PDTC对胃癌细胞MKN-45 p53异构体表达的影响及其生物学意义  

Effect of PDTC on Expression of p53 Isoforms in Gastric Cancer Cell Line MKN-45 and Its Biological Significance

作　　者：王燕泽 崔璨[1] 张红梅[2] 李蕾[2] 张丽[2] 王春芽 焦建新[2] 高志星[2] 季万胜[2] 

机构地区：[1]潍坊医学院,山东潍坊261053 [2]潍坊医学院附属医院,山东潍坊261031

出　　处：《中国肿瘤》2017年第2期156-160,共5页China Cancer

基　　金：山东省优秀中青年科学家科研奖励基金(BS2010SW034)

摘　　要：[目的]探讨核因子-κB(NF-κB)抑制剂吡咯烷二硫代氨基甲酸(PDTC)对胃癌细胞MKN-45 p53异构体表达的影响及其生物学意义。[方法 ]不同浓度的PDTC(25、50、75μmol/L)单独及联合顺铂(4μg/ml)作用于胃癌细胞MKN-45,采用细胞增殖/毒性检测试剂盒(CCK-8试剂盒)检测细胞增殖抑制率;实时荧光定量多聚酶链反应(RT-PCR)检测p53β、Δ133p53及NF-κB p65在m RNA水平的表达情况。[结果 ]CCK-8结果显示,PDTC(25、50、75μmol/L)单独作用时能抑制MKN-45细胞生长,抑制率分别为4.95%、12.20%、21.28%,差异有统计学意义(P<0.0001);当与顺铂(4μg/ml)联合时,MKN-45细胞增殖抑制率随PDTC浓度的增加而增加,且高于单独顺铂组,差异有统计学意义(P<0.001)。RT-PCR结果显示,当用不同浓度PDTC(25、50、75μmol/L)预处理2h后再加入顺铂(4μg/ml),MKN-45细胞中p53βm RNA的表达差异无统计学意义(P=0.04),而Δ133p53和p65 m RNA的表达随PDTC浓度的增加而降低,且低于顺铂单独组,差异有统计学意义(P<0.0001)。Pearson相关性分析显示,p65与p53βm RNA表达无相关性(r=0.072,P=0.798),p65与Δ133p53 m RNA表达呈正相关(r=0.814,P<0.0001)。[结论 ]NF-κB抑制剂PDTC可以抑制MKN-45细胞的生长,也可以增强顺铂对该细胞的生长抑制效应,Δ133p53异构体可能是NF-κB-p53对话的关键分子。[Purpose] To investigate the effect of the nuclear factor-κB（NF-κB） inhibitor pyrrolidine dithiocarbamate（PDTC） on the expression of p53 isoforms in gastric cancer MKN-45 cells and its biological significance. [Methods] Different concentrations of PDTC （25,50,75μmol/L） alone or in combination with cisplatin （4μg/ml） were used to treat gastric cancer MKN-45 cells. The inhibition rate of MKN-45 cells was determined by cell proliferation/toxicity testing kits（CCK-8 assay）. The mRNA expressions of p53β,Δ133p53 and NF-κB p65 were determined by real-time polymerase chain reaction（RT-PCR）. [Results] CCK-8 assay showed that PDTC（25,50,75μmol/L） inhibited the growth of gastric cancer MKN-45 cells with an inhibition rate of 4.95%,12.20% and 21.28% respectively（P〈0.0001）. When combined with cisplatin,the inhibition rate of PDTC was higher than PDCT alone（P〈0.001）. RT-PCR showed that,when cisplatin（4μg/ml） was added in PDTC-pretreated MKN-45 cells,the expressions of Δ133p53 and p65 mRNA were reduced and lower than that in cells treated by cisplatin alone（P〈0.0001）;however,there were no significant changes of p53β mRNA expression（P=0.04）. Pearson correlation analysis showed that there was no correlation between mRNA expression of p65 and p53β（r=0.072,P=0.798）.The mRNA expression of p65 was positively correlated with Δ133p53（r=0.814,P〈0.0001）. [Conclusion] NF-κB inhibitor PDTC can inhibit the growth of gastric cancer MKN-45 cells,and enhance inhibitory effect of cisplatin. The results indicate that Δ133p53 rather than p53β isoforms might be a key molecule of the NF-κB-p53 interaction.

关 键 词：p53β Δ133p53 NF-ΚB P65 胃癌 吡咯烷二硫代氨基甲酸 顺铂 

分 类 号：R735.9[医药卫生—肿瘤]