Induction of apoptosis by anesthetics I n the human cancer cell lines

研究人员对吗啡在临床浓度（10^-8 M）下诱导细胞凋亡的能力进行了研究。细胞毒性用3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四唑溴化盐（MTT）检测，用Annexin V和碘化丙啶（PI）通过荧光激活细胞分选仪（FACS）分析诱导早期凋亡和坏死，通过特异性底物的切割激活caspase-2、-3、-8和-9，琼脂糖凝胶电泳DNA片段化，自由基强度和O_2清除活性。与正常人细胞相比，毫摩尔浓度的吗啡对人肿瘤细胞系（HL-60，A549，MCF 7）的细胞毒性更高。临床浓度的吗啡在HL-60和A549细胞中产生早期凋亡标志物，而在MCF 7细胞中诱导更多的坏死细胞。临床浓度的吗啡不能激活任何半胱天冬酶，仅诱导微量的核小体间DNA断裂。 关于我们 与吗啡的细胞毒性浓度相反，吗啡镇痛研究可能为癌症的治疗和预防提供新的策略，使用临床浓度的吗啡不仅作为抗伤害感受剂，而且作为凋亡或坏死诱导剂。此外，可待因的氧化代谢产物可待因酮的可能的诱导活性，没有或与抗癌药物，进行了研究。可待因酮在HL-60中诱导核小体间DNA断裂，但在HSC-2中不诱导。可待因酮剂量依赖性地激活这两种细胞中的caspase-3，但程度远低于放线菌素D。可待因酮的这种性质与我们以前在α，α，β-不饱和酮中发现的类似。可待因酮在这些细胞中不激活caspase-8或caspase-9。N-乙酰-L-半胱氨酸抑制可待因酮对HSC-2细胞的细胞毒活性，抗坏血酸钠等则有一定的相加作用。此外，还研究了吗啡的氧化代谢产物吗啡酮在HL-60细胞中是否也引起类似的细胞死亡。因此，我们的数据表明，吗啡酮诱导HL-60细胞的非凋亡性细胞死亡。少

An investigation of the ability of morphine to induce apoptosis at its clinical concentration (10^-8M) was undertaken. Cytotoxicity was tested by3-[4, 5-dimethylthiazol-2-y1]-2, 5-diphenyltetrazolium bromide (MTT), assay, induction of early apoptosis and necrosis by fluoresxence-activated cell sorter (FACS) analysis with Annexin V and propidium iodine (PI), activation of caspase-2, -3, -8 and -9 by cleavage of specific substrates, DNA fragmentation by agarose gel electrophoresis, radical intensity and 0_2 scavenging activity by ESR spectroscopy. Millimolar concentrations of morphine showed higher cytotoxicity against human tumor cell lines (HL-60, A549, MCF7) than against normal human cells. The clinical concentration of morphine produced early apoptotic markers in HL-60 and A549 cells whereas it induced higher numbers of necrotic cells in MCF 7 cells. The clinical concentration of morphine failed to activate any caspase species and induced only trace amounts of internucleosomal DNA fr … More agmentation, in contrast to cytotoxic concentration of morphine. Abovementioned study may offer new strategies for treatment and prevention of cancer using a clinical concentration of morphine not only as an anti-nociceptive, but also as an apoptosis or necrosis inducer. In addition, the possible-inducing activity of codeinone, an oxidative metabolite of codeine, without or with anticancer drugs, was investigated. Codeinone induced internucleosomal DNA fragmentation in HL-60, but not in HSC-2. Codeinone dose-dependently activated caspase-3 in both of these cells, but to a much lesser extent than attained by actinomycine D. This property of codeinone was similar to what we have found previously in a, α, β-unsaturated ketones. Codeinone did not activate caspase-8 or caspase-9 in these cells. The cytotoxic activity of codeinone against HSC-2 cells was inhibited by N-acetyl-L-cysteine, but somewhat additively stimulated by sodium ascorbate, etc. Moreover, Whether, morphinone, an oxidative metabolite of morphine, also induced a similar type of cell death in HL-60 cells was investigated. As results, our data suggest that morphinone induces non-apoptotic cell death in HL-60 cells. Less