摘要
目的:以人鼻咽癌CNE-1、CNE-2细胞为研究对象,采用激光共聚焦显微镜动态检测细胞对大黄素的摄取,探讨不同微环境对CNE-1、CNE-2细胞摄取大黄素的影响。方法:通过MTT方法检测大黄素对CNE-1和CNE-2细胞在乏氧和正常培养的条件下的生长抑制作用,选择无细胞毒性浓度的大黄素,激光共聚焦显微镜完美对焦系统分别观察两种细胞在乏氧、无血清、与血管内皮细胞共培养后对大黄素的摄取及这些过程的细胞形态学变化。结果:鼻咽癌2种细胞无论单独培养和共培养,在乏氧及无血清条件下摄取大黄素后,与相应的对照组细胞比较荧光强度最强,达到最大摄取浓度所需时间最短(P<0.05);总体而言,CNE-2细胞对大黄素的摄取速度快于CNE-1细胞,共培养体系中CNE-1细胞对大黄素的摄取速度慢于单细胞培养体系,而CNE-2细胞则相反。乏氧状态的鼻咽癌细胞在摄取大黄素后会出现明显的细胞皱缩、变圆、起泡等凋亡的形态学变化。大黄素对鼻咽癌CNE-1和CNE-2细胞在乏氧和不乏氧情况下半数抑制浓度(IC50)分别为18.34、16.87μg·mL-1和20.44、19.87μg·mL-1。结论:鼻咽癌细胞对大黄素的摄取与细胞的培养条件、乏氧状态等微环境有关,改变微环境有利于提高药物的靶向性。
Objective：To detect the emodin uptake of nasopharyngeal carcinoma cells CNE - 1 and CNE - 2 by eonfocal laser scanning microscopy and discuss the effect of different microenvironments on the emodin uptake of CNE - 1 and CNE - 2 . Methods： The growth inhibition of emodin to CNE - 1 and CNE - 2 cells under the condi- tion of hypoxia and normal cultivation was detected by the MTY assay. A non - cytotoxic concentration of emodin was used in the experiment. The emodin uptake of CNE - 1 and CNE - 2 cells that were cultivated under the condi- tion of hypoxia, serum - free and co - culture with vascular endothelial cells, and morphological changes were recor- ded by confocal laser scanning microscopy with a perfect focusing system. Results：After the emodin uptake under hypoxia and serum - free condition, regardless of mono - culture or co - culture, the fluorescence peak intensity of the nasopharyngeal carcinoma cells was stronger and the time to reach maximum concentration was shorter （P 〈 0. 05）, compared with those of the corresponding control cells. In general, the emodin uptake speed of CNE - 2 cells was faster than that of CNE - 1 cells . The emodin uptake speed of CNE - 1 cells in the co - culture system was slower than in the mono - culture system ; however, this situation was opposite to that of CNE - 2 cells . After the emodin uptake in the hypoxia environment, the cells would show morphologically apoptotic changes obviously such as cell pyknosis, becoming round and bubbling. Under the hypoxia and normal circumstances, the IC50 of emo- din were 18.34μg·mL^-1and 20. 44μg·mL^-1 for CNE - 1 cells,and 16. 87 μg·mL^-1 and 19. 87μg·mL^-1 for CNE - 2 ceils, respectively. Conclusion： Emodin uptake of nasopharyngeal carcinoma cells is related to the micro- environment, such as condition of culture, and state of hypoxia, etc.. Changing the microenvironment is favorable for improving drug targeting.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2014年第3期485-489,共5页
Chinese Journal of Pharmaceutical Analysis
基金
国家自然基金项目(No.81060270)
广西医学科学实验中心开放基金(No.KFJJ2011-25)
关键词
激光共聚焦显微镜
鼻咽癌细胞
大黄素摄取
微环境动态分析
大黄素抗癌活性分析
confocal laser scanning microscope
nasopharyngeal carcinoma cells
emodin uptake
microenvironmentdynamic observation
emodin antitumor activity analysis
