中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (11): 800-804.

• 论著 • 上一篇    下一篇

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

陈晓罡1,2,张振3,费烨4,陈向东3   

  1. 1. 上海市皮肤病医院
    2. 上海市皮肤病医院 2号楼209手术室
    3. 上海交通大学医学院附属第九人民医院
    4. 上海市第九人民医院
  • 收稿日期:2012-12-04 修回日期:2013-07-05 出版日期:2013-11-15 发布日期:2013-11-01
  • 通讯作者: 陈向东 E-mail:xdchen@medmail.com.cn
  • 基金资助:
    国家自然科学基金面上项目

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

  • Received:2012-12-04 Revised:2013-07-05 Online:2013-11-15 Published:2013-11-01
  • Supported by:
    基金证明见投稿附件PDF

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摘要： 【摘要】 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12 h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。 【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

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

Abstract: CHEN Xiao-gang, ZHANG Zhen, FEI Ye, CHEN Xiang-dong*. *Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University, Shanghai 200011, China Corresponding author: CHEN Xiang-dong, Email: xdchen@medmail.com.cn 【Abstract】 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 concentration determined by MTT assay for one hour followed by laser confocal 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. 【Key words】 Melanoma; Quantum dots; Molecular probes; Protein transport; RGD

Key words: melanoma, Quantum dots

中图分类号: 

  • R751