链霉亲和素修饰的量子点联合生物素标记的叶酸在卵巢癌SKOV3细胞体外成像中的应用

发布时间：2018-08-24 18:53

【摘要】：目的 :探讨链霉亲和素(streptavidin,SA)修饰的量子点(quantum dots,QDs)联合生物素标记的叶酸(folic acid,FA)在卵巢癌SKOV3细胞体外成像中的应用价值。方法 :采用活泼酯法合成SA-QDs复合物,并用同样的方法以聚酰胺-胺(polyamidoamine,PAMAM)树状分子为载体合成生物素化FA-PAMAM。采用生物素化FA-PAMAM联合SA-QDs荧光探针标记叶酸受体(folate receptor,FR)高表达的SKOV3细胞,同时以采用游离FA预处理的SKOV3细胞和FR低表达的肺腺癌A549细胞为对照,验证该荧光探针靶向SKOV3细胞的特异性。随后,以未用PAMAM为载体[生物素化FA-聚乙二醇(polyethylene glycol,PEG)联合SA-QDs荧光探针]或无生物素-链霉亲和素放大系统的荧光探针(FA-PAMAM-QDs)为对照,验证该探针的荧光信号双重放大效应。结果 :FR高表达的SKOV3细胞可被生物素化FA-PAMAM联合SA-QDs的荧光探针特异性识别,平均荧光强度高达114.92±2.87,显著高于FA预处理后再用该探针进行检测的SKOV3细胞(57.86±7.59)和FR低表达的肺腺癌A549细胞(14.94±0.83)(P值均0.000 1)。在双重放大效应的验证实验中,生物素化FA-PAMAM联合SA-QDs荧光探针标记的SKOV3细胞的平均荧光强度为120.89±3.80,其荧光强度明显高于生物素化FA-PEG联合SA-QDs荧光探针标记的SKOV3细胞的77.50±2.43和FA-PAMAMQDs荧光探针标记的SKOV3细胞的47.81±1.35(P值均0.000 1)。结论 :构建获得了一种特异性靶向卵巢癌SKOV3细胞的生物素化FAPAMAM联合SA-QDs荧光探针,该探针在卵巢癌早期诊断方面具有潜在应用价值。
[Abstract]:Aim: to investigate the value of streptavidin (streptavidin,SA) modified quantum dots (quantum dots,QDs) combined with biotin labeled folic acid (folic acid,FA) in the imaging of ovarian cancer SKOV3 cells in vitro. Methods: SA-QDs complex was synthesized by active ester method, and biotinylated FA-PAMAM. was synthesized by the same method using polyamide-amine (polyamidoamine,PAMAM) dendrimer as carrier. Biotin FA-PAMAM and SA-QDs fluorescence probe were used to label SKOV3 cells with high expression of folate receptor (folate receptor,FR), while SKOV3 cells pretreated with free FA and A549 cells with low expression of FR were used as control. The specificity of the fluorescent probe targeting SKOV3 cells was verified. Then, the double amplification effect of PAMAM was verified by using PAMAM as carrier [biotinylated FA- polyethylene glycol (polyethylene glycol,PEG) combined with SA-QDs fluorescent probe] or biotin-streptavidin amplification system (FA-PAMAM-QDs) as control. Results SKOV3 cells with high expression of 10% FR could be specifically recognized by the fluorescent probe of biotinylated FA-PAMAM combined with SA-QDs. The average fluorescence intensity was 114.92 卤2.87, which was significantly higher than that of SKOV3 cells (57.86 卤7.59) and A549 cells (14.94 卤0.83) (P, 0.000 1). In the double amplification experiment, The average fluorescence intensity of SKOV3 cells labeled with biotinylated FA-PAMAM and SA-QDs probe was 120.89 卤3.80, which was significantly higher than that of SKOV3 cells labeled with biotinylated FA-PEG and SA-QDs probe (77.50 卤2.43) and FA-PAMAMQDs labeled SKOV3 cells (47.81 卤1.35, P = 0.000 1). Conclusion: a biotinylated FAPAMAM combined with SA-QDs fluorescence probe targeting ovarian cancer SKOV3 cells was constructed. The probe has potential application value in early diagnosis of ovarian cancer.
【作者单位】：南昌大学第二附属医院妇产科;南昌大学食品科学与技术国家重点实验室;
【基金】：国家自然科学基金资助项目(编号:81201691) 高等学校博士学科点专项科研基金资助项目(编号:20123601120005) 江西省科技计划资助项目(编号:2015BBG70077)~~
【分类号】：R737.31

【参考文献】