尿多酸肽（CDA2）诱导多发性骨髓瘤细胞RPMI8226凋亡实验探究.docVIP

尿多酸肽（CDA2）诱导多发性骨髓瘤细胞RPMI8226凋亡实验探究 　 作者：叶宝东 沈一平 周郁鸿 邵科钉 俞庆宏 【摘要】 　　［目的］观察尿多酸肽（CDA2）对多发性骨髓瘤细胞RPMI8226抑制增殖和凋亡作用。［方法］应用MTT法，检测CDA2对多发性骨髓瘤细胞株RPMI8226增殖的影响。通过Hochest33258染色、流式细胞仪（Annexin V和PI双染色）检测细胞凋亡。［结果］CDA2明显抑制RPM18226细胞的增殖。CDA2作用RPM18226细胞增殖24h的半数抑制浓度（IC50）分别为1.64mg/g。采用Hochest33258染色，荧光显微镜下观察CDA2以2mg/g及4mg/g终浓度作用于RPM18226 24h后细胞形态改变，可见明显的凋亡小体出现。Annexin VPI双标记法流式细胞仪分析结果显示，RPMI8226细胞2mg/gCDA2作用6、12、24h后，早期凋亡细胞百分数分别为5.43%、12.02%、29.07%，明显高于空白对照组4.88%。［结论］CDA2对MM细胞株RPMI8226有生长抑制作用，可能是通过诱导其发生凋亡而起作用。 【关键词】 多发性骨髓瘤；尿多酸肽；细胞凋亡 　　Abstract：［Objective］To investigate the proliferation inhibition and apoptosisinducing effect of uroacitides（CDA2）on human multiple myeloma RPMI8226 cells.［Methods］MTT assay was undertaken to observe the growth inhibition of RPMI8226 cells exposed to CDA2.The apoptosis of RPMI8226 cells were detected by fluorescence staining with hochest33258，flow cytometry（Annexin V and PI staining）.［Results］MTT analysis indicated that the proliferation of RPMI8226 cells were inhibited significantly with CDA2 treatment for 24h.Treatment with different concentrations of CDA2 for 24h，50% growth inhibition（IC50）at 24h in RPMI8226 was 1.64mg/g.Cell appearances were observed by fluorescence staining with hochest33258 after CDA2（2mg/g and 4mg/g）for RPMI8226，we could see apoptotic body evidently with fluorescence microscope.After treatment with concentrations of CDA2（2mg/g）for 6h，12h，24h，RPMI8226 had 5.43%，12.02% and 29.07% apoptotic cells，which was more than that of untreated group（4.88%）.［Conclusions］Uroacitides（CDA2）could inhibit the proliferation and induce apoptosis of human multiple myeloma RPMI8226 cells in vitro. 　　Key words：multiple myeloma；CDA2；cell apoptosis 尿多酸肽（CDA2）是从健康人尿中提取的含有多种有机酸和多肽成分的非细胞毒性新型诱导分化治疗抗癌药［1］。近年来的研究表明，CDA2对多种肿瘤细胞具有诱导分化和凋亡作用，而且还可以逆转肿瘤耐药，防止肿瘤复发和转移［2］。但对多发性骨髓瘤（Multiple Myeloma，MM）的作用尚未见报道，我们观察了CDA2对多发性骨髓瘤细胞RPMI8226增殖抑制的作用，并初步从诱导凋亡角度探讨其作用机制，为临床应用CDA2治疗提供多发性骨髓瘤实验依据。 　　1 材料和方法 　　1.1 细胞株 　　人多发性骨髓瘤细胞株：RPM18226（浙江大学第一附属医院血液研究所惠赠）。