NF-κB抑制物对顺铂诱导宫颈癌SiHa细胞凋亡的影响  

Effects of NF-κB Inhibitor on Cisplatin-induced Apoptosis in Cervical Cancer

作　　者：谢灵遐[1] 胡丽娜[2] 陈恒禧[1] 彭雪[1] 李金科[1] 

机构地区：[1]四川大学华西第二医院妇产科,成都610041 [2]重庆医科大学附属第二医院妇产科,重庆400010

出　　处：《四川大学学报（医学版）》2013年第1期36-41,共6页Journal of Sichuan University(Medical Sciences)

基　　金：中央高校基本科研业务费科研基地引导专项子课题基金(No.2010SCU3003)资助

摘　　要：目的了解核转录因子-κB(NF-κB)抑制物〔阿司匹林、舒林酸、姜黄和二硫氨基甲酸酞吡咯烷(PDTC)〕对顺铂介导的SiHa细胞凋亡的影响。方法体外培养宫颈癌SiHa细胞株,用前述4种NF-κB抑制物预处理细胞后,加入顺铂继续培养24h,Western blot检测NF-κB P65的激活情况;MTT检测细胞的相对存活率;采用原位缺口末端标记法(TUNEL)比较4种药物联合顺铂和单用顺铂处理24h细胞的凋亡;采用PI染色-流式细胞术检测细胞凋亡和细胞周期的改变。结果 NF-κB抑制物预处理后,再加入顺铂刺激,Western blot结果显示可抑制顺铂诱导的P65表达(P<0.05);MTT结果显示NF-κB抑制物能增加不同浓度顺铂对SiHa细胞的化疗毒性(P<0.05);NF-κB抑制物预处理后加入顺铂,TUNEL和流式细胞术检测细胞凋亡率均增加(P<0.05);顺铂能使S期细胞数增加(22.7%vs.31.2%,P<0.05),而NF-κB抑制物预处理细胞则能抑制S期细胞的数量。结论NF-κB抑制物能增加SiHa细胞对顺铂化疗的敏感性,其机制可能是通过抑制小剂量顺铂对SiHa细胞NF-κB的激活,同时抑制小剂量顺铂对细胞DNA合成的诱导作用,进而增加顺铂诱导的细胞凋亡而达到的。Objective To observe whether cisplatin-induced apoptosis were increased when SiHa cells were preincubated with nuclear factor-kappa B （NF-κB） inhibitors [aspirin, sulindac, curcumin or pyrrolidine dithiocarbamate （PDTC）]. Methods SiHa ceils were preincubated 2 hours with aspirin, sulindac, curcumin and PDTC respectively, then a further incubation were done with cisplatin, and Western blot analysis were applied to detect P65 level of nuclear extraction. MTT assay was done to detect relative cell viability. TUNEL was applied to detect apoptosis rates. Flow cytometryies with PI staining were also used to detect apoptosis as well as cell cycle. Results When SiHa cells were pretreated with aspirin, sulindac, curcumin or PDTC, Western blot showed that the expression of P65 was inhibited upon cisplatin stimulus （P〈0.05）. MTT assay demonstrated that a preincubation with NF-κB inhibitor could signifianctly increase cisplatin-induced chemosensitivity （P 〈 0. 05）. When cells pretreated with aspirin, sulindac, curcumin, or PDTC, TUNEL and flow cytometries assay showed that the apoptotic rates were all increased after 24 hours cisplatin stimulus （P〈0.05）. Results of flow cytometries were also showed that a pretreation with aspirin, sulindac, curcumin, or PDTC could significantly increase cisplatin-induced apoptosis. Conclusion Aspirin, sulindac, curcumin and PDTC could all inhibit cisplatin induced NF-κB activiation, which could increase cispaltin-induced chemosensativity by augments of apoptosis.

关 键 词：核转因子录-κB 宫颈癌 顺铂 细胞凋亡 化疗 

分 类 号：R737.33[医药卫生—肿瘤]