核转录因子-κB抑制剂咖啡酸苯乙基酯对肾癌细胞株786-O侵袭力和凋亡的影响  

Influence of nuclear factor-κB inhibitor caffeic acid phenethyl ester on the invasiveness and apoptosis of renal carcinoma cell line786-O

作　　者：王波[1] 王光春[1] 郑军华[1] 

机构地区：[1]同济大学附属第十人民医院泌尿外科,上海200072

出　　处：《上海医学》2013年第7期605-608,共4页Shanghai Medical Journal

基　　金：国家自然科学基金(30972984);上海市市级医院新兴前沿技术联合攻关项目(SHDC12010104);上海市卫生局新百人计划(XBR2011021)

摘　　要：目的探讨核转录因子(NF)-κB特异性抑制剂咖啡酸苯乙基酯(CAPE)对肾透明细胞癌细胞株786-O侵袭力和凋亡的影响。方法对照组以0.1%二甲基亚砜(DMSO)孵育786-O细胞株24h,CAPE组以10μmol/L CAPE孵育786-O细胞株24h后,采用实时定量聚合酶链反应检测相关基因mRNA。分别以1μmol/L CAPE(CAPE 1μmol/L组)和10μmol/L CAPE(CAPE 10μmol/L组)孵育786-O细胞株24h,采用Western印迹法检测相关蛋白的表达水平。应用流式细胞仪分析细胞凋亡和周期的改变。结果 CAPE组的NF-κB、转录活化因子3(STAT3)、凋亡相关基因B细胞淋巴瘤(Bcl)-2和Bcl-xl、肿瘤迁移相关基因基质金属蛋白酶(MMP)2和MMP9的mRNA相对表达量分别为90.212±0.141、0.339±0.172、0.034±0.006、0.332±0.070、0.028±0.004、0.006±0.001,均显著低于对照组的1(P值均<0.01)。CAPE 10μmol/L组的NF-κB、磷酸化NF-κB(p-NF-κB)、STAT3和磷酸化STAT 3(p-STAT 3)的蛋白表达量均显著低于对照组(P值均<0.05),CAPE 10μmol/L组与CAPE 1μmol/L组间的差异均无统计学意义(P值均>0.05)。CAPE 1μmol/L组、CAPE 10μmol/L组在S期的细胞数均显著少于对照组(P值均<0.05),在G0/G1期的细胞数均显著多于对照组(P值均<0.05)。对照组的细胞凋亡率为(1.97±1.11)%,显著低于CAPE 1μmol/L组的(11.13±4.31)%和CAPE 10μmol/L组的(52.00±15.62)%(P值均<0.05)。结论 CAPE能有效抑制786-O肾癌细胞株的迁移与增殖,并促进其凋亡,其机制可能是抑制了NF-κB的激活,进而下调其下游的相关基因。CAPE可有效地降低STAT3mRNA和蛋白水平的表达,NF-κB与STAT3在肾癌中可能存在相互作用。Objective To investigate the influence of caffeic acid phenethyl este（CAPE）,a nuclear factor-κB（NF-κB）inhibitor,on the invasiveness and apoptosis of renal carcinoma cell line 786-O.Methods 786O cells were incubated in culture media containing 0.1%dimethyl sulfoxide（DMSO）for 24hin control group while in 10μmol/L CAPE for 24hin CAPE group.The mRNA levels of related genes were detected by real-time quantitative polymerase chain reaction（RT-PCR）.Meanwhile,786-O cells were incubated in culture media containing 1μmol/L and 10μmol/L CAPE respectively to detect the expression of related proteins by western blot.Apoptosis and cell cycle were detected by flow cytometry.Results The mRNA expression of NF-κB,signal transducer and activator of transcription 3（STAT3）,B-cell lymphoma 2（Bcl-2）,Bcl-xl,matrix metallopeptidase 2（MMP2）and MMP9in the CAPE group were 90.212±0.141,0.339±0.172,0.034±0.006,0.332±0.070,0.028±0.004,and 0.006±0.001,respectively,which were significantly decreased compared with the control group（all P0.01）.The protein levels of NF-κB,p-NF-κB,STAT3and p-STAT3in the CAPE 10μmol/L group were significantly lower than those in the control group（all P0.05）,but there was no significant difference between the 10μmol/L and 1μmol/L groups（all P0.05）.Flow cytometry analysis showed that the S phase cell numbers in both the 1μmol/L and 10μmol/L CAPE groups were significantly less than the control group（all P 0.05）,while the G0/G1phase cell number was significantly more than the control group（all P0.05）.The apoptosis rate in the control group（[1.97±1.11]%）was significantly lower than that in the 1μmol/L CAPE group（[11.13±4.31]%）and 10μmol/L CAPE group（[52.00±15.62]%,all P0.05）.Conclusion CAPE can effectively inhibit the migration and proliferation of renal carcinoma cell line 786-O and promote its apoptosis,which may be due to the inhibition of NF-κB activation and subsequent down-regulation of its downstream genes.Meanwhile,CAPE can down-r

关 键 词：核转录因子ΚB 咖啡酸苯乙基酯 肾细胞癌 

分 类 号：R737.11[医药卫生—肿瘤]