槲皮素对人腺样囊性癌ACC-M细胞增殖和凋亡的影响及机制研究

Effects and mechanism of quercetin on proliferation and apoptosis of human adenoid cystic carcinoma cell line ACC-M

目的:观察槲皮素对人腺样囊性癌细胞系ACC-M增殖和凋亡的作用,探讨槲皮素影响ACC-M细胞凋亡的可能机制。方法:采用CCK-8法检测不同浓度的槲皮素对ACC-M细胞增殖的抑制作用并计算抑制率,计算IC_(50);流式细胞术检测ACC-M细胞凋亡率和细胞周期分布;实时荧光定量RT-PCR检测ACC-M细胞中Survivin mRNA的表达。采用SPSS 20.0软件包对数据进行统计学分析。结果:槲皮素抑制ACC-M细胞增殖的作用明显,且呈浓度依赖性,IC_(50)为151.12μmol/L(P<0.05);Annexin V/PI双染法检测显示,槲皮素能够诱导ACC-M细胞凋亡,且与剂量、时间呈正相关(P<0.05);PI染色法检测显示,槲皮素主要作用于G2/M节点,能将细胞特异性地阻滞于G2/M期;RTPCR检测显示,随着药物浓度增加,ACC-M细胞中Survivin mRNA的表达水平均呈不同程度的降低,200μmol/L槲皮素可显著抑制Survivin mRNA的表达。结论:槲皮素能够抑制ACC-M细胞增殖,诱导其凋亡,且呈时间-剂量依赖性。槲皮素可能通过下调Survivin的表达水平而发挥抗腺样囊性癌的作用。

PURPOSE ： To observe the effect of quercetin on human adenoid cystic carcinoma cells （ACC-M） proliferation and apoptosis. METHODS ： CCK-8 staining was used to detect different concentrations of quercetin on ACC-M cell proliferation inhibition, the inhibition rate and IC50 were calculated; flow cytometry was used to detect ACC-M cell apoptosis rate and cell cycle distribution; Survivin mRNA of ACC-M cells was detected by RT-PCR. The difference between each group was analyzed with SPSS20.0 software package. RESULTS ： Quercetin inhibited ACC-M cell growth obviously in a concentration dependent manner. The IC50 was 151.12 μmol/L （P〈0.05）; annexin V/PI double staining assay showed that quercetin can induce apoptosis of ACC-M cells in a dose and time dependent manner （P〈0.05）; PI staining assay showed that quercetin played an important role in G2/M cells, which were specifically arrested in G2/M phase. RT-PCR showed the quercetin concentration was negatively related with survivin mRNA in ACC-M cells and 200 μmol/L quercetin could inhibit the expression of Survivin mRNA significantly. CONCLUSION S： Quercetin can inhibit the proliferation of ACC-M cells and induce its apoptosis in a time and dose dependent manner by down-regulating the level of Survivin.