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

研究了临床浓度(10^~(-8)M)吗啡诱导细胞凋亡的能力。用3-[4，5-二甲基四氮唑-2-y1]-2，5-二苯基四氮唑溴化法检测细胞毒性，Annexin V和PI荧光激活细胞分类器(FACS)分析早期细胞凋亡和坏死，特定底物裂解caspase-2，-3，-8和-9，琼脂糖凝胶电泳法测定DNA断裂，ESR波谱测定自由基强度和0_2清除活性。毫摩尔浓度的吗啡对人肿瘤细胞株(HL-60、A549、MCF7)的杀伤作用明显高于对正常人细胞的杀伤作用。临床浓度的吗啡在HL-60和A549细胞中产生早期的凋亡标志物，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氨酸天冬氨酸氨基转移酶，只能诱导微量的核小体间dna来自…。更多的凝集，与细胞毒性浓度的吗啡相反。上述研究可能为临床使用浓度的吗啡治疗和预防癌症提供新的策略，不仅作为抗伤害性药物，而且作为细胞凋亡或坏死的诱导剂。此外，还研究了可待因的氧化代谢产物可待酮在单独或与抗癌药物合用时可能的诱导活性。Codeinone诱导HL-60细胞核小体间DNA断裂，但不诱导HSC-2细胞DNA断裂。在这两种细胞中，可待酮都呈剂量依赖性地激活Caspase-3，但程度比放线菌D小得多。这种可待酮的性质类似于我们以前在α，β不饱和酮中发现的。Codeinone不能激活这些细胞中的caspase-8或caspase-9。N-乙酰-L-半胱氨酸抑制可待酮对HSC-2细胞的杀伤作用，抗坏血酸钠等对其有一定的刺激作用。此外，还研究了吗啡的氧化代谢产物吗啡酮是否也能诱导HL-60细胞死亡。结果表明，吗啡酮可诱导HL-60细胞发生非凋亡性死亡。较少

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