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沉默干扰素调控因子2降低胰腺癌细胞有氧糖酵解水平的实验研究 预览

Silencing IRF-2 reduces the level of aerobic glycolysis in pancreatic cancer cells
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摘要 目的:研究干扰素调控因子2(interferon regulatory factor 2,IRF-2)对胰腺癌细胞有氧糖酵解水平的影响。方法:胰腺癌细胞感染含有IRF-2-shRNA的慢病毒及对照空载体病毒记为干扰组和阴性组,以不做处理的细胞作为对照组,用Realtime PCR和Western blot方法测定细胞中IRF-2水平,MTT方法测定胰腺癌细胞的增殖情况,Western blot方法测定细胞内糖酵解关键酶己糖激酶2(HK2)、丙酮酸激酶M2亚型(PKM2)蛋白、NF-κBp65(NF-κBp65亚型)、细胞核增殖抗原(Ki-67)表达水平,用试剂盒检测细胞乳酸生成及葡萄糖消耗水平,同时检测细胞中三磷酸腺苷(ATP)合成水平。结果:干扰组细胞中的IRF-2 mRNA和蛋白水平均明显低于对照组,而阴性组细胞中IRF-2 mRNA和蛋白水平与对照组比较没有明显差异。干扰组细胞的OD值明显降低,细胞中HK2、PKM2、NF-κBp65、Ki-67蛋白水平降低,同时细胞乳酸生成及葡萄糖消耗量均降低,细胞合成的ATP减少,与对照组相比,差异有统计学意义(P<0.05)。阴性组细胞OD值、HK2蛋白水平、PKM2蛋白水平、Ki-67蛋白水平、NF-κBp65蛋白水平、乳酸生成、葡萄糖消耗量、ATP水平与对照组相比,差异均没有统计学意义(P> 0. 05)。结论:沉默IRF-2抑制胰腺癌细胞中糖酵解关键酶表达,降低细胞糖酵解水平,抑制胰腺癌细胞增殖。 Objective:To study the effect of IRF-2 on aerobic glycolysis in pancreatic cancer cells.Methods:Pancreatic cancer cells infected with lentivirus containing IRF-2-shRNA and control empty virus were classified as interference group and negative group.The cells not treated were used as the control group.The IRF-2 levels in cells were measured by Realtime PCR and Western blot.MTT assay was used to determine the proliferation of pancreatic cancer cells.Western blot method was used to detect the expression levels of HK2,PKM2,NF-κBp65,Ki-67 proteins in glycolysis.The cell lactate production and glucose consumption were detected by kit.At the same time,the level of ATP synthesis in the cells was also detected.Results:The transcriptional and protein levels of IRF-2 in the interference group were significantly lower than those in the control group.The IRF-2 transcription and protein levels in the negative group were not significantly different from those in the control group.The OD values of the cells in the interference group decreased significantly,the level of HK2,PKM2,NF-κBp65,Ki-67 protein in the cells decreased,at the same time,lactic acid generation and glucose consumption decreased,ATP decreased in cell synthesis.Compared with the control group,the difference was statistically significant(P<0.05).The OD value,HK2 protein level,PKM2 protein level,NF-κBp65 protein level,Ki-67 protein level,lactate production,glucose consumption and ATP level in the negative group were compared with those in the control group,and the difference was not statistically significant(P>0.05).Conclusion:Silence IRF-2 inhibits the expression of key glycolysis enzymes in pancreatic cancer cells,reduces the level of glycolysis and inhibits the proliferation of pancreatic cancer cells.
作者 刘婉 冯世兵 冯晓洁 王媛媛 Liu Wan;Feng Shibing;Feng Xiaojie;Wang Yuanyuan(Department of Digestive,Heping Li Hospital,Beijing 100013,China)
出处 《现代肿瘤医学》 CAS 2019年第14期2463-2468,共6页 Modern Oncology
关键词 胰腺癌 干扰素调控因子2 糖酵解 三磷酸腺苷 pancreatic cancer interferon regulatory factor 2 glycolysis adenosine triphosphate
作者简介 刘婉(1980-),女,江西人,硕士,主治医师,研究方向:消化系统肿瘤诊疗。
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