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

研究了吗啡在临床浓度（10^-8M）下诱导细胞凋亡的能力。采用3-[4,5 -二甲基噻唑-2-y1]- 2,5 -二苯基溴化四唑（MTT）检测细胞毒性，采用Annexin V和丙啶碘（PI）荧光活化细胞分选仪（FACS）检测细胞早期凋亡和坏死，通过切割特定底物激活caspase-2、-3、-8和-9，琼脂糖凝胶电泳激活DNA片段，ESR光谱检测自由基强度和0_2清除活性。毫摩尔浓度吗啡对人肿瘤细胞系（HL-60、A549、MCF7）的细胞毒性高于对正常人细胞的毒性。临床浓度吗啡在HL-60和A549细胞中产生早期凋亡标记，而在mcf7细胞中诱导更多的坏死细胞。与吗啡的细胞毒性浓度相比，临床浓度的吗啡未能激活任何半胱天蛋白酶，仅诱导微量的核体间DNA增强。上述研究可能为使用临床浓度的吗啡不仅作为抗伤害性药物，而且作为细胞凋亡或坏死诱导剂治疗和预防癌症提供新的策略。此外，我们还研究了可待因的氧化代谢物可待因酮在没有或有抗癌药物的情况下可能的诱导活性。可待因酮诱导HL-60核体间DNA断裂，HSC-2无。在这两种细胞中，可待因酮的剂量依赖性激活了caspase-3，但其激活程度远低于放线菌素d。可待因酮的这种特性与我们之前在a、α、β-不饱和酮中发现的相似。可待因酮不能激活这些细胞中的caspase-8或caspase-9。n -乙酰- l-半胱氨酸抑制了可待因酮对HSC-2细胞的细胞毒活性，而抗坏血酸钠等对其有一定的刺激作用。此外，我们还研究了吗啡的氧化代谢物吗啡酮是否也能诱导HL-60细胞类似类型的细胞死亡。结果表明，吗啡酮可诱导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