【作者】 张小芳；

【导师】 赵健雄；

【作者基本信息】 兰州大学 ， 中西医结合临床， 2010， 硕士

【摘要】 目的：研究葡萄籽原花青素对肺腺癌SPC-A-1细胞X射线放疗的增敏作用。方法：1.应用MTT法和集落形成法研究原花青素的放疗增敏效应,单因素方差分析确定增敏剂量和增敏作用,单击多靶曲线拟合计算增敏比及相关参数。2.流式细胞仪检测空白对照组(0Gy),单独X射线照射组(1Gy,5Gy)和药物增敏X射线组(100μg·mL-1原花青素1Gy,5Gy)细胞周期变化与凋亡3.荧光定量PCR检测单独X射线照射组(1Gy)和不同浓度药物增敏组(50μg·mL-1,100μg·mL-1,1Gy)对细胞周期调控因子cyclin D1表达的影响。结果：1.MTT检测结果表明,单独应用原花青素对SPC-A-1的抑制作用较弱,IC50大于400 ug/ml;细胞药物处理后对X射线的敏感性增加,单因素方差分析表明药物和X射线有协同作用,50 ug/ml时表现出增敏效应；集落形成结果表明,100 ug/ml药物对X射线增敏作用明显,增敏比为2.56。2.流式细胞术检测发现试验组细胞出现了明显的凋亡峰提示亚二倍体DNA含量的细胞数量明显增加,细胞凋亡率(28.4%)显著高于单独照射组(14.9%)及空白对照组(5.3%)。处理组与相应对照组分别比较差异有非常显著性意义(p<0.01)。实验结果说明,原花青素可以诱导细胞凋亡,并将细胞阻滞于G1期。3.荧光定量PCR结果发现,原花青素50、100μg·mL-1作用于细胞48 h较对照组CyclinD1mRNA的表达分别下降0.189和0.426倍,表明原花青素可下调细胞CyclinD1mRNA的转录表达。结论：葡萄籽原花青素对肺腺癌SPC-A-1细胞有显著的放疗增敏作用。其机制可能是通过抑制细胞的增殖,G1期阻滞,诱导细胞凋亡,下调CyclinD1基因表达等而发挥抗肿瘤作用。更多还原

【Abstract】 Objective:To study the enhancement radiosensition of grape seed procyanidin extract for X-ray radiotherapy on lung cancer cell line SPC-A-1 in vitro and try to explore its mechanism.Methods:1.The enhancement of radiosensitising effect was detected by clone formation experiment and MTT methods. The radiosensitising dose of anthocyanidin was determined by single-factor analysis of variance. The ratio of sensitization and relevant parameters were calculated by the multi-target/single-hit model.2.Flow cytometry (FCM)was used to measure cell cycle and apoptosis in radiotherapy group and increasing radiatesensitivity group.3.Compared with control group, the expression of CyclinDl mRNA of a variety of concentration of the radiosensitising group analyzed by FPCR.Results:1.The results of MTT indicated that the inhibition effect on SPC-A-1 was slight when only using procyanidin, and the IC50 greater than 400 ug/ml. Though, after the lung cancer cell line was administrated by procyanidin, the sensitivity of X-ray radiotherapy was enhanced. The study showed that the procyanidin and X-ray has synergistic effects, the dose of 100 ug/ml has the significant sensitizing effect, and the ratio of sensitization was 2.56.2.Flow cytometry analysis showed that procyanidin induced apoptosis of SPC-A-1 cells (The apoptotic rate were 28.4%,14.9%,5.3%, respectively.)and inhibited the proceeding of cell cycle in Gl period.3.PCR showed that the CyclinD1 protein expression were down-regulated by administration of procyanidin.Conclusion:Grape seed procyanidin extract has the significant sensitizing effect on radiotherapy in lung adenocarcinoma cells SPC-A-1. GSEP inhibits the proliferation of malignant human Lung adenocarcinoma cancer cells and induces their apoptosis. Expression of CyclinD1 protein may be related to cell growth inhibition and the apoptosis mediated by GSEP in vitro.更多还原