呫吨酮并吡啶衍生物XP-16诱导人肺癌A549细胞凋亡

XP-16,a new xanthono-pyridine derivative,induces apoptosis in human lung carcinoma A549 cells

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[4,3-c]呫吨-7-酮(XP-16)对人肺癌A549细胞的抗肿瘤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A549细胞增殖的影响;应用Hoechst 33258和PI双染法观察细胞凋亡;荧光分光光度计检测细胞内钙([Ca2+]i)及线粒体膜电位;qRT-PCR检测Bad和金属硫蛋白1A(metallothionein 1A,MT-1A)mRNA表达。结果 XP-16能抑制A549细胞增殖,呈剂量和时间依赖性。XP-16作用A549细胞24 h后,A549细胞出现染色质聚集、核碎裂等典型的凋亡形态学改变;随XP-16剂量的增加,A549细胞凋亡百分率逐渐增大。XP-16作用后,A549细胞的[Ca2+]i和线粒体膜电位降低、Bad和MT-1A mRNA的表达增加。结论 XP-16具有诱导A549细胞凋亡的作用,可能与其降低[Ca2+]i和线粒体膜电位有关。MT-1A表达的上调可能是[Ca2+]i降低的结果。

Aim To investigate the anticancer effect of a new xanthono-pyridine derivative N, N ＇-（ 7-oxo-7H-chromeno[3,2-h] quinoline-5,9-diyl）-bis（2-（ pyrroli-din-1-yl）acetamide） （XP-16） on human lung carcino-ma cell line A549 and the potential mechanism. Meth-ods Antiproliferative effect of XP-16 on A549 cells was evaluated by MTT assay, morphological examina-tion and colonial assay. Apoptosis detection was car-ried out using Hoechst 33258 and PI double-dyeing method. Intracellular Ca2＋ concentration （ [ Ca2＋] i ） and mitochondria membrane potential were detected by fluorospectrophotometer. A549 cells treated with XP-16 were collected for Bad and metallothionein 1 A （ MT-1 A ） transcript analysis by real-time reverse tran-scriptase-polymerase chain reaction （ qRT-PCR） . Re-sults XP-16 inhibited A549 cell proliferation in dose-and time-dependent manner. Typical apoptotic morphology such as chromatin aggregation and nuclear fragmentation was observed in A549 cells treated with XP-16 for 24 h, and the apoptosis was showed in a dose-dependent manner. After treated with XP-16, [ Ca2＋] i and mitochondria membrane potential of A549 cells were decreased, and relative mRNA level of Bad and MT-1A was up-regulated. Conclusions XP-16 has anticancer effect on A549 cells through apoptosis, which might be associated with decreasing intracellular Ca2＋ concentration and mitochondria membrane poten-tial. Up-regulation of MT-1A expression might be the result of decreased [ Ca2＋] i .