丝石竹皂甙对人胃癌细胞SGC-7901增殖、凋亡的影响及机制研究

发布时间：2018-03-11 15:13

  本文选题：丝石竹皂甙　切入点：胃癌　出处：《青岛大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:探究丝石竹提取物丝石竹皂甙在体外对人胃腺癌细胞株SGC-7901增殖、凋亡的影响及其机制。方法:取对数生长期的SGC-7901细胞作为研究对象,实验组加入不同浓度(20、40、80ug/ml)的丝石竹皂甙培养液,以不加入丝石竹皂甙仅加入等体积培养基(含10%胎牛血清)的细胞作为对照组,分别培养24、48、72h。MTT法检测细胞增殖活性;Annexin V/PI双染法结合流式细胞仪分析细胞凋亡;流式细胞术DNA含量分析法分析细胞周期;实时荧光定量PCR检测Wnt5a、Wnt8b基因的表达。结果:(1)①不同浓度的丝石竹皂甙作用相同时间,细胞存活率的差异具有统计学意义(p0.01)。24h细胞存活率:对照组为100.00%,实验组(按浓度由低到高排列,下同)依次为(82.82±1.64)%、(50.58±1.04)%、(20.00±0.75)%,F=846.157;48h细胞存活率:对照组为100.00%,实验组依次为(76.65±1.48)%、(40.28±1.40)%、(16.38±0.88)%,F=896.265;72h细胞存活率:对照组为100.00%,实验组依次为(70.57±1.60)%、(27.47±1.21)%、(10.09±0.45)%,F=957.040。②相同浓度丝石竹皂甙作用不同时间(对照组除外),细胞存活率的差异具有统计学意义。当浓度为20ug/ml时,F=45.281,p0.01;浓度为40ug/ml时,F=212.674,p0.01;浓度为80ug/ml时,F=129.148,p0.01。(3)计算求得24小时IC50值为41.171ug/ml。(2)不同浓度的丝石竹皂甙作用24h后应用流式细胞术AV/PI双染法进行细胞凋亡率分析:对照组为(15.11±0.95)%,实验组依次为(20.33±0.78)%、(23.93±1.18)%、(27.67±1.44)%,F=83.215,p0.01。(3)不同浓度的丝石竹皂甙作用24h后应用流式细胞术DNA含量分析法进行细胞周期分析,结果以百分比表示,G0/G1期:对照组为(53.99±1.35)%,实验组依次为(58.40±1.77)%、(62.40±0.85)%、(65.43±1.18)%,F=42.792,p0.01;S期:对照组为(31.64±1.30)%,实验组依次为(27.07±1.38)%、(23.11±1.47)%、(19.81±0.70)%,F=53.396,p0.01。G2期:对照组为(14.71±0.06)%、实验组依次为(14.91±0.18)%、(14.67±007)%、(14.71±0.17)%,F=2.209,p0.05。(4)不同浓度的丝石竹皂甙作用48h后应用实时荧光定量PCR进行基因表达分析,基因表达结果以相对比表示,Wnt5a/GAPDH1组:对照组为1.70±0.09,实验组依次为1.22±0.10、0.74±0.13、0.58±0.07,F=109.516,p0.01;Wnt8b/GAPDH1组:对照组为1.18±0.08,实验组依次为0.82±0.07、0.53±0.06、0.42±0.03,F=104.298,p0.01。结论:丝石竹皂甙可能通过下调Wnt基因的表达,在体外抑制胃癌细胞增殖、促进其凋亡,并使其细胞周期阻滞于G0/G1期,减少S期细胞比例。
[Abstract]:Objective: To explore the Gypsophila extract gypsoside in vitro on human gastric cancer cell SGC-7901 proliferation, apoptosis and its mechanism. Methods: SGC-7901 cells in logarithmic growth phase as the research object, different concentration in experimental group (20,40,80ug/ml) of gypsoside medium, to join gypsoside only adding volume (culture medium containing 10% fetal bovine serum) cells were cultured as the control group, 24,48,72h.MTT was used to detect the proliferation activity; analysis of apoptotic cells by flow cytometry with Annexin V/PI double staining method; flow cytometry DNA content analysis method to analyze the cell cycle; real time fluorescence quantitative PCR detection of Wnt5a, Wnt8b gene expression. Results: (1) the same time gypsoside effect of different concentration, the difference was statistically significant cell survival rate (P0.01):.24h cell survival was 100% in the control group, the experimental group (according to concentration by 浣庡埌楂樻帓鍒，

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