重组改构人TNF逆转卵巢癌耐药细胞株SKOV3/DDP的耐药性及其机制  

Recombinant mutant human-TNF in reversing drug-resistance in ovarian cancer cell line SKOV3/DDP and the related mechanism

作　　者：魏素菊[1] 刘海英[1] 史健[1] 韩文峰[1] 

机构地区：[1]河北医科大学第四医院肿瘤内科,石家庄050051

出　　处：《中国肿瘤生物治疗杂志》2008年第2期150-154,共5页Chinese Journal of Cancer Biotherapy

基　　金：河北省科技攻关计划(No072761147)~~

摘　　要：目的：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）50～122.34U/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100U/ml作为逆转剂量（无毒剂量）;（2）单用DDP24、48、72h的IC50为（23.29±0.43）、（8.97±0.69）、（6.43±0.79）μg/ml,联合100U/ml rmh-TNF和DDP用药使SKOV3/DDP的IC50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）100U/ml rmh-TNF干预0、24、48、72h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。Objective：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 MDR1gene 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）IC50 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 MDR1gene 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 MDR1gene expression.

关 键 词：重组改构人肿瘤坏死因子 卵巢癌 多药耐药 逆转 GST-Π MDR1 

分 类 号：R737.31[医药卫生—肿瘤] R730.54[医药卫生—临床医学]