CAJ | 학술논문

目的研究拓扑异构酶Ⅰ抑制剂对耐阿霉素淋巴瘤细胞株(L1210/ADM)的杀伤和诱导凋亡的作用.方法采用噻唑蓝(MTT法)、钙依赖性磷脂结合蛋白(Annexin V)染色、梯状DNA电泳与细胞形态学观察等方法分析不同浓度(0.05、0.10、0.20、0.40μmol/L)拓扑异构酶Ⅰ抑制剂对L1210/ADM细胞的杀伤和凋亡作用,采用半胱氨酸天冬氨酸蛋白酶(Caspase)8、Caspase-3抑制剂IETD-fmk、DEVD-CHO分析拓扑异构酶Ⅰ抑制剂介导的靶细胞杀伤和凋亡作用与Caspase的关系.结果 0.10 μmol/L拓扑异构酶Ⅰ抑制剂对靶细胞株的杀伤作用,与对照组和0.05 μmol/L组之间差异均有统计学意义(均P＜0.05).加用Caspase酶特异性抑制剂3组各时间点的靶细胞存活率均高于单用拓扑异构酶Ⅰ抑制剂组,差异均有统计学意义(均P＜0.05).靶细胞株经拓扑异构酶Ⅰ抑制剂处理后,部分靶细胞株出现了细胞凋亡;加用Caspase酶特异性抑制剂组中,有部分细胞死亡,但无凋亡小体形成,死亡细胞均为锥虫蓝染色.经拓扑异构酶Ⅰ抑制剂处理的靶细胞株大部分细胞出现胞体解离,并彼此黏附成簇状的、锥虫蓝拒染的凋亡小体,靶细胞株DNA电泳表现为特征性梯状图谱;加入Caspase酶特异性抑制剂的各组可以看到阻断了梯状DNA的产生.结论拓扑异构酶Ⅰ抑制剂对L1210/ADM细胞株具有较强的杀伤和诱导凋亡作用,其机制可能涉及Caspase-8、Caspase-3的有序性活化.
목적 연구탁복이구매Ⅰ억제제대내아매소림파류세포주(L1210/ADM)적살상화유도조망적작용.방법 채용새서람(MTT법)、개의뢰성린지결합단백(Annexin V)염색、제상DNA전영여세포형태학관찰등방법분석불동농도(0.05、0.10、0.20、0.40μmol/L)탁복이구매Ⅰ억제제대L1210/ADM세포적살상화조망작용,채용반광안산천동안산단백매(Caspase)8、Caspase-3억제제IETD-fmk、DEVD-CHO분석탁복이구매Ⅰ억제제개도적파세포살상화조망작용여Caspase적관계.결과 0.10 μmol/L탁복이구매Ⅰ억제제대파세포주적살상작용,여대조조화0.05 μmol/L조지간차이균유통계학의의(균P＜0.05).가용Caspase매특이성억제제3조각시간점적파세포존활솔균고우단용탁복이구매Ⅰ억제제조,차이균유통계학의의(균P＜0.05).파세포주경탁복이구매Ⅰ억제제처리후,부분파세포주출현료세포조망;가용Caspase매특이성억제제조중,유부분세포사망,단무조망소체형성,사망세포균위추충람염색.경탁복이구매Ⅰ억제제처리적파세포주대부분세포출현포체해리,병피차점부성족상적、추충람거염적조망소체,파세포주DNA전영표현위특정성제상도보;가입Caspase매특이성억제제적각조가이간도조단료제상DNA적산생.결론 탁복이구매Ⅰ억제제대L1210/ADM세포주구유교강적살상화유도조망작용,기궤제가능섭급Caspase-8、Caspase-3적유서성활화.
Objective To observe the killing and proapoptotic effects of topoisomerase Ⅰ (topotecan) inhibitor on L1210/ADM cells. Methods To analyze the the killing and proapoptotic effects of topoisomerase Ⅰ on L1210/ADM cells with MTT method, Annexin V, DNA ladder electrophoresis and cell morphology analysis. The relation between the killing and proapoptotic effects and Caspase using Caspase-8, inhibitor IETD-fmk and DEVDCHOs was observed. Results In contrast with 0. 05 μ mol/L group, the killing and proapoptotic effects of 0.10 μmol/L topoisomerase inhibitor was statistically different. With 3 groups of Caspase enzyme inhibitor at each time point, survival rate of target topoisomerase group was higher than that of the single-use topoisomerase Ⅰ inhibitor group(P ＜0.05). After treatment with inhibitors of topoisomerase Ⅰ , some target cells died. In Caspase enzyme inhibitor group, some cells died but no apoptotic body formation, dead cells were trypan blue stained. Majority of target cells ever incubated by the topoisomerase Ⅰ inhibitors got dissociated and tufted with each other. Apoptotic bodies formed without trypan blue staining. Target cells electrophoresis showed the characteristic of DNA ladder pattern in the group with Caspase specific inhibitor of the enzyme can block the formation of DNA ladder. Conclusion Topoisomerase Ⅰ inhibitor has significant killing and proapoptotic effects on L1210/ADM cell lines, which can depend on activation of Caspase-8 and Caspase-3.