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光学分子探针FITC-CSPLNTRFC对乳腺癌细胞Bcap-37的特异性和靶向性

Specificity and Targeting of Optical Molecular Probe FITC-CSPLNTRFC on Breast Cancer Cells Bcap-37

  • 摘要:
    目的 研究光学分子探针FITC-BCSP1对乳腺癌细胞Bcap-37的特异性和靶向性。
    方法 固相合成法制备探针FITC-BCSP1和阴性对照探针FITC-svBCSP1。MTT法检测两种探针对乳腺癌Bcap-37细胞的毒性。流式细胞术和荧光倒置显微镜鉴定FITC-BCSP1与Bcap-37细胞结合的特异性。光学分子成像仪验证FITC-BCSP1对Bcap-37细胞荷瘤裸鼠模型移植瘤的特异性和靶向性。
    结果 质谱和高效液相色谱法鉴定,所合成探针纯度≥98%。探针FITC-BCSP1和对照探针FITC-svBCSP1在50~300 μmol/L浓度内对Bcap-37细胞增殖、活性无影响(IR%≤30%)。流式细胞术显示Bcap-37细胞中FITC-BCSP1标记的细胞百分比明显高于其他细胞(均P < 0.001);FITC-BCSP1标记的Bcap-37细胞百分比明显高于对照组(P < 0.001)。荧光倒置显微镜下观察到,FITC-BCSP1标记Bcap-37细胞中有大量荧光细胞,阳性率为100%,而FITC-svBCSP1组阳性率仅1%。Bcap-37细胞荷瘤裸鼠模型体内验证实验显示,FITC-BCSP1能特异性富集在移植瘤组织。
    结论 光学分子探针FITC-BCSP1有良好的乳腺癌细胞特异性及靶向性,可用于乳腺癌早期诊断研究。

     

    Abstract:
    Objective To investigate the specificity and targeting abilities of FITC-CSPLNTRFC peptide FITC-BCSP1 optical molecular probe on breast cancer cells Bcap-37.
    Methods Probe FITC-BCSP1 and negative control probe FITC-svBCSP1 were prepared by solid phase synthesis. MTT assay was used to determine the toxicity of the two probes on breast cancer cells Bcap-37. The specificity of the binding of FITC-BCSP1 probe to Bcap-37 cells was identified by flow cytometry and fluorescent inversed microscopy. The specificity and targeting abilities of FITC-BCSP1 probe for transplantation tumor in Bcap-37 cells tumor-bearing nude mice model were tested by optical molecular imager.
    Results The purity of the synthesized probe was more than 98%, identified by mass spectrometry and high performance liquid chromatography. FITC-BCSP1 and FITC-svBCSP1 probes had no effect on proliferation and activity of Bcap-37 cells at the concentrations of 50-300 mol/L (IR%≤30%). FCM results showed that the percentage of FITC-BCSP1-labeled cells in Bcap-37 cells was significantly higher than that in other cells (all P < 0.001), and the percentage of FITC-BCSP1-labeled Bcap-37 cells was significantly higher than that of the control group (P < 0.001). It was observed under inverted fluorescence microscope that there were a large number of fluorescent cells in FITC-BCSP1-labeled Bcap-37 cells, with a positive rate of 100%, while the positive rate of FITC-svBCSP1 group was only 1%. In vivo assay with Bcap-37 cells tumor-bearing nude mice model showed that FITC-BCSP1 probe could be specifically enriched in the transplantation tumor tissue.
    Conclusion The optical molecular probe FITC-BCSP1 has good specificity and targeting abilities on breast cancer cells and can be used in the early diagnosis of breast cancer.

     

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