显示出姜黄素抗肿瘤作用稳定,其机制可能与上调p53,下调α-PKC有关。
Objective To investigate the effect of curcumin on the growth and apoptosis of human hepatocellular carcinoma HepG2 cells induced by exogenous laminin (LN) combined with its receptor. Methods The experiment was divided into control group, LN group (20μg / L LN), curcumin group (40μmol / L curcumin) and combination group (20μg / L LN + 40μmol / L curcumin). The effects of curcumin on HepG2 cell growth, apoptosis rate, mitochondrial membrane potential, apoptosis rate and apoptosis were investigated by acid phosphatase (APA), flow cytometry (FCM) and Western blotting. Effects of α - PKC and PARP, Caspase - 3, Bcl - 2 and p53 on the Expression of Apoptosis Related Protein. Results LN group compared with the control group, the cell survival rate increased. Curcumin group and combination group can significantly inhibit the proliferation of HepG2 cells in a time-dependent manner. Curcumin group and the combined group 48h, under inverted microscope, the number of cells was significantly reduced, shrinkage round, most of the cells suspended; late apoptosis and necrosis cells (%) were 97.04 ± 1.50,98.02 ± 1.35; The concentration of intracellular calcium increased; the mitochondrial membrane potential decreased; the content of a-PKC decreased; the apoptosis-related protein PARP appeared cut-off band, the expression of Caspase-3 was down-regulated and p53 was up-regulated. Conclusions Curcumin inhibits the growth of human hepatocellular carcinoma HepG2 cells and induces cell apoptosis under the binding of exogenous laminin and its receptor. Curcumin shows a stable antitumor effect, which may be related to the up-regulation of p53 and down-regulation of α -PKC related.