Induction of apoptosis by anesthetics in the human cancer cell lines.

麻醉剂诱导人类癌细胞系凋亡。

• 批准号: 13671609
• 负责人: NAGASAKA Hiroshi
• 金额: $ 1.28万
• 依托单位: Meikai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671609/
• 关键词: Apoptosis; Morphione; Codeinone; Cancer; アポトシス; コデイン

There are relatively few studies about antiproliferative effects of codeinone-related compounds on human cancer cell lines, compared with those of morphine related compounds. The authors previously found that codeinone, an oxidation metabolite of codeine, among 10 opiois, showed the highest cytotoxic activity (DNA fragmentation-inducing activity) agains human promyelocytic leukemic cell lines (HL-60). This was counteracted by an antioxidant, N-acetyl-L-cysteine (NAC). These finings prompted us performed a more detailed study of apoptosis induction after codeinone treatment. Apoptosis was induced by treating HL60 cells for 1-6h with codeine or codeinone. DNA after staining with diamidinopheylindole (DAPI). The appearance of apoptotic cells was monitored bymicroscopic observation after staining with Hoechst (H)-33342,and fluorescence actiated cell sorter (FACS) after staining with Annexin. The release of cytochrome C and cytocrome oxidase from mitochondria and activation of caspase 3 were monitored by Western blot analysis. Intracellular caspase 3 like activity was confirmed by FACS, using cell permeable substate. Mitochondrial manganese-containing superoxide dismutase (MnSOD) activity and mRNAexpression were assayed by ativity staining after separation on the polyacrylamide gel electrophoresis, and reverse transcriptase-poly-merase chain reaction (RT-PCR), respectively. Codeinone induced internucleosomal DNA fragmentation and production of Annexin-positive apoptotic cells more potently than codeine in HL-60 cells. Codeinone stimulated the release of both cytochrome C and cytochrome oxidase, and cleavage of procaspase 3 without significant changes in both the activity and expression of MnSOD. Codeinone was found to possess both apoptosis and necrosis-inducing activity, in addition to the reported antinociceptive activity, further substantiated its antitumor potential.

与吗啡相关化合物相比，可待因酮相关化合物对人癌细胞系的抗增殖作用的研究相对较少。作者先前发现，在10种阿片类药物中，可待因的氧化代谢产物可待因酮对人早幼粒细胞白血病细胞系（HL-60）显示出最高的细胞毒活性（DNA断裂诱导活性）。这是抵消了抗氧化剂，N-乙酰-L-半胱氨酸（NAC）。这些发现促使我们对可待因酮处理后的细胞凋亡诱导进行了更详细的研究。用可待因或可待因酮处理HL 60细胞1- 6 h，诱导细胞凋亡。用二脒基苯并吲哚（DAPI）染色后的DNA。Hoechst（H）-33342染色后通过显微镜观察，Annexin染色后通过荧光激活细胞分选仪（FACS）监测凋亡细胞的出现。Western印迹分析检测线粒体细胞色素C和细胞色素氧化酶的释放以及caspase 3的活化。流式细胞仪检测细胞内caspase 3样活性。采用聚丙烯酰胺凝胶电泳分离后活性染色法和逆转录-聚合酶链反应（RT-PCR）法分别检测线粒体锰超氧化物歧化酶（MnSOD）活性和mRNA表达。可待因酮诱导HL-60细胞核小体间DNA断裂和Annexin阳性凋亡细胞的产生比可待因更强。可待因酮刺激细胞色素C和细胞色素氧化酶的释放，并在MnSOD的活性和表达没有显着变化的半胱氨酸天冬氨酸蛋白酶3的裂解。可待因酮除了具有抗伤害活性外，还具有诱导细胞凋亡和坏死的活性，进一步证实了其抗肿瘤的潜力。

项目成果

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