目的:研究抑制FA/BRCA途径中的范可尼贫血相关蛋白F(FANCF)基因在增加人肺腺癌CALU-1细胞株对顺铂敏感性中的作用。方法:设计靶向于FANCF基因的3条siRNAs(FANCF-siRNAs),用脂质体转染试剂将其分别转染于CALU-1肺癌细胞株,RT-PCR检测转染...目的:研究抑制FA/BRCA途径中的范可尼贫血相关蛋白F(FANCF)基因在增加人肺腺癌CALU-1细胞株对顺铂敏感性中的作用。方法:设计靶向于FANCF基因的3条siRNAs(FANCF-siRNAs),用脂质体转染试剂将其分别转染于CALU-1肺癌细胞株,RT-PCR检测转染后24 h FANCF mRNA的变化;筛选转染效率最高的siRNA片段。蛋白质印迹法检测经顺铂处理的CALU-1细胞株转染siRNA前后FANCF蛋白表达及FANCD2蛋白单泛素化水平;免疫荧光法测定胞核中FANCD2蛋白核聚小体的形成;CCK-8法测定转染siRNA前后的细胞增殖率变化。结果:与转染前比较,CALU-1肺癌细胞株转染FANCF-siRNA后FANCF蛋白表达明显下降,FANCD2蛋白单泛素化水平和核聚小体形成降低,细胞增殖率显著下降。结论:应用siRNA转染技术沉默FANCF基因可明显增加肺癌细胞对顺铂的敏感性,提示在肺癌靶向治疗策略中,FA/BRCA途径中的FANCF基因很可能是一个潜在的分子靶标。展开更多
Objective: To investigate the effects of xanthotoxin from Apiaceae medicinal plants on cell proliferation and apoptosis, and explore its mechanism of action against human gastric carcinoma SGC-7901 cells in vitro.Met...Objective: To investigate the effects of xanthotoxin from Apiaceae medicinal plants on cell proliferation and apoptosis, and explore its mechanism of action against human gastric carcinoma SGC-7901 cells in vitro.Methods: SGC-7901, HepG-2, MCF-7, and A549 cells were treated with different concentrations of xanthotoxin(10, 20, 60, 80, 100, 120, 140, and 160 μg/mL) for 48 h, and the cell viability(IC50) was determined by MTT assay; Xanthotoxin-induced apoptosis in cells was observed by using Hoechst 33258 Staining Kit and Annexin V-FITC Apoptosis Detection Kit; Flow cytometry was used to detect apoptosis related proteins of Fas/FasL, Bid, and DR5/TRAIL proteins in human gastric carcinoma SGC-7901 cells after being treated by xanthotoxin; The influence of xanthotoxin on Caspase-8 protein expression in the cells was determined by Flouormetric Assay Kit.Results: Xanthotoxin obviously inhibited SGC-7901, HepG-2, MCF-7, and A549 cells proliferation, and its inhibition was in a concentration-dependent manner; flow cytometry results showed that in a certain concentration range, xanthotoxin can increase the expression levels of Fas/FasL and DR5/TRAIL proteins in a concentration-dependence manner. The content of Bid protein in cells was increased, and it showed concentration-dependence.Conclusion: Xanthotoxin may induce SGC-7901 cells apoptosis in a certain concentration range through the Fas/FasL protein mediated death receptor pathway, or by DR5/TRAIL mediated death receptor pathway, and increase the expression level of death receptor protein, activation Caspase-8, activating downstream effect factor, inducing cell apoptosis, or activate Caspase-8 cutting activate protein Bid, and then enter the mitochondrial pathway, induction of apoptosis.展开更多
文摘目的:研究抑制FA/BRCA途径中的范可尼贫血相关蛋白F(FANCF)基因在增加人肺腺癌CALU-1细胞株对顺铂敏感性中的作用。方法:设计靶向于FANCF基因的3条siRNAs(FANCF-siRNAs),用脂质体转染试剂将其分别转染于CALU-1肺癌细胞株,RT-PCR检测转染后24 h FANCF mRNA的变化;筛选转染效率最高的siRNA片段。蛋白质印迹法检测经顺铂处理的CALU-1细胞株转染siRNA前后FANCF蛋白表达及FANCD2蛋白单泛素化水平;免疫荧光法测定胞核中FANCD2蛋白核聚小体的形成;CCK-8法测定转染siRNA前后的细胞增殖率变化。结果:与转染前比较,CALU-1肺癌细胞株转染FANCF-siRNA后FANCF蛋白表达明显下降,FANCD2蛋白单泛素化水平和核聚小体形成降低,细胞增殖率显著下降。结论:应用siRNA转染技术沉默FANCF基因可明显增加肺癌细胞对顺铂的敏感性,提示在肺癌靶向治疗策略中,FA/BRCA途径中的FANCF基因很可能是一个潜在的分子靶标。
基金supported by Harbin University of Commerce Research Funding(17x072)
文摘Objective: To investigate the effects of xanthotoxin from Apiaceae medicinal plants on cell proliferation and apoptosis, and explore its mechanism of action against human gastric carcinoma SGC-7901 cells in vitro.Methods: SGC-7901, HepG-2, MCF-7, and A549 cells were treated with different concentrations of xanthotoxin(10, 20, 60, 80, 100, 120, 140, and 160 μg/mL) for 48 h, and the cell viability(IC50) was determined by MTT assay; Xanthotoxin-induced apoptosis in cells was observed by using Hoechst 33258 Staining Kit and Annexin V-FITC Apoptosis Detection Kit; Flow cytometry was used to detect apoptosis related proteins of Fas/FasL, Bid, and DR5/TRAIL proteins in human gastric carcinoma SGC-7901 cells after being treated by xanthotoxin; The influence of xanthotoxin on Caspase-8 protein expression in the cells was determined by Flouormetric Assay Kit.Results: Xanthotoxin obviously inhibited SGC-7901, HepG-2, MCF-7, and A549 cells proliferation, and its inhibition was in a concentration-dependent manner; flow cytometry results showed that in a certain concentration range, xanthotoxin can increase the expression levels of Fas/FasL and DR5/TRAIL proteins in a concentration-dependence manner. The content of Bid protein in cells was increased, and it showed concentration-dependence.Conclusion: Xanthotoxin may induce SGC-7901 cells apoptosis in a certain concentration range through the Fas/FasL protein mediated death receptor pathway, or by DR5/TRAIL mediated death receptor pathway, and increase the expression level of death receptor protein, activation Caspase-8, activating downstream effect factor, inducing cell apoptosis, or activate Caspase-8 cutting activate protein Bid, and then enter the mitochondrial pathway, induction of apoptosis.