量子点对膀胱癌BIU-87细胞抗原的标记检测

Application of Quantum-Dot-Based Immunofluorescence for the Special Detection of Bladder Cancer Cell Line BIU-87

目的:探讨不同的量子点免疫荧光组织化学方法对膀胱癌BIU-87细胞抗原特异性标记的效果,以优化细胞标记方法。方法:分别采用量子点免疫荧光组织化学三步法、二步法、一步法检测前列腺干细胞抗原(PSCA)在膀胱癌BIU-87细胞中的表达情况,观察并分析三种方法特异性标记的效果。结果:PSCA特异性的表达于BIU-87细胞胞质或膜,三种方法在紫外光激发下均能观察到表达部位明显的红色荧光,三步法实验技术成熟,量子点标记的链霉亲和素性质稳定,检测灵敏度高,背景清晰,但实验步骤繁琐,耗时较长,不宜用于活细胞标记;二步法实验步骤和操作时间都相对有所减少,但量子点标记的二抗性质不稳定,非特异吸附较强;一步法操作简便,省时,标记效果明显,特异性强,但是标记一抗成本较高,且对一抗的剂量要求较大。结论:三种量子点荧光标记方法均能对固定细胞表面抗原进行特异性标记,需根据不同实验需求选择合适的标记方法以达到最佳实验效果。特异性一步法量子点荧光标记在活细胞及活体靶向成像示踪中具有巨大的应用前景。

Objective：To investigate the effect of different quantum dots（QDs）based immunofluorescence histochemical methods for special detection of prostate stem cell antigen（PSCA）in the human bladder cancer cell line BIU-87,and optimize the method of cell labeling.Methods：The three-step,two-step and one-step method of quantum-dot-fluorescence labeling was used to detect the expression of PSCA in BIU-87 cells,respectively,and the effect of three kinds of specific labeling methods was observed and analyzed.Results：The obvious orange red fluorescence was observed in the cell cytoplasm and membrane by all of three kinds of methods.The three-step method had the characteristics of mature experiment technology,stable properties of QDs-streptavidin and the high detective sensitivity,and the background was clear,but the experimental process of this method was tedious and time consuming,so it was not appropriate for living cells labeling.The experimental steps and the operating time of two-step method were relatively decreased,however,QDs-biotinylated sheep-anti-rabbit was not stable and had a strong nonspecific adsorption.Finally,one-step method was simple,time-saving,and had an obvious labeled effect and strong specificity,nonetheless,the large doses of QDs and antibodies were needed when QDsconjugated antibodies,which would result in very high expenses.Conclusion：All the three methods of quantum-dot-fluorescence labeling can specifically mark the cell antigen,we should choose the appropriate labeling method according to different experimental demand in order to achieve the best effect of the experiment.One-step method of quantum-dot-fluorescence labeling has a great prospect in living cells and living targeted tracing.