目的 探讨呫吨酮并吡啶衍生物5，9-二(2-吡咯烷基乙酰氨基)-7H-吡啶并呫吨-7-酮 (XP-16) 抗人胃癌MGC-803细胞作用及其可能作用机制。方法 通过MTT法、细胞形态学和克隆实验观察XP-16对MGC-803细胞增殖;应用Hoechest33258和PI双染法观察细胞凋亡;采用荧光分光光度计检测细胞内钙(i)及线粒体膜电位;RT-PCR检测Bad和金属硫蛋白1A (MT-1A) mRNA的表达。结果 XP-16能抑制MGC-803细胞增殖，呈浓度(r=0.88， P<0.01)和时间(r=0.93， P<0.05)依赖性。XP-16作用MGC-803细胞24 h后，MGC-803细胞出现染色质聚集、核碎裂等典型的凋亡形态学改变，且随XP-16浓度的增加，MGC-803细胞凋亡百分率逐渐增大。XP-16作用后，MGC-803细胞的i和线粒体膜电位降低、Bad和MT-1A mRNA表达增加。结论 XP-16可通过诱导细胞凋亡抑制MGC-803细胞增殖，其机制可能与其降低i和线粒体膜电位有关，而细胞内MT-1A表达的上调可能是i下降的结果。
Abstract
OBJECTIVE To investigate aticancer effect and potential mechanism of a new xanthono-pyridine derivative N,N′-(7-oxo-7H-chromenoquinoline-5,9-diyl)-bis(2-(pyrrolidin-1-yl)acetamide) (XP-16) on human gastric carcinoma cell line MGC-803. METHODS Antiproliferative effect of XP-16 on MGC-803 cells was evaluated by MTT assay, morphological examination and colonial assay. Apoptosis detection was carried out using Hoechest33258 and PI double-dyeing method. Intracellular calcium concentration (i) and mitochondria membrane potential were detected by fluorospectrophotometer. Bad and metallothionein 1A (MT-1A) transcripts were analyzed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS XP-16 could inhibit proliferation of MGC-803 cells in dose- and time-dependent manner(r=0.88, P<0.01; r=0.93, P<0.05). Typical apoptotic morphology such as chromatin aggregation and nuclear fragmentation was observed in MGC-803 cells with XP-16-treated for 24 h, and the effect was dose-dependent. After treated with XP-16, i and mitochondria membrane potential of MGC-803 cells were decreased, relative mRNA levels of Bad and MT-1A were up-regulated. CONCLUSION XP-16 can inhibite MGC-803 cells proliferation by inducing apoptosis, which might be associated with decreasing i and mitochondria membrane potential. Up-regulation of MT-1A expression might be the result of decreased i.
关键词
5 /
9-二(2-吡咯烷基乙酰氨基)-7H-吡啶并呫吨-7-酮 /
人胃癌MGC-803细胞 /
凋亡 /
细胞内钙 /
线粒体膜电位 /
金属硫蛋白1A
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Key words
N,N′-(7-oxo-7H-chromenoquinoline-5,9-diyl)-bis(2-(pyrrolidin-1-yl)acetamide) /
human gastric carcinoma cell line MGC-803 /
apoptosis /
intracellular calcium /
mitochondria membrane potential /
metallothionein 1A
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中图分类号：
R965
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参考文献
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脚注
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基金
国家自然科学基金资助项目(21002015)；广西自然科学基金资助项目(2010GXNSFB013013, 0639030)
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