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

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IC₅₀;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,IC₅₀为151.12 μmol/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;RT-PCR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,200 μmol/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。

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

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.