人肺腺癌细胞系中SIRT1表达与奈达铂敏感性的关系  

Expression of SIRT1 in human lung adenocarcinoma cells and its relation to the susceptibility of NDP

作　　者：毛旭华 陈姝颖 汤俊明 乔国洪 曹海霞[3] MAO Xuhua;CHEN Shuying;TANG Junming;QIAO Guohong;CAO Haixia(a.Department of Clinical Laboratory, 1b.Department of Pathology Laboratory, Yixing People＇s Hospital, Yixing 214200, Jiangsu;Research Center for Clinical Oncology , Jiangsu Cancer Hospital ＆ Jiangsu Institute of Cancer Research ＆ The Affiliated Cancer Hospital of Nanjing Medical Unlverslty,Nanjing 210009 Jiangsu,China)

机构地区：[1]江苏大学附属宜兴医院检验科,江苏无锡214200 [2]江苏大学附属宜兴医院病理科,江苏无锡214200 [3]江苏省肿瘤医院&江苏省肿瘤防治研究所&南京医科大学附属肿瘤医院临床肿瘤实验中心,南京210009

出　　处：《临床检验杂志》2018年第5期345-349,共5页Chinese Journal of Clinical Laboratory Science

基　　金：江苏省科技厅青年基金(BK20141016)

摘　　要：目的探讨5株人肺腺癌细胞系HCC827、H1650、H1975、A549和H1299中沉默信息调节因子1(Sirtuin 1,SIRT1)与奈达铂(Nedaplatin,NDP)药物敏感性的关系。方法采用实时定量PCR及western blot检测5株人肺腺癌细胞系SIRT1 m RNA及蛋白质表达水平;NDP作用于细胞后,用CCK-8法检测细胞活力并计算半数生长抑制浓度值(the half growth inhibition concentration,IC50);通过si RNA干扰技术下调A549、H1299、H1650和H1975细胞系中SIRT1的表达,采用CCK-8法和流式细胞术分别检测NDP对细胞存活率和细胞凋亡的影响。结果与HCC827细胞(m RNA和蛋白质相对表达量分别为1.00和0.11±0.02)相比,H1650、H1975、A549和H1299细胞中SIRT1 m RNA(分别为4.53±0.74、3.11±0.64、15.76±2.28和18.09±1.17)和蛋白质表达水平(分别为0.23±0.03、0.21±0.02、0.52±0.11和0.56±0.08)均明显上调,差异有统计学意义(F分别为122.10和26.50,P<0.01);A549和H1299细胞对NDP的敏感性[IC50值分别为(7.38±1.59)μmol/L和(8.14±1.43)μmol/L]明显高于HCC827、H1650和H1975细胞[IC50值分别为(26.16±4.35)μmol/L、(22.29±3.26)μmol/L和(24.41±2.58)μmol/L],差异有统计学意义(F=30.86,P<0.01)。A549和H1299细胞转染并经NDP处理后,si SIRT1组细胞存活率明显高于NC组(F分别为235.10和39.20,P<0.01),细胞凋亡率明显低于NC组(t分别为7.29和6.68,P<0.05);而在H1650和H1975细胞中,si SIRT1组细胞存活率明显低于NC组(F分别为185.40和60.09,P<0.01),细胞凋亡率明显高于NC组(t分别为6.15和31.36,P<0.01)。结论在SIRT1高表达的A549和H1299细胞中SIRT1表达增加了细胞对NDP敏感性,而SIRT1中表达的H1650和H1975细胞中SIRT1表达降低了细胞对NDP的敏感性,提示在人肺腺癌细胞中SIRT1可能在铂类耐药发挥双重作用。Objective To investigate the expression of Situin 1（ SIRT1） in 5 strains of human lung adenocarcinoma cell lines,including HCC827,H1650,H1975,A549 and H1299,and its relation to the susceptibility of nedaplatin（ NDP）. Methods The SIRT1 m RNA and protein levels in 5 strains of human lung adenocarcinoma cells were detected by real-time quantitative PCR and Western blot,respectively. The viability of cells treated with NDP was detected by the CCK-8 method and the half growth inhibition concentration（ IC50） was calculated. After the expressions of SIRT1 in A549,H1299,H1650 and H1975 cells were down-regulated by the si RNA interference,the effects of NDP on the viability and apoptosis of these cells were determined by the CCK-8 method and flow cytometry,respectively.Results The expression levels of SIRT1 m RNA（ 4.53 ± 0.74,3.11 ± 0.64,15.76 ± 2.28 and 18.09 ± 1.17） and protein（ 0.23 ± 0.03,0.21 ± 0.02,0.52 ± 0.11 and 0.56 ± 0.08） in H1650,H1975,A549 and H1299 cells were significantly higher than that in HCC827 cells（ 1.00 for SIRT1 m RNA and 0.11 ± 0.02 for SIRT1 protein,F = 122.10 and 26.50,respectively,P〈0.01）.The susceptibility of A549 and H1299 cells to NDP [IC50=（ 7.38 ± 1.59） and（ 8.14 ± 1.43） μmol/L,respectively]was significantly higher than that of HCC827,H1650 and H1975 cells [IC50=（ 26.16±4.35）,（ 22.29±3.26） and（ 24.41 ± 2.58）,respectively,F =30.86,P〈0.01].The survivals of A549 and H1299 cells transfected by si SIRT1 and treated with NDP were significantly higher than that in the NC group（ F = 235.10 and 39.20,respectively,P〈0.01）,and the apoptotic rates were the reverse（ t = 7.29 and 6.68,respectively,P〈0.05）. However,the survivals of H1650 and H1975 cells transfected by si SIRT1 and treated with NDP were significantly lower than that in the NC group（ F = 185.40 and 60.09,respectively,P〈0.01）,and the apoptotic rates were the reverse（ t = 6.15 and31.36,respectively,P〈0.01）.Conclusion The expression of SIRT1 in A549 and H129

关 键 词：肺癌 沉默信息调节因子1 奈达铂 

分 类 号：R734.2[医药卫生—肿瘤]