论文部分内容阅读
目的:研究蛇葡萄素钠(AMP-Na)对人肝癌HepG2细胞株的抗肿瘤作用,探讨其抗肿瘤作用机制。方法:应用MTT法测定AMP-Na对HepG2细胞的抑制作用,并计算IC50值;流式细胞术检测AMP-Na的诱导凋亡作用和对细胞周期的影响;实时荧光定量PCR检测AMP-Na对HepG2细胞CyclinD1基因mRNA的表达影响。结果:MTT检测结果表明,AMP-Na显著抑制HepG2细胞的增殖,呈现时间和剂量依赖效应,药物作用24h、48h、72h IC50值分别为226μg/ml、67μg/ml和34μg/ml。流式细胞仪检测表明100μg/ml AMP-Na作用于细胞48 h、72 h凋亡率分别为31.9%、34.0%;可使G1期细胞比例增加,S期细胞比例减少,有明显的G1期阻滞作用;实时荧光定量PCR检测AMP-Na 100μg/ml作用HepG2细胞48 h、72 h后,用药组CyclinD1 mRNA表达分别是对照组的0.551和0.524倍,二者均有显著性差异(P<0.05)。结论:AMP-Na对肝癌HepG2细胞有抗肿瘤效果,其机理可能通过降低细胞周期调节关键的CyclinD1的表达,使细胞停滞于G1期,诱导细胞凋亡而发挥抗肿瘤作用。
Objective: To study the anti-tumor effect of AMP-Na on human hepatocellular carcinoma HepG2 cell line and its anti-tumor mechanism. Methods: The inhibitory effect of AMP-Na on HepG2 cells was determined by MTT method and the IC50 values were calculated. The apoptosis induced by AMP-Na and the cell cycle were detected by flow cytometry. The expression of AMP-Na was detected by real-time fluorescence quantitative PCR Effect of CyclinD1 gene mRNA expression in HepG2 cells. Results: The results of MTT assay showed that AMP-Na significantly inhibited the proliferation of HepG2 cells in a time and dose-dependent manner. The IC50 values of the drugs for 24h, 48h and 72h were 226μg / ml, 67μg / ml and 34μg / ml respectively. The results of flow cytometry showed that the apoptotic rates of AMP-Na at 100 μg / ml for 48 h were 31.9% and 72.3%, respectively. The proportion of cells in G1 phase increased and the proportion of S phase cells decreased. The expression of CyclinD1 mRNA in HepG2 cells treated with 100μg / ml AMP-Na for 48 h and 72 h were 0.551 and 0.524 times of the control group, respectively, both of which were significantly different (P < 0.05). CONCLUSION: AMP-Na has anti-tumor effect on HepG2 hepatocarcinoma cells. The possible mechanism is that AMP-Na may play an antitumor role by decreasing the expression of CyclinD1, which is the key cell cycle regulator, arresting cells in G1 phase and inducing apoptosis.