小分子肽CSNRDARRC与膀胱移行细胞癌组织特异性结合的研究

Research on the specific combination of CSNRDARRC and bladder transitional cell carcinoma

目的明确小分子肽CSNRDARRC与膀胱移行细胞癌组织结合的特异性及其结合位点。方法选用膀胱移行细胞癌、肾癌、胃癌及腺性膀胱炎石蜡组织块,分别为107、43、68、16例;进行连续切片、烤片、脱蜡、抗原修复,封闭,再采用免疫荧光技术加FITC-六氨基己酸-CSNRDARRC复合物,荧光显微镜定性分析,单纯加FITC作为阴性对照。采用SPSS13.0软件进行统计分析,P<0.05为有显著性统计学差异。结果单纯FITC标记的各蜡块组织(阴性对照组)均未见有高亮荧光点,FITC-六氨基己酸-CSNRDARRC复合物标记的膀胱移行细胞癌组织有99例出现高亮荧光点,特异性为92.52%(99/107),而在肾癌、胃癌及腺性膀胱炎组织分别有6例、5例、2例有荧光亮点,特异性分别为9.52%(5/43),8.82%(6/68),12.50%(2/16),通过DAPI着色确定高亮荧光点主要定位于细胞核。CSNRDARRC与膀胱移行细胞癌组织的结合较其与肾癌、胃癌及腺性膀胱炎组织的结合有显著差异(P<0.05),而CSNRDARRC与肾癌、胃癌及腺性膀胱炎组织间结合比较无统计学意义(P>0.05)。结论小分子肽CSNRDARRC与膀胱移行细胞癌特异性结合率显著高于其他肿瘤组织,但对其他组织仍有标记,且结合位点在细胞核内,这为该肽段对膀胱移行细胞癌临床诊断提供理论基础;为膀胱癌的治疗奠定一定的理论依据。

Objective To explore the specificity of combination of peptide CSNRDARRC and bladder transitional cell carcinoma（TCC） tissues and the binding sites.Methods 107,43,68 and 16 paraffin-embedded tissues of TCC,renal carcinoma,gastric cancer and cystitis glandular were selected as Objectives.The tissues were sliced,baked,de-paraffined,antigen retrieved and blocked.After that,the tissues were mixed with FITC-six amino caproic acid-CSNRDARRC compound with immunofluorescence technique.The Results were analyzed qualitatively with fluorescence microscope.The negative controls were only mixed with FITC.The data were analyzed with statistical software SPSS13.0.Results No negative controls showed highlighted fluorescence points,while 99 cases of bladder TCC mixed with FITC-six amino caproic acid-CSNRDARRC compound showed highlighted fluorescence points and the specificity of combination was 92.52%（99/107）.6,5 and 2 cases of renal carcinoma,gastric cancer and cystitis glandular tissues respectively showed highlighted fluorescence points.The specificity of combination was 9.52 %（5/43）,8.82 %（6/68） and 12.50%（2/16）.The highlighted fluorescence points were located in nucleus by DAPI staining.Combination of CSNRDARRC with bladder TCC was of significant difference（P0.05）,while that with renal carcinoma,gastric cancer and cystitis glandular tissues was of no significant difference.Conclusions Combination of peptide CSNRDARRC with bladder TCC has higher combination ratio compared with other tumors,and the binding site of peptide CSNRDARRC is in the nucleus.These findings provide theoretical basis for the diagnosis of bladder TCC and the treatment of bladder carcinoma.