pkf118310对k562细胞增殖和凋亡的影响及其机制研究-effects of pkf 118310 on proliferation and apoptosis of k562 cells and its mechanism.docxVIP
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pkf118310对k562细胞增殖和凋亡的影响及其机制研究-effects of pkf 118310 on proliferation and apoptosis of k562 cells and its mechanism
目录中文摘要 1Abstract2缩略词表3前言 4PKF118-310 对 K562 细胞增殖和凋亡的影响及其机制研究14研究对象和方法 14结果 18讨论 22结论 24参考文献25综述28攻读学位期间的研究成果33 致谢34学位论文原创性声明35PKF118-310对K562细胞增殖和凋亡的影响及其机制研究导师:杜新教授摘要【目的】探讨PKF118-310对人慢性粒细胞白血病( CML ) K562细胞增殖及凋亡的影响及 其机制研究。【方法】用不同浓度PKF118-310处理K562细胞,MTT法检测药物对细胞的抑制作用;免 疫荧光方法观察细胞核中β-catenin/TCF转录复合物的形成;流式细胞术检测细胞凋亡; Western印迹法检测caspase3、caspase8、XIAP、Bcl-2、Bcl-9、β-catenin、TCF-4、c-myc 及cyclin D1的表达。【结果】 PKF118-310能够抑制K562细胞生长,对K562细胞在24小时的IC50为5.388umol/L,48小时的IC50为3.290umol /L,72小时的IC50为1.566umol /L。免疫荧光证实细胞核 内形成β-catenin/TCF转录复合物。分别用1.6及3.2umol/l的PKF118-310作用于K562细胞 48小时后检测细胞凋亡,Annexin V-FITC /PI 双标法显示其凋亡率分别为( 48.0± 0.9) %、( 80.2± 1. 2) %、均高于对照组( 1.2± 0. 6 ) % ( P < 0. 05) 。PKF118-310处理K562细胞72h后,caspase3及caspase8蛋白表达升高,而XIAP,Bcl-2、Bcl-9、β-catenin、 TCF、c-myc及cyclin D1蛋白表达明显降低。【结论】PKF118-310 可抑制 K562 细胞增殖及诱导细胞凋亡。【关键词】CML;Wnt/β-catenin信号通路;PKF118-310- 1 -Effects and its mechanism of PKF118-310 onproliferation and apoptosis of K562 cellsABSTRACT[Objective] To investigate the effects and its mechanism of PKF118-310 on proliferation and apoptosis of chronic myeloid leukemia cell line K562.[Methods] After treatment of PKF118-310 at the different concentration, the proliferation inhibition on K562 cells was detected by MTT, whether the β-catenin/TCF transcription complex was existed in the nucleus was observed by immunofluorescence. Cell apoptosiswas detected by flow cytometry. The expressions of caspase3,caspase8,XIAP,Bcl-2,Bcl-9,β-catenin,TCF,c-myc and cyclin D1 were detected by Western blot.[Results] PKF118-310 can inhibit the proliferation of K562 cells. The median inhibitory concentration (IC50) of PKF118-310-treated K562 cells for 24h, 48h, and 72h was 5.388umol/L, 3.290umol/L, 1.566umol/L respectively. The β-catenin/TCF transcription complex inthe nucleus was observed by immunofluorescence. The results showed that after being treated with 1.6 and 3.2umol/l PKF118-310 for 48h,the inhibitory rates of K562 cells were ( 48.0±0. 9) %,( 80.2± 1. 2) %,respectiv
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