NF-κB抑制剂二硫代氨基甲酸吡咯烷提高卵巢癌细胞顺铂化疗的敏感性

Pyrrolidine dithiocarbamate,a inhibitor of NF-κB,enhances chemosensitivity of ovarian cancer cells to cisplatin

目的:探讨核因子κB(nuclear factor kappa B,NF-κB)抑制剂二硫代氨基甲酸吡咯烷(pyrrolidine dithiocarbamate,PDTC)提高人卵巢癌SKOV-3细胞对顺铂(cisplatin)化疗敏感性的作用及其可能的机制。方法:用不同浓度PDTC联合顺铂在不同时间作用于卵巢癌SKOV-3细胞,MTT法观察药物作用对细胞生长的抑制,Western Blotting分析胞质内NF-κB抑制蛋白α(inhibitor of NF-κB,I-κBα)、胞核P65蛋白的表达;流式细胞术检测细胞凋亡。结果:PDTC(10 ～40μmol/L)或顺铂(0.1 ～100μg/ml)均明显抑制SKOV-3细胞的生长(P<0.05或P<0.01),并引起细胞的凋亡;小剂量PDTC(2.5、5μmol/L)和顺铂(0.01μg/ml)联合应用与单用顺铂比较,可明显增加细胞生长抑制率和细胞凋亡率(均P<0.05)。单用顺铂组与对照组比较,胞质I-κBα蛋白减少而胞核P65蛋白增多,联合使用PDTC可逆转此现象。结论:小剂量PDTC可增强卵巢癌细胞对顺铂的敏感性,这一作用可能与PDTC增加I-κBα蛋白的表达而抑制P65蛋白进入核内有关。

Objective： To determine the promoting effect of pyrrolidine dithiocarbamate （PDTC）, an inhibitor of NF- κB, on the sensitivity of human ovarian cancer cell line SKOV-3 to chemoagent cisplatin and the possible mechanisms. Methods： SKOV-3 cells were treated with various concentrations of cisplatin and PDTC combination for different periods. Then the growth inhibition of SKOV-3 cells were observed by MTT assay, expression of I-κBα and P65 protein by Western blotting assay, and apoptosis of cells by flow cytometry. Results： Separate application of both cisplatin（0.1-10 μg/ml） and PDTC （ 10 - 40μmol/L L） significantly inhibited the growth of SKOV-3 ceils and caused cell apoptosis （ P 〈 0.05 or P 〈 0.01 ）. Even at lower concentration, cisplatin （0.01 μg/ml） combined with PDTC （2.5, 5μmol/L） resulted more sig- nificant growth inhibition and apoptosis of SKOV-3 cells than cisplatin alone （both P 〈 0.05）. Cisplatin alone reduced cytoplasmic I-κBα protein in the cytoplasm and increased nuclear P65 protein compared with control group（ both P 〈 0. 05 ）, which could be reversed by addition of PDTC. Conclusion ： Low dose of PDTC can enhance the chemosensitivity of ovarian cancer ceils to cisplatin, which might be related to PDTC-induced I-κBα protein and the subsequent inhibition of nuclear translocation of P65 protein.