CAJ | 학술논문

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。
목적：탐토첩둔동병필정연생물5,9-이(2-필각완기을선안기)-7H-필정병[4,3-c]첩둔-7-동(XP-16)대인폐암A549세포적항종류작용급기가능작용궤제。방법통과MTT법、세포형태학화극륭실험관찰XP-16대A549세포증식적영향；응용Hoechst 33258화PI쌍염법관찰세포조망；형광분광광도계검측세포내개([Ca2+]i)급선립체막전위；qRT-PCR검측Bad화금속류단백1A(metallothionein 1A,MT-1A) mRNA표체。결과 XP-16능억제A549세포증식,정제량화시간의뢰성。 XP-16작용A549세포24 h후,A549세포출현염색질취집、핵쇄렬등전형적조망형태학개변；수XP-16제량적증가,A549세포조망백분솔축점증대。 XP-16작용후,A549세포적[ Ca2+] i 화선립체막전위강저、Bad 화MT-1A mRNA적표체증가。결론 XP-16구유유도A549세포조망적작용,가능여기강저[ Ca2+] i 화선립체막전위유관。 MT-1A표체적상조가능시[Ca2+]i 강저적결과。
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 mor- <br> phology 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 .