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生长抑制因子诱导膀胱肿瘤细胞凋亡的研究 被引量:1
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作者 李磊 贺大林 +4 位作者 何辉 王新阳 张林琳 罗勇 南勋义 《中华医学杂志》 CAS CSCD 北大核心 2005年第25期1766-1769,共4页
目的探讨生长抑制因子(PML)诱导膀胱肿瘤细胞凋亡的分子机制。方法应用脂质体Lipofectamine2000分别将重组可诱导表达的真核表达载体PMEP4/PML和对照PMEP4空载体转染到膀胱肿瘤细胞系UMUC2中,300μg/ml潮霉素B筛选稳定表达PML的抗性克... 目的探讨生长抑制因子(PML)诱导膀胱肿瘤细胞凋亡的分子机制。方法应用脂质体Lipofectamine2000分别将重组可诱导表达的真核表达载体PMEP4/PML和对照PMEP4空载体转染到膀胱肿瘤细胞系UMUC2中,300μg/ml潮霉素B筛选稳定表达PML的抗性克隆。激光共聚焦检测PML蛋白表达。DNAladder法检测肿瘤细胞凋亡。Western印迹法检测凋亡相关蛋白表达情况。结果经5μmol/L的CdSO4诱导表达后,激光共聚焦显微镜观察发现转染PMEP4/PML组的细胞核内有散在的斑点样的黄绿色荧光亮点,而转染空载体PMEP4组细胞未见特异性的荧光斑点表达。DNA梯度法显示转染PML细胞在诱导PML表达后24h出现梯状凋亡条带。Western印迹法检测凋亡相关蛋白半胱氨蛋白水解酶3、活化PARP上调,而survivin的表达水平下调。结论PML诱导膀胱肿瘤细胞凋亡可能与上调半胱氨蛋白水解酶3、活化PARP蛋白,抑制survivin的表达有关。 展开更多
关键词 肿瘤细胞凋亡 生长抑制因子 WESTERN印迹法 膀胱 半胱氨蛋白水解酶 survivin 凋亡相关蛋白 pml蛋白表达 激光共聚焦 真核表达载体 诱导表达 肿瘤细胞系 mol/L 分子机制 方法应用 抗性克隆 稳定表达 表达情况 细胞核内
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STUDY ON EFFECTS OF QUERCETIN ON PML GENE AND PROTEIN IN LEUKEMIA CELL LINES
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作者 钟璐 陈芳源 +2 位作者 欧阳仁荣 韩洁英 邵念贤 《Journal of Shanghai Second Medical University(Foreign Language Edition)》 2003年第1期12-15,29,共5页
Objective To investigate the effect of quercetin on PML gene and protein expression andlocalization in leukemia cell lines. Methods Cell morphology was assayed by Wright's stain and fluorescence stain, and PML mRN... Objective To investigate the effect of quercetin on PML gene and protein expression andlocalization in leukemia cell lines. Methods Cell morphology was assayed by Wright's stain and fluorescence stain, and PML mRNA expression by RT-PCR , PML protein localization by immuno-fluorescence. Results NB4 and HL-60 cells differentiated morphologically after treatment with all-trans-retinoic acid (ATRA) while K562 cells did not differentiate. Typical apoptosis was found in each cell line after treatment with quercetin. Immuno-fluorescence analysis showed , after treatment with ATRA , the fusion protein disappeared in NB4 cells and PML protein relocated , while HL-60 and K562 cells had no difference from control cells. After treatment with quercetin, the fusion protein disappeared in NB4 cells, PML protein relocated, then degraded. In HL-60 cells and K562 cells, PML protein also located and then degraded . The expression of PML mRNA was not changed in all three cell lines after treatment with ATRA or quercetin. Conclusion PML plays the role of differentiation and apoptosis inducer in leukemia cells at the translational level. PML in POD plays the role of apoptosis inducer and the growth control of leukemia cells. 展开更多
关键词 quercetin ATRA pml NB4 cell HL-60 cell K562 cell
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