目的 制备精氨酸-甘氨酸-天冬氨酸(RGD)与核糖核酸酶(RNase A)修饰的碲化镉(CdTe)量子点(quantum dot,QDs)的纳米探针,观察其对恶性黑素瘤A375细胞的靶向效果.方法 利用微波加热方法得到核糖核酸酶修饰的碲化镉量子点(CdTe RQDs),再化学键合偶联RGD多肽得到RGD-CdTe RQDs纳米探针,通过透射电镜、粉末晶体衍射、荧光光谱仪和紫外吸收光谱仪检测其相应物理和光学表征.体外培养A375细胞,通过SPSS软件统计分析MTT实验结果,确定用于细胞成像的RGD-CdTe RQDs探针浓度,与A375细胞共同孵育15 min,通过激光共聚焦显微镜荧光成像的结果研究RGD-CdTe RQDs纳米探针与A375细胞之间结合的特异性.结果 用微波加热方法制备分散性和生物相容性好的CdTe RQDs纳米探针,通过化学偶联成功构建RGD-CdTe RQDs纳米探针,MTT实验结果表明,用20、40、80 nmol/L的RGD-CdTe RQDs探针与A375细胞孵育12、24、36和72 h后,20 nmol/L的RGD-CdTe RQDs在12h内对A375细胞的生命活动影响最低;选择20 nmol/L的RGD-CdTe RQDs进行荧光成像实验,发现偶联RGD多肽的CdTe RQD纳米探针对A375细胞有明显的主动靶向效果.结论 成功制备RGD-CdTe RQDs纳米探针,该荧光分子探针可以主动靶向A375细胞.
Objective To prepare arginine-glycine-aspartic acid (RGD)-and ribonuclease A (RNase A)-conjugated CdTe quantum dot (QD) nanoprobes,and to observe their capability to target human A375 malignant melanoma cells.Methods RNase A-modified CdTe quantum dots (CdTe RQDs) were obtained by using a microwave-based heating method,and then chemically conjugated to the RGD peptide to prepare RGD-CdTe RQD nanoprobes,which were then physically and chemically characterized by transmission electron microscopy,powder crystal diffraction,fluorescence spectrophotometry,and ultraviolet absorption spectrophotometry.A375 cells were cultured in vitro and incubated with various concentrations (20,40,80 nmol/L) of RGD-CdTe RQD nanoprobes for different durations (12,24,36,72 hours).Then,methyl thiazolyl tetrazolium (MTT) assay was conducted to estimate the proliferative activity of A375 cells.To observe the targeting capability of RGD-CdTe RQD nanoprobes,A375 cells were treated with RGD-CdTe RQD nanoprobes at the