Xie Ling-Xia, Hu Li-Na, Chen Heng-Xi, Peng Xue, Li Jin-Ke
Department of Gynecology and Obstetrics, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2013 Jan;44(1):36-41.
To observe whether cisplatin-induced apoptosis were increased when SiHa cells were preincubated with nuclear factor-kappa B (NF-kappaB) inhibitors [aspirin, sulindac, curcumin or pyrrolidine dithiocarbamate (PDTC)].
SiHa cells 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.
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-kappaB 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.
Aspirin, sulindac, curcumin and PDTC could all inhibit cisplatin induced NF-kappaB activiation, which could increase cispaltin-induced chemosensativity by augments of apoptosis.
观察当SiHa细胞与核因子-κB(NF-κB)抑制剂[阿司匹林、舒林酸、姜黄素或吡咯烷二硫代氨基甲酸盐(PDTC)]预孵育时,顺铂诱导的细胞凋亡是否增加。
将SiHa细胞分别与阿司匹林、舒林酸、姜黄素和PDTC预孵育2小时,然后再与顺铂孵育,采用蛋白质免疫印迹法检测细胞核提取物中P65的水平。采用MTT法检测相对细胞活力。采用TUNEL法检测凋亡率。还用PI染色的流式细胞术检测细胞凋亡及细胞周期。
当SiHa细胞用阿司匹林、舒林酸、姜黄素或PDTC预处理时,蛋白质免疫印迹显示,在顺铂刺激下P65的表达受到抑制(P<0.05)。MTT法表明,用NF-κB抑制剂预孵育可显著增加顺铂诱导的化学敏感性(P<0.05)。当细胞用阿司匹林、舒林酸、姜黄素或PDTC预处理时,TUNEL法和流式细胞术检测显示,顺铂刺激24小时后凋亡率均增加(P<0.05)。流式细胞术的结果还显示,用阿司匹林、舒林酸、姜黄素或PDTC预处理可显著增加顺铂诱导的细胞凋亡。
阿司匹林、舒林酸、姜黄素和PDTC均可抑制顺铂诱导的NF-κB激活,通过增加细胞凋亡来提高顺铂诱导的化学敏感性。