CAJ | 학술논문

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.
목적 제비정안산-감안산-천동안산(RGD)여핵당핵산매(RNase A)수식적제화력(CdTe)양자점(quantum dot,QDs)적납미탐침,관찰기대악성흑소류A375세포적파향효과.방법 이용미파가열방법득도핵당핵산매수식적제화력양자점(CdTe RQDs),재화학건합우련RGD다태득도RGD-CdTe RQDs납미탐침,통과투사전경、분말정체연사、형광광보의화자외흡수광보의검측기상응물리화광학표정.체외배양A375세포,통과SPSS연건통계분석MTT실험결과,학정용우세포성상적RGD-CdTe RQDs탐침농도,여A375세포공동부육15 min,통과격광공취초현미경형광성상적결과연구RGD-CdTe RQDs납미탐침여A375세포지간결합적특이성.결과 용미파가열방법제비분산성화생물상용성호적CdTe RQDs납미탐침,통과화학우련성공구건RGD-CdTe RQDs납미탐침,MTT실험결과표명,용20、40、80 nmol/L적RGD-CdTe RQDs탐침여A375세포부육12、24、36화72 h후,20 nmol/L적RGD-CdTe RQDs재12h내대A375세포적생명활동영향최저；선택20 nmol/L적RGD-CdTe RQDs진행형광성상실험,발현우련RGD다태적CdTe RQD납미탐침대A375세포유명현적주동파향효과.결론 성공제비RGD-CdTe RQDs납미탐침,해형광분자탐침가이주동파향A375세포.
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.