一种与前列腺癌高转移细胞表面受体特异性结合的多肽片段的筛选及验证

发布时间：2018-05-18 21:43

  本文选题：肽库 + 前列腺癌　； 参考：《肿瘤防治研究》2017年09期

【摘要】：目的利用细菌鞭毛肽库技术筛选与高转移潜能的前列腺癌PC-3M-1E8细胞特异结合的多肽片段。方法利用细菌鞭毛肽库对人前列腺癌高低转移配对细胞株PC-3M-1E8和PC-3M-2B4进行筛选,获得与前列腺癌高转移细胞亲和的细菌克隆。将结合能力最强细菌克隆所共同拥有的重复多肽序列挑选出来并化学合成;利用荧光染色和流式细胞仪,通过细胞亲和、竞争拮抗实验及荷瘤小鼠模型,鉴定合成肽与前列腺癌高转移细胞的体内外的结合特异性。结果筛选出一段核心序列为NVVRQ的五肽,命名为TMTP1;FITC-TMTP1可特异地与PC-3M-1E8高度亲和,且这种结合是浓度依赖并能为游离的TMTP1所拮抗;荧光显微镜检测小鼠肿瘤及正常组织器官冰冻切片显示FITC-TMTP1在肿瘤原发灶及转移灶的荧光信号强度也要显著强于正常组织器官。结论筛选获得的短肽TMTP1可以特异地与高转移潜能的前列腺癌细胞结合,为前列腺癌的靶向诊疗提供一种可能的标志物及靶向载体。
[Abstract]:Objective to screen polypeptide fragments specifically bound to prostate cancer PC-3M-1E8 cells with high metastatic potential by bacterial flagellin library technique. Methods Human prostate cancer cell lines PC-3M-1E8 and PC-3M-2B4 were screened by bacterial flagellin library, and bacterial clones with high affinity to prostate cancer cells were obtained. The repeated polypeptide sequences shared by bacterial clones with the strongest binding ability were selected and chemically synthesized. By means of cell affinity, competitive antagonism and tumor-bearing mouse models, fluorescence staining and flow cytometry were used. To identify the binding specificity of synthetic peptides to prostate cancer cells with high metastasis in vitro and in vivo. Results A pentapeptide with a core sequence of NVVRQ was selected and named TMTP1FITC-TMTP1, which was highly compatible with PC-3M-1E8, and the binding was concentration-dependent and antagonized by free TMTP1. Fluorescence microscopy showed that the fluorescence intensity of FITC-TMTP1 in primary tumor and metastatic tumor was significantly higher than that in normal tissue and organ. Conclusion the short peptide TMTP1 can specifically bind to prostate cancer cells with high metastatic potential and provide a possible marker and target vector for the diagnosis and treatment of prostate cancer.
【作者单位】： 华中科技大学同济医学院附属同济医院妇产科;
【基金】：国家自然科学基金(81202061,81572563)
【分类号】：R737.25
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本文编号：1907319