诱导型一氧化氮合酶对乳腺癌细胞化学敏感性的影响  

Effect of inducible nitric oxide synthase (iNOS) on chemosensitivity of a breast cancer cell MDA-MB-231^(wt)

作　　者：孔肇路[1] 金一尊[1] 

机构地区：[1]复旦大学放射医学研究所,上海200032

出　　处：《复旦学报（医学版）》2006年第6期732-735,765,共5页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金项目(30500143);国家863计划(2002AA2Z3104)资助

摘　　要：目的分析可诱导一氧化氮合酶(iNOS)对乳腺癌细胞化学敏感性的影响,评价新型蒽醌类生物还原化合物RH1对乳腺癌细胞毒性和乏氧选择性。方法实验以人乳腺癌细胞MDA-MB-231wt和其转染空载体的对照组细胞(PEF-vector)及一氧化氮合酶基因转染的细胞克隆(i NOS10)为实验对象。噻唑蓝(MTT)法比较不同酶活性的细胞克隆对顺铂等抗肿瘤药物和丝裂霉素C(MMC)衍生物-2,5-二氮丙啶-6-羟甲基-3-甲基苯-1,4-二酮(RH1)的敏感性的差异。结果同对照组细胞相比,i NOS10细胞对顺铂等几种抗肿瘤药物呈现不同程度的耐受,而对阿霉素的敏感性有所增强(P<0.05),用L-甲基盐酸精氨酸抑制细胞中NO的合成,则会降低i N-OS10细胞对阿霉素的敏感性,而对其他几种抗肿瘤药物的敏感性无显著影响;生物还原化合物RH1是高毒性的化合物,IC50较MMC下降577倍;乏氧条件下,细胞对RH1的化学敏感性显著增强,流式细胞术显示RH1可以引起i NOS10细胞周期的G2/M阻滞。结论MMC衍生物RH1是高细胞毒性的化合物,可诱导一氧化氮合酶活性的增强,也可引起肿瘤细胞对部分抗肿瘤药物的敏感性降低,且对高表达i NOS的细胞具有乏氧选择性的损伤作用。Purpose To examine the effect of iNOS on tumour cells chemosensitivity to a series of antineoplastic drugs in vitro and to clarify the possible role of iNOS in the metabolism of 2,5-biaziridi- nyl-6-hydroxy-methyl-3-methylbenzene-1,4-dione （RH1）, a new synthesized bioreductive compound. Methods Human breast cancer cell line （MDA-MB-231^wt） and its iNOS gene transfected clones （iNOS10） were exposed to aforementioned anticancer drugs and RH1. 3-[-4,5 dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide （MTT） assay were used to determine cells sensitivity, the change of cell cycle different timepoint after exposed in 0.01μmol/L RH1 were plot by flow cytometric analysis. Results iNOS-gene transferred cells（iNOS10） showed resistance to CDDP, MMC, mephalan and mitoxantrone compared to parent cells MDA-MB-231^wt and control cells PEF. However, iNOS10 cells were more sensitive to ADM. On the other hand, the toxicity of RH1 was more severe than MMC. RH1 enhanced cytotoxicity under hypoxia and could cause cell block in G2/M phase 24 hours after it was administrated. Conclusions Cells with high iNOS activity showed resistance to part of the antineoplastic drugs, it might because that iNOS could catalyse drugs into their non- or low-toxic production. And in the metabolism activation of RH1, iNOS could enhance or decrease the cytotoxicity of RH1 depending on the intracellur PO2, which caused higher cytotoxicity of RH1 to hypoxia cells with the increasing of iNOS activity.

关 键 词：生物还原 一氧化氮合酶 化学敏感性 抗肿瘤药物 

分 类 号：R730.53[医药卫生—肿瘤] R965.1[医药卫生—临床医学]