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[目的]探讨NF-κB抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)增敏卡铂对卵巢癌细胞的毒性作用及其机制。[方法]采用凝胶迁移率变动试验(EMSA)法,观察卡铂作用下和卡铂与PDTC共同作用下NF-κB的活化反应;MTT和流式细胞仪分别比较2种情况下SKOV3细胞的生长情况和凋亡率;RT-PCR方法检测2种情况下survivinmRNA的表达。[结果]EMSA实验结果表明,卡铂可以诱导NF-κB活化,PDTC可以抑制此作用。PDTC预处理后可增强卡铂对卵巢癌SKOV3细胞的生长抑制作用。卡铂作用24h、48h后卵巢癌SKOV3细胞的凋亡率分别为3.4%、20.1%;卡铂+PDTC作用24h、48h后卵巢癌SKOV3细胞的凋亡率明显增加,分别为14.0%、32.7%,与卡铂组相比有显著性差异(P<0.01)。卡铂作用3d、5d后卵巢癌SKOV3细胞中survivinmRNA的相对表达量分别为0.48±0.04、0.57±0.03,卡铂+PDTC组分别为0.33±0.04、0.38±0.04,对应时间点两组差异显著(P<0.01)。[结论]卡铂能诱导NF-κB的活化,NF-κB抑制剂可以增强卡铂诱导的SKOV3细胞的凋亡。NF-κB可能通过上调SKOV3细胞内survivinmRNA的表达来抵抗卡铂诱导的SKOV3细胞的凋亡。
[Objective] To investigate the toxic effect of pyrrolidine dithiocarbamate (PDTC) -treated carboplatin on ovarian cancer cells and its mechanism. [Methods] EMSA was used to observe the activation of NF-κB under the action of carboplatin and carboplatin and PDTC. MTT and flow cytometry were used to compare the expression of SKOV3 cells Growth and apoptosis rate. The expression of survivin mRNA was detected by RT-PCR. [Results] The results of EMSA showed that carboplatin induced NF-κB activation and PDTC could inhibit this effect. PDTC pretreatment can enhance the inhibitory effect of carboplatin on ovarian cancer SKOV3 cells. The apoptotic rates of ovarian cancer SKOV3 cells were respectively 3.4% and 20.1% at 24h and 48h after carboplatin treatment. The apoptotic rate of ovarian cancer SKOV3 cells was significantly increased after carboplatin and PDTC treatment for 24h and 48h (14.0% and 32.7%, respectively) , Significant difference compared with carboplatin group (P <0.01). The relative expression levels of survivin mRNA in ovarian cancer cell line SKOV3 after 3 and 5 days of carboplatin were 0.48 ± 0.04 and 0.57 ± 0.03, respectively, while those in carboplatin and PDTC group were 0.33 ± 0.04 and 0.38 ± 0.04 respectively P <0.01). [Conclusion] Carboplatin can induce the activation of NF-κB, and the inhibitor of NF-κB can enhance the apoptosis of SKOV3 cells induced by carboplatin. NF-κB may resist the apoptosis of SKOV3 cells induced by carboplatin by up-regulating the expression of survivin mRNA in SKOV3 cells.