精氨酸-甘氨酸-天冬氨酸与核糖核酸酶修饰碲化镉量子点探针对黑素瘤A375细胞的靶向研究  

Arginine-glycine-aspartic acid- and RNase A-conjugated CdTe quantum dot-based nanoprobes for active targeting of human A375 malignant melanoma cells in vitro

作　　者：陈晓罡 张振[1] 费烨[1] 陈向东[1] 

机构地区：[1]上海交通大学附属第九人民医院皮肤科,200011 [2]上海市皮肤病医院,200443

出　　处：《中华皮肤科杂志》2013年第11期800-804,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81272987）本文实验在上海交通大学微纳科学技术研究院纳米生物教研室完成

摘　　要：目的制备精氨酸-甘氨酸-天冬氨酸（RGD）与核糖核酸酶（RNaseA）修饰的碲化镉（CdTe）量子点（quantumdot，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（CdTeRQDs），再化学键合偶联RGD多肽得到RGD—CdTeRQDs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MTI＂实验结果，确定用于细胞成像的RGD—CdTeRQDs探针浓度，与A375细胞共同孵育15min，通过激光共聚焦显微镜荧光成像的结果研究RGD—CdTeRQDs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CdTeRQDs纳米探针，通过化学偶联成功构建RGD—CdTeRQDs纳米探针，MTT实验结果表明，用20、40、80nmoFL的RGD—CdTeRQDs探针与A375细胞孵育12、24、36和72h后，20nmol/L的RGD—CdTeRQDs在12h内对A375细胞的生命活动影响最低；选择20nmol／L的RGD—cdrreRQDs进行荧光成像实验，发现偶联RGD多肽的CdTeRQD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—CdTeRQDs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 （MTF） 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 concentration determined by MTI＂ assay for one hour followed by laser confoeal microscopy. Results CdTe RQDs with good dispersion and biocompatibility were obtained by using a microwave-based heating method, and then successfully conjugated to the RGD peptide to form RGD-CdTe RQD nanoprobes. The treatment with RGD-CdTe RQDs of 20 nmol/L for 12 hours exhibited the weakest effect on the proliferative activity of A375 cells, and hence, 20 nmol/L was selected for the fluorescence imaging assay. Laser confocal microscopy revealed that RGD-CdTe RQD nanoprobes were able to actively target A375 cells. Conclusion RGD-CdTe RQD nanoprobes with a favorable capability to actively target A375 cells are successfully prepared in this study.

关 键 词：黑色素瘤 量子点 分子探针 蛋白质转运 精氨酸-甘氨酸-天冬氨酸 

分 类 号：R739.5[医药卫生—肿瘤]