CAJ | 학술논문

目的:通过研究钾离子通道阻断剂4-氨基吡啶(4-AP)对肿瘤化疗药物多西紫杉醇(docetaxel,DOC)的抗人乳腺癌细胞MCF-7作用的影响,探讨4-AP能否增强DOC的作用.方法:用MTT比色法分析DOC、4-AP以及两药联合应用对人乳腺癌细胞MCF-7增殖的影响;用流式细胞术PI单染法及Annexin V-FITC/PI双染法检测上述两种药物对人乳腺癌细胞MCF-7的细胞周期及早期凋亡的影响.结果:Dec能明显抑制人乳腺癌细胞株MCF-7的增殖,并呈剂量和时间依赖性.4-AP时MCF-7细胞增殖亦具有一定的抑制作用,在用药后24h、48h及72h的抑制卒分别为11.9%±1.7%、42.1%±3.2%、44.2%±1.6%.且5mmol/L4-AP可使DOC的作用增强,使25μmol/L DOC对人乳腺癌细胞株MCF-7最大杀伤作用时间从72h提前至24h.5μmol/L DOC能够使MCF-7G_2/M期细胞比率明显增加(53.58%±1.44%与对照组8.83%±0.44%相比,P<0.01),使G_0/G_1期细胞减少(11.48%±0.14%与对照组63.89%±0.98%相比,P<0.01),说明DOC主要在G_2/M期阻滞MCF-7细胞的增殖.5mmol/L 4-AP作用于MCF-7使其G_0/G_1期细胞比率增加,G_2/M期细胞明显减少,甚至消失(0.42%±0.17%与对照组8.83%±0.44%相比,P<0.05).而两药联用可见4-AP使MCF-7 G_2/M期细胞比率有所减少,而G_0/G_1期细胞比率有所增加.DOC单独作用于人乳腺癌细胞株MCF-7细胞PAh后,能明显增加晚期凋亡和死亡率(由6.97%±0.75%增加到20.77%±0.75%,P<0.05);而两药联合时,与时照组相比,早期凋亡比例由4.60%±0.91%增加到12.20%±0.82%(P<0.05),晚期凋亡和死亡细胞比例由6.97%±0.75%增加到58.42%±0.31%(P<0.05),提示4-AP(5mmol/L)能明显增加DOC诱导的MCF-7早期凋亡.结论:DOC和4-AP分别在G_2/M期和G_0/G_1期抑制MCF-7细胞增殖,4-AP通过促进DOC诱导细胞凋亡发挥抑制MCF-7细胞增殖的作用.
목적:통과연구갑리자통도조단제4-안기필정(4-AP)대종류화료약물다서자삼순(docetaxel,DOC)적항인유선암세포MCF-7작용적영향,탐토4-AP능부증강DOC적작용.방법:용MTT비색법분석DOC、4-AP이급량약연합응용대인유선암세포MCF-7증식적영향;용류식세포술PI단염법급Annexin V-FITC/PI쌍염법검측상술량충약물대인유선암세포MCF-7적세포주기급조기조망적영향.결과:Dec능명현억제인유선암세포주MCF-7적증식,병정제량화시간의뢰성.4-AP시MCF-7세포증식역구유일정적억제작용,재용약후24h、48h급72h적억제졸분별위11.9%±1.7%、42.1%±3.2%、44.2%±1.6%.차5mmol/L4-AP가사DOC적작용증강,사25μmol/L DOC대인유선암세포주MCF-7최대살상작용시간종72h제전지24h.5μmol/L DOC능구사MCF-7G_2/M기세포비솔명현증가(53.58%±1.44%여대조조8.83%±0.44%상비,P<0.01),사G_0/G_1기세포감소(11.48%±0.14%여대조조63.89%±0.98%상비,P<0.01),설명DOC주요재G_2/M기조체MCF-7세포적증식.5mmol/L 4-AP작용우MCF-7사기G_0/G_1기세포비솔증가,G_2/M기세포명현감소,심지소실(0.42%±0.17%여대조조8.83%±0.44%상비,P<0.05).이량약련용가견4-AP사MCF-7 G_2/M기세포비솔유소감소,이G_0/G_1기세포비솔유소증가.DOC단독작용우인유선암세포주MCF-7세포PAh후,능명현증가만기조망화사망솔(유6.97%±0.75%증가도20.77%±0.75%,P<0.05);이량약연합시,여시조조상비,조기조망비례유4.60%±0.91%증가도12.20%±0.82%(P<0.05),만기조망화사망세포비례유6.97%±0.75%증가도58.42%±0.31%(P<0.05),제시4-AP(5mmol/L)능명현증가DOC유도적MCF-7조기조망.결론:DOC화4-AP분별재G_2/M기화G_0/G_1기억제MCF-7세포증식,4-AP통과촉진DOC유도세포조망발휘억제MCF-7세포증식적작용.
Objective: To study the effect of docetaxet (DOC) combined with 4-AP on human breast can-cer MCF-7 cells and to explore whether 4-AP could strengthen the effect of docetaxel. Methods: MTT assays were performed to investigate the effect of docetaxel, 4-AP and the combination of them on the proliferation of MCF-7 cells. Flow cytometry was employed to detect cell cycles and cell apoptosis after the cells were stained by PI alone or by Annexin-V and PI. Results: Docetaxel could significantly inhibit the proliferation of MCF-7 cells in a dose- and time- dependent manner. 4-AP could inhibit the proliferation of MCF-7 cells and the inhibitory rates were 11.9%±1.7%, 42.1%±3.2%, and 44.2%±1.6% at 24h, 48h and 72h after adding 4-AP. Moreover 4-AP (5mmol/L) could strengthen the effect of docetaxel. 4-AP (25μmol/L) could increase the effect of Docetaxel. Docetaxel at 5μmol/L could significantly increase the percentage of cells at G_2/M (53.58%± 1.44% vs. 8.83%±0.44%, P<0.01) and decrease the percentage of cells at G_0/G_1 (11.48%±0.14% vs. 63.89%±0.98%, P<0.01), indicating that docetaxel blocked MCF-7 cells at G_2/M phase. 4-AP at 5mmol/L could in-crease the percentage of MCF-7 cells at G_0/G_1 and decrease the percentage of cells at G_2/M (0.42%±0.17% vs. 8.83%±0.44%, P<0.05). Docetaxel could significantly increase late apoptosis and death of MCF-7 cells af-ter treatment over 24h (from 6.97%±0.75% to 20.77%±0.75%, P<0.05). Docetaxel combined with 4-AP could increase early apoptosis rate from 4.60%±0.91% to 12.20%±0.82% (P<0.05) and could increase late apopto-sis rate and death rate from 4.60%±0.91% to 12.20%±0.82% (P<0.05). Conclusion: Both docetaxel and 4-AP can inhibit the proliferation of MCF-7 cells. Docetaxel can increase the percentage of cells at G_2/M phase and 4-AP can increase the percentage of cells at G_0/G_1 phase. 4-AP could strengthen the inhibitory effect of docetaxel on the proliferation of MCF-7 cells through inducing cell apoptosis.