4-〔4″-(2″,2″,6″,6″-四甲基哌啶氮氧自由基)氨基〕-4′-去甲表鬼臼毒素诱导NB4肿瘤细胞凋亡

4-〔4″-(2″,2″,6″,6″-Tetramethyl-1″-piperidinyloxy)amino〕-4′-demethylepipodophyllotoxin inducing NB4 cell apoptosis

为了从细胞凋亡角度探讨 4-〔4″- ( 2″,2″,6″,6″-四甲基 - 1″-哌啶氮氧自由基 )氨基〕- 4′-去甲表鬼臼毒素 ( GP7)抗肿瘤作用的机理 ,用活细胞计数法及MTT比色法观察了 GP7对 NB4细胞的生长抑制作用 ;用光学显微镜及电子显微镜观察了凋亡细胞的形态学变化 ;用琼脂糖凝胶电泳观察了 DNA的梯形改变 .结果发现 :GP70 .1 8- 1 8μmol· L-1能显著抑制 NB4细胞的生长增殖 ;GP7可明显诱导NB4细胞凋亡的形态学变化及 DNA梯形带 .凋亡率随处理时间延长而增高 . 9μmol· L-1处理 NB4细胞 48h凋亡率达到最高峰 ,为 45.0± 3.0 ) % .延长处理时间至 72 h,凋亡率降至 ( 2 6.7± 1 .5) % .凋亡率与药物浓度的对数呈正相关 ( r=0 .938,P<0 .0 5) .结果表明 GP7具有诱导 NB4细胞凋亡作用 .

To explore the anti tumor mechanism of 4 〔4″ (2″,2″,6″,6″ tetramethyl 1″ piperidinyloxy)amino〕 4′ demethylepipodophyllotoxin(GP7) in apoptosis aspect, the growth inhibition effects of GP7 on leukemic NB4 cell line, the morphology of NB4 cell under light and electron microscope and the DNA ladder on agarose gel electrophoresis were observed. GP7 0.18-18 μmol·L -1 could markedly inhibit the growth and proliferation of NB4. GP7 induced apoptotic morphological changes were found under both light and electron microscopes and the “ladder” was observed by agarose gel electrophoresis. Apoptosis rate increased as time prolonged. The peak of apoptosis rate (45.0±3.0)% was reached at 48 h when NB4 was being exposured to GP7 9 μmol·L -1 . Apoptosis rate decreased to (26.7±1.5)% with prolonged exposure time to 72 h. There was a correlation between apoptosis rates and logarithmic GP7 concentration (r=0.938, P<0.05). We for the first time found that GP7 could induce NB4 apoptosis and the induction of apoptosis may be one of the anti tumor mechanisms of GP7.