核转录因子-κB组成性激活对胃癌细胞增殖的影响及其机制  

Effect of the constitutive activation of NF-κB on the proliferation of gastric cancer cells

作　　者：李琦[1] 于颖彦[1] 朱正纲[1] 刘炳亚[1] 纪玉宝[1] 张奕[1] 程琳玲 林言箴[1] 

机构地区：[1]上海第二医科大学附属瑞金医院上海消化外科研究所,200025 [2]上海免疫学研究所

出　　处：《中华实验外科杂志》2004年第8期957-960,共4页Chinese Journal of Experimental Surgery

摘　　要：目的 探讨胃癌细胞株是否存在核转录因子(NF)-κB组成性激活及其对胃癌细胞增殖的影响机制。方法 采用蛋白免疫印迹法(Western blot法)检测4株不同分化程度的胃癌细胞株NF-κB蛋白表达,比较4株胃癌细胞株中NF-κB蛋白质表达水平的差异。通过Trans AM^(TM)NFκBp65试剂盒来比较不同胃癌细胞株中p65亚基的蛋白活性差异。选取活性较高细胞株,观察吡咯烷二硫代氨基甲酸盐(PDTC)对NF-κB活性的影响;利用噻唑蓝(MTT)法,分别观察24、48、72h,PDTC对细胞增殖的影响。并采用流式细胞分析技术(FCM)检测细胞的凋亡情况。结果 4株胃癌细胞株胞核中均存在NF-κB蛋白的表达,即存在NF-κB的组成性激活,且存在表达差异。活化的p50亚基在AGS细胞株中表达较低,在MKN28、MKN45、SGC-7901细胞株中表达较高;活化的p65亚基在MKN28、MKN45细胞株中表达较低,在AGS、SGC-7901细胞株中表达较高。经PDTC处理的实验组胃癌细胞,NF-κB的活性均被抑制(P<0.01);同时细胞表现为不同程度的增长抑制(P<0.01),FCM结果显示PDTC可诱导细胞的凋亡。结论 胃癌细胞株中存在NF-κB的组成性激活,并在不同的细胞株中存在表达差异。抑制NF-κB的活性可明显抑制胃癌细胞的增殖,其机制与PDTC所诱导的细胞凋亡有关。Objective To study if there exists the constitutive activation of NF-κB and its effect on gastric cancer strains cell proliferation. Methods The different expression of NF-κB was detected by Western blot among 4 different differential gastric cancer strains. Trans AM^(TM) NF-κB p65 Kit was used to detect and compare the difference of the activity of p65. The effect of PDTC on the gastric cancer cell proliferation was detected by MTT at different time points. FCM was used to detect the apoptosis of the cells. Results In the 4 gastric cancer strains, there existed the constitutive activation of NF-κB. The expression of the activated subunit of p50 was lower in AGS cell strain, but higher in MKN28, MKN45, SGC-7901 cell strains. The expression of the activated subunit of p65 was lower in MKN28 and MKN45 cell strains, while higher in AGS and SGC-7901 cell strains. The activity of subunit p65 was the highest in SGC-7901 cell strain. The activity of NF-κB and the cell proliferation was inhibited in experimental groups treated with PDTC, as compared with control groups ( P<0.01 ). PDTC could induce cell apoptosis. Conclusion There exists the constitutive activation of NF-κB and different expression of NF-κB protein in gastric cancer strains. Inhibiting the activity of NF-κB can inhibit the cell proliferation, and it may be related to cell apoptosis induced by PDTC.

关 键 词：NF-κB 胃癌细胞株 PDTC 表达差异 胃癌细胞增殖 激活 MKN45 蛋白活性 蛋白免疫印迹法 蛋白质表达 

分 类 号：R735.2[医药卫生—肿瘤]