Specificity and Targeting of Optical Molecular Probe FITC-CSPLNTRFC on Breast Cancer Cells Bcap-37
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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:ObjectiveTo investigate the specificity and targeting abilities of FITC-CSPLNTRFC peptide FITC-BCSP1 optical molecular probe on breast cancer cells Bcap-37.
MethodsProbe 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.
ResultsThe 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.
ConclusionThe 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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Key words:
- Breast cancer /
- Targeting peptide /
- Optical molecular probe /
- Specificity /
- Targeting abilities
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Competing interests: The authors declare that they have no competing interests.作者贡献郭超：设计及实施实验、采集并分析数据、撰写论文左琪：设计及实施实验、采集并分析数据杨晓峰：设计及指导实施实验及论文修改张帆：设计及实施实验、获取研究经费、指导及审阅论文
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图 1 FITC-BCSP1探针与多种细胞结合的流式细胞图
Figure 1 Flow cytometry analysis of combination of FITC-BCSP1 probe and different cells
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图 2 光学分子探针与Bcap-37细胞结合的荧光倒置显微镜成像图
Figure 2 Fluorescence inverted microscope images of combination of optical molecular probes and Bcap-37 cells
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图 3 FITC-BCSP1探针吸光度值与浓度的对应关系(A)和最适浓度的光照稳定性(B)
Figure 3 Correspondence between absorbance and concentration of FITC-BCSP1 probe(A) and light stability of the optimum concentration(B)
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图 4 FITC-BCSP1探针注入荷瘤裸鼠模型体内3h时离体器官及肿瘤组织的荧光分子成像
Figure 4 Fluorescence molecular imaging of in vitro organs and tumor tissues after FITC- BCSP1 probe was injected into tumor-bearing nude mice for 3h
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图 5 FITC-BCSP1探针注入后3h肿瘤区荧光分子成像
Figure 5 Fluorescence molecular imaging of tumor region after FITC-BCSP1 probe was injected into tumor-bearing nude mice for 3h
表 1 不同浓度的FITC-BCSP1和FITC-svBCSP1探针对Bcap-37细胞的相对抑制率
Table 1 Inhibition rates of Bcap-37 cells by different concentrations of optical molecular probes FITC-BCSP1 and FITC-svBCSP1
下载: 导出CSV
表 2 光学分子探针标记的各类细胞百分比(x±s)
Table 2 Percentage of cells labeled by optical molecular probe (x±s)
下载: 导出CSV
表 3 肿瘤组织12 h内在不同时间荧光分子成像的灰度值
Table 3 Gray value of fluorescence molecular imaging of tumor tissues at different time within 12 h
下载: 导出CSV
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