达玛-24-烯-3β-乙酰氧基-20s-醇对胃癌细胞增殖的抑制作用  

Anti-proliferation effects of dammar-24-ene-3β-acetate-20S-ol on gastric cancer cells

作　　者：谢丽[1] 胡文静[1] 朱丽晶[1] 刘宝瑞[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肿瘤中心,南京医学博士210008

出　　处：《医学研究生学报》2017年第5期481-485,共5页Journal of Medical Postgraduates

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(P<0.01)。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(P<0.01)。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(P<0.05),而与SGC-7901低剂量DM组[(12.3±1.6)%]差异无统计学意义(P>0.05)。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)%,P>0.05]。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)%,P<0.05]。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%)Objective Based on the previous research that the ethanolic extract from traditional Chinese medicine fructus forsythiae（ Lianqiao） can obviously inhibit cancer cells in vitro,the article aimed to investigate the anti-proliferation effects of dammar-24-ene-3β-acetate-20S-ol（ DM） extracted from fructus forsythiae on gastric cancer cells and its mechanism. Methods MTT assay was used to assess the anti-proliferation effects of DM on gastric cancer cells including SGC-7901,BGC-823,and MKN-45 in vitro. There were MKN-45 control group and its low dose and high dose groups,BGC-823 control group and its low dose and high dose groups,SGC-7901 control group and its low dose and high dose groups in the experiment. Flow cytometry was used to analyze the cell apoptosis rate.Cellquest software was used to analyze the results and record the ratio of cells at different cycles. DCFH-DA probe was applied to detect the ROS levels of blank control group,docetaxol group and DM group. The reaction system of microtubule assembly test was set with10？ mol/L docetaxol,50 or 100 μmol/L DM final density and no medicine in blank control group. The readings of UV spectrophotometer were recorded. Microtubule assembly assay and microtubule immunofluorescence staining were applied to investigate the effects of DM on microtubule system. Results The inhibition ratio of 50 μg/L DM on the proliferation three gastric cell lines were all above80%,with IC50 s of 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. 8 days’ low density culturing at 48 hours after 2 μg/m L DM treatment,compared with control group,the number of cell clones significantly reduced without much change in clone size,while 48 hours after 10 μg/m L DM treatment,besides a few clones of BGC-823,there were just several megascopic clones of SGC-7901 and MKN-45. In comparison with apoptotic cell ratio in MKN-45 control group[（ 21.1± 2.5） %],its low dose group and high dose group resulted in significant rise of apoptotic cell ratio[

关 键 词：达玛-24-烯-3β-乙酰氧基-20s-醇 胃癌细胞 增殖抑制 

分 类 号：R735.2[医药卫生—肿瘤]