摘要
[目的]建立流式细胞术分析肿瘤细胞内阿霉素分布的方法,并研究低浓度五味子乙素对K562细胞内化阿霉素的影响。[方法]体外培养猪内皮细胞(pEC),以20μmol/L阿霉素处理2、4和6小时;或加入不同浓度阿霉素(0、10、20、40、80和100μmol/L)处理4小时。培养K562细胞,以5μmol/L阿霉素处理2、4和6小时;或加入不同浓度阿霉素(0、2.5、5、10和20μmol/L)处理4小时。采用不同浓度(0、25、50和100μmol/L)五味子乙素(Sch B)与阿霉素联合处理K562细胞4小时。收集细胞,采用流式细胞术分析阿霉素的特异荧光。[结果]固定浓度处理pEC(20μmol/L)和K562(5μmol/L)后检测,发现细胞内荧光强度随时间延长而增加,两种细胞4小时组荧光强度分别达到6小时组的96.93%和95.23%/。在直方图上,阴性和阳性细胞群界限分明。阿霉素(2.5、5和10μmol/L)单独处理细胞的荧光强度分别为26.78±3.34、64.70±6.24和118.35±9.67;添加25μmol/L Sch B实验后荧光强度增加到43.45±4.25、103.74±7.36和146.69±8.32,Sch B显著增加了K562细胞内阿霉素的分布(p<0.05)。[结论]建立的方案可快速测定细胞内的阿霉素分布;低浓度Sch B促进肿瘤细胞内化阿霉素。
Objective： To establish a method to analyze content of adriamycin in tumor cells by flow cytometry（FCM） and to analyze the effect of internalization of adriamycin into tumor cells promoted by schisandrin B.Method： Pig endothelia cells（pEC） were cultured with or without 20μmol/L adriamycin for indicated time（2,4 and 6 hours）,or in the presence of different concentration（0,10,20,40 and 80μmol/L） of adriamycin for 4 hours.Human erythroleukemia cells K562 were cultured with or without 5μmol/L adriamycin for indicated time（2,4 and 6 hours）,or in the presence of different concentration（0,2.5,5,10 and 20μmol/L） of adriamycin for 4 hours.K562 cells were treated by different concentration of adriamycin in the presence of schisandrin B（0,25,50 and 100 5μmol/L）.The specific red fluorescence of adriamycin in those cells were detected by FCM.Result： The fluorescence in pEC and K562 cells treated by determined concentration of adriamycin（10μmol/L or 5μmol/L） increased time-dependently.Compared with cells treated for 6 hours,cells treated for 4 hours presented almost 96.93% and 95.23 of fluorescence.Fluorescence positive and negative cells were distinct well in the histograms.The fluorescence in K562 cells treated by adriamycin（2,5 and10 5μmol/L） along were 26.78±3.34、64.70±6.24 and 118.35±9.67.The addition of 25μmol/L Sch B promoted the internalization of adriamycin to 43.45±4.25、103.74±7.3 and 146.69±8.32（p0.05）.Conclusion： The method to analyze content of adriamycin in tumor cells by FCM was established.It was confirmed that lower concentration of Sch B could promoted the internalization of adriamycin.
出处
《中国科技信息》
2012年第7期151-152,共2页
China Science and Technology Information
基金
吉林省教育厅十二五科研基金(编号2011420)
