探讨重组改构人肿瘤坏死因子（recombinant mutant human tumor necrosis factor, rmh TNF）体外逆转人卵巢癌多药耐药细胞株SKOV3/DDP的耐顺铂（cisplatin,又称DDP）效应及其可能存在的作用机制。方法: 体外培养人卵巢癌多药耐药细胞株SKOV3/DDP，以MTT法检测rmh TNF对此耐药株的细胞毒作用，确定其对此细胞株的无毒剂量;同法测定无毒剂量rmh TNF干预后SKOV3/DDP对顺铂耐药性的变化，通过流式细胞术检测rmh TNF干预不同时间段SKOV3/DDP细胞株中GST π蛋白的表达，RT PCR方法分析rmh TNF处理SKOV3/DDP细胞前后 MDR1基因的表达水平。结果: （1）50～12234 U/ml rmh TNF对SKOV3/DDP细胞增殖无明显抑制作用（细胞存活率均﹥90%），因此选用100 U/ml 作为逆转剂量（无毒剂量）;（2）单用DDP 24、48、72 h 的IC 50 为(23.29±0.43)、(8.97±0.69)、(6.43±0.79)μg/ml，联合100 U/ml rmh TNF和DDP用药使SKOV3/DDP的IC50分别降至（19.50±0.50）、（4.34±0.43）、（2.44±0.02）μg/ml，逆转倍数分别为1.19、206、2.64倍;（3）100 U/ml rmh TNF干预0、24、48、72 h后SKOV3/DDP细胞内GST π蛋白的表达随作用时间延长降低， MDR1基因的表达随作用时间延长降低。结论: rmh TNF对SKOV3/DDP有逆转耐药作用，其机制可能与下调GST π蛋白及 MDR1基因表达有关。

To investigate the reversing effect of recombinant mutant human tumor necrosis factor (rmh TNF) on cisplatin(DDP) resistant human ovarian cancer cell line SKOV3/DDP in vitro and the related mechanism. Methods: DDP resistant human ovarian cancer cell line SKOV3/DDP was cultured in vitro. The cytotoxic effect of rmh TNF to SKOV3/DDP cells was examined by MTT assay and the nontoxic dose of rmh TNF was identified. The changes of DDP resistance was observed after cells were treated with nontoxic dose of rmh TNF by MTT assay. The expre ssion of GST π protein was examined by flow cytometry at different periods after rmh TNF intervention; RT PCR was used to analyze the expression of MDR1 gene in SKOV3/DDP cells before and after rmh TNF treatment. Results:（1）rmh TNF at 50 122.34 U/ml showed no evident inhibitory effect on the growth of SKOV3/DDP cells (the cell survival rate higher than 90%); and 100 U/ml was chosen for the reversing experiment ( nontoxic dose).（2）IC 50 values of SKOV3/DDP cells were (2329±0.43), (8.97±0.69) and (6.43±0.79) μg/ml after treatment with DDP for 24, 48 and 72 h, respectively; and the values decreased to （19.50±0.50）,（4.34±0.43） and （2.44±0.02）μg/ml after combined treatment with 100 U/ml rmh TNF, respectively.（3）Expression of GST π protein and MDR1 gene decreased with the prolongation of rmh TNF treatment. Conclusion: rmh TNF has reversal effect on the DDP resistant cell line SKOV3/DDP, and the mechanism may be associated with the down regulation of GST π protein and MDR1 gene expression.

河北省科技攻关计划（No.072761147）