达玛-24-烯-3β-乙酰氧基-20s-醇对胃癌细胞增殖的抑制作用

发布时间：2018-03-30 00:31

本文选题：达玛--烯-β-乙酰氧基-s-醇　切入点：胃癌细胞　出处：《医学研究生学报》2017年05期

【摘要】：目的前期研究发现中药连翘醇提物具有明显体外抑制肿瘤细胞的作用,文章进一步探讨从中药连翘提取的化合物达玛-24-烯-3β-乙酰氧基-20s-醇(Dammar-24-ene-3β-acetate-20S-ol,DM)体外抗肿瘤作用及其机制。方法采用MTT法检测DM对胃癌细胞株SGC-7901、BGC-823和MKN-45体外抑制作用。实验分为MKN-45对照组及其低、高剂量组,BGC-823对照组及其低、高剂量组,SGC-7901对照组及其低、高剂量组,对照组均为新鲜培养基,低剂量DM为10、50μg/m L,流式细胞仪检测凋亡细胞比例。应用Cell Quest软件分析结果,记录细胞不同周期的比例。采用DCFH-DA探针检测ROS水平,实验分为空白对照组,多西紫杉醇组和DM组。配置微管聚集实验反应体系,其中多西紫杉醇10μmol/L,DM终浓度选择50或100μmol/L,空白对照组不加药物,采用微管聚集实验和微管蛋白免疫荧光染色研究DM对于微管系统的影响。结果 50μg/m L对于3株胃癌细胞抑制率均在80%以上,IC50分别为:MKN-45(11.72±1.35)μg/m L;BGC-823(17.19±0.82)μg/m L;SGC-7901(7.55±0.79)μg/m L。2μg/m L DM处理48 h后再以低密度培养8 d,细胞克隆较对照组明显减少。与MKN-45对照组凋亡细胞[(21.1±2.5)%]相比,MKN-45低剂量组和高剂量组凋亡细胞比率[(25.1±1.3)%和(55.2±2.3)%]均有明显升高(P0.01)。BGC-823对照组凋亡细胞百分率(13.2±2.4)%比较,BGC-823低、高剂量组凋亡细胞百分率[(18.2±2.1)%、(41.8±2.8)%]明显升高(P0.01)。与SGC-7901对照组凋亡细胞率[(10.5±1.8)%]比较,SGC-7901高剂量DM组凋亡细胞率[(21.1±1.9)%]升高(P0.05),而与SGC-7901低剂量DM组[(12.3±1.6)%]差异无统计学意义(P0.05)。与MKN-45对照组比较,MKN-45低剂量组S期细胞百分率降低[(14.5±2.7)%vs(12.3±3.3)%,P0.05]。与BGC-823对照组比较,BGC-823低剂量组细胞S期百分率增加[(12.2±5.4)%vs(20.2±2.1)%,P0.05]。与SGC-7901对照组比较,SGC-7901低剂量组S期细胞百分率增加[(21.5±3.8)%vs(31.3±2.6%),P0.05]。检测DM处理后细胞内活性氧水平研究发现,10、50μg/m L DM处理48 h 3株细胞均可见剂量依赖性ROS水平上升。微管蛋白免疫荧光染色可见采用IC50浓度的多西紫杉醇和10μg/m L DM处理48 h后,MKN-45荧光信号均表现为局部浓聚、紊乱。结论达玛-24-烯-3β-乙酰氧基-20s-醇能够将胃癌细胞周期阻滞于S期,诱导细胞凋亡,从而抑制细胞增殖作用。
[Abstract]:Objective to study the effect of alcohol extract of Forsythia suspensa on tumor cells in vitro. The antitumor effect of Dammar-24-ene-3 尾 -acetate-20S-olDMm extracted from Forsythia suspensa in vitro and its mechanism were further studied. Methods the inhibitory effect of DM on gastric cancer cell line SGC-7901BGC-823 and MKN-45 in vitro was detected by MTT assay. MKN-45 control group and its low level, In high dose group, BGC-823 control group and its low level, high dose group, high dose group, high dose group and control group were all fresh medium, low dose DM was 10 ~ 50 渭 g / mL, flow cytometry was used to detect the percentage of apoptotic cells, and the results were analyzed by Cell Quest software. The ratio of different cell cycle was recorded. The ROS level was detected by DCFH-DA probe. The experiment was divided into three groups: blank control group, docetaxel group and DM group. The microtubule aggregation reaction system was set up. The final concentration of docetaxel 10 渭 mol / L DM was 50 or 100 渭 mol / L, and the blank control group was not treated with drugs. The effects of DM on the microtubule system were studied by microtubule aggregation test and tubulin immunofluorescence staining. Results the inhibitory rates of 50 渭 g / mL on three gastric cancer cell lines were all above 80%. The IC50 was 11.72 卤1.35 渭 g / mL BGC-823N 17.19 卤0.82 渭 g / mL SGC-7901U 7.55 卤0.79 渭 g / mL DM for 48 h. The percentage of apoptotic cells in MKN-45 low dose group and high dose group [25.1 卤1.3% and 55.2 卤2.3%] was significantly higher than that in MKN-45 control group (13.2 卤2.4% vs 13.2 卤2.4%), and the percentage of apoptotic cells in control group was significantly lower than that in BGC-823 control group (13.2 卤2.4% vs 13.2 卤2.4%) after low density culture for 8 days, and the percentage of apoptotic cells in MKN-45 low dose group and high dose group [25.1 卤1.3% and 55.2 卤2.3%] was significantly higher than that in control group. The percentage of apoptotic cells in the high dose group [18.2 卤2.1%] was significantly higher than that in the control group [10.5 卤1.8%]. Compared with the control group of SGC-7901 [10.5 卤1.8%], the percentage of apoptotic cells in the SGC-7901 high dose DM group was increased [21.1 卤1.9%], but there was no significant difference between the high dose DM group and the SGC-7901 low-dose DM group (12.3 卤1.6%). The percentage of S phase cells in the low dose group of MKN-45 decreased [14.5 卤2.7)%vs(12.3 卤3.3% P0.05]. Compared with the control group of BGC-823, the percentage of S phase increased [12.2 卤5.4)%vs(20.2 卤2.1] in the low dose group of BGC-823. Compared with the control group of SGC-7901, the percentage of S phase cells in the low dose group of SGC-7901 increased [21.5 卤3.8)%vs(31.3 卤2.6%]. The results of oxygen level study showed that the increase of ROS level in dose-dependent manner was observed in 3 cells treated with 1050 渭 g / mL DM for 48 h. The fluorescence signals of MKN-45 were detected by IC50 concentration of docetaxel and 10 渭 g / mL DM for 48 h. Expressed as local concentration, Conclusion Dama 24 ene 3 尾 acetyl 20 s alcohol can block the cell cycle in S phase, induce apoptosis and inhibit the proliferation of gastric cancer cells.
【作者单位】：南京大学医学院附属鼓楼医院肿瘤中心;
【分类号】：R735.2

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