Experimental study on the characters of the FITC-CSNRDARRC molecular probe and its targeted ability for orthotopic transplanted tumor of bladder cancer in vivo / 中华泌尿外科杂志

Objective To investigate the characters and targeted ability of FITC-CSNRDARRC molecular probe in labeling orthotopic transplantation tumor of bladder cancer in vivo.Methods From July 2013 to June 2014,the stability and characters of FITC-CSNRDARRC molecular probe were detected by spectrophotometer and molecular imaging and the optimum concentration and imaging time window were determined.30 BALB-C nude mice were randomly divided into experimental group (n =20) and control group(n =10).In control group,5 of them (group A) were ligated bilateral ureter,others(group B) were not.We established orthotropic transplanted bladder tumor (BIU-87) model by operation.And 0.2 ml probes (220 μmol/L) was then injected intravenously in all mice after 2 weeks.We obtained images and analyzed average gray value of the heart,lung,liver,spleen,bilateral kidney and orthotropic transplantation bladder tumor by using optical probe molecule fluorescence imaging system after 30 min,1 h,2 h,4 h and 12 h,respectively.Results After injected the FITC-CSNRDARRC molecular probes intravenously at 220 μmol/L,the fluorescence signal of tumor tissue strengthened gradually.The optimal imaging time window was 4 hours after injection.The illumination and temperature had little effect on the fluorescence signal.With the time passing after injection,the intensity of florescence signal progressively increasing,which reached the peak at4 h.The average gray value of tumor tissue at 1 h,2 h,3 h,4 h,5 h,6 h,8 h,12 h were 74.22,76.2,80.11,89.38,83.29,85.1,81.22,83.01,respectively.The fluorescence signal of normal tissue weakened gradually with the passage of time.Only liver and gall bladder could notice the fluorescence signal 4 hours after injection in group A.However,the relatively strong fluorescence signal could be found in liver and gall bladder in group B.Conclusions The characters of fluorescence probe are affected by its concentration.Its optimal concentration of labeling tumor is 220 μmol/L.The optical imaging time window was about 4 h after intravenous injection.The FITC-CSNRDARRC molecular probe can specifically bound to orthotopic transplanted tumor of bladder cancer in vivo.

Preclinical study of bladder cancer diagnosis using tumor homing peptide fluorescent molecular probe / 中华泌尿外科杂志

Objective To evaluate the features of optical molecular imaging of bladder tumor cells labeled by tumor homing peptide fluorescent molecular probe, and to explore the theoretical foundation of optical molecular imaging for bladder cancer diagnosis.Methods After prepared the FITC-CSNRDARRC fluorescent molecular probe, laser scanning confocal microscope, immuno fluorescence and multispectral fluorescence in vivo optical molecular imaging system have been used to evaluate the binding sites, the affecting factors of binding rates, the specificity and the targets.BIU-87 bladder tumor cell line, BIU-87 bladder tumor cell line, 68 cases of paraffin bladder tumor tissue samples, 16 cases of paraffin glandular bladder inflammatory samples, 43 cases of paraffin renal clear cell carcinoma samples, 68 cases of paraffin gastric adenocarcinoma samples, 29 cases of urine exfoliated cells suspected bladder cancer and BIU-87 bladder cancer nude xenograft have been used in this study.Results The binding site of FITC-CSNRDARRC fluorescent molecular probe were at the nucleus of labeled bladder tumor cells.The binding rates were correlated linearly with the dose of probe and the grade of pathology.The in vitro and in vivo studies demonstrate that the probe has a binding specificity with bladder tumor. When the FITC-CSNRDARRC fluorescent molecular probe labeled tumor cells, bright green spots were observed under laser scanning confocal microscope.The bright green spots were more apparent after stained by DAPI again.The tissue samples and tumor cells in the urine can be successful labeled and identified by fluorescence microscope.Optical molecular imaging of in vivo xenograft tumor tissues showed fluorescent spots under EMCCD.Conclusions The labeled loci of single cell by FITC-CSNRDARRC probe have been identified. The spatial resolution of optical molecular image is related to sensitivity of CCD, and the optical molecular imaging cannot be imaged by the conventional endoscope camera.

Selection of peptide specifically binding to bladder carcinoma by using phage display in vivo / 中国免疫学杂志

Objective:To screen the peptide binding to human bladder carcinoma cells specifically by using phage display technology in vivo.Methods: Nude mice were inoculated with bladder carcinoma cells BIU87 for establishing tumor-bearing mice model.The Ph.D.-C7CTM Peptide Library was injected intravenously via tail vein.Then we screened Phage containing exogenous peptides binding to bladder transitional carcinoma cells specifically.The phage peptide homed to the tumor tissues was obtained after 3 rounds screening in vivo.The phage clones affinity to BIU87 were identified by immunohistochemistry and ELISA.The positive peptide was synthetized by chemical methods after sequencing the positive monoclonal phage DNA.The tumor cell specificity of target peptide was identified by confocal laser scanning microscope and flow cytometry.Results:After 3 rounds screening in vivo,enrichment rate of phage was 4.334×102 times.Immunohistochemistry results showed that the dyeing of the tumor tissue had a rising trend following each round of phage screening,while liver had a lot of non-specific binding phage because the phages were metabolized through liver and kid-ney.The 30 phage clones were identified by ELISA and 10 clones had a strong affinity on BIU87 among 24 positive clones.Three amino acid sequences of positive phage clones were obtained.The highest rate of repeat sequences CSSPIGRHC(8/10) named NYZL1 and the FITC-C6-NYZL1 peptide was synthesized.Our results showed that it could bind to bladder carcinoma cells BIU87 specifically.Conclusion:We obtained the small molecular peptide NYZL1 binding to human bladder carcinoma specifically by means of phage display in vivo,which provide a theoretical basis for bladder carcinoma early diagnosis and targeted therapy.

Advances of peptides in cancer treatment / 国际肿瘤学杂志

Studies demonstrate that peptides have great potential in cancer therapy and inhibitation of tumor progression.Peptides can be used as hormones,vaccines,carriers of redionuclides and cytotoxic drugs as well as anti-tumor drugs playing important roles in oncotherapy.Targeting chemotherapy and targeting drug transportation technique can enable the drugs to concentrate expected location selectively and effectively,emerging as an alternative to the conventional chemotherapy.