基于特异性结合多肽ZS-1的肺癌新肿瘤标志物的筛选

Screening of tumor mark for the lung cancer by using peptide ZS-1

目的:利用本实验室已获得的与肺癌特异性结合的十二肽ZS-1(EHMALTYPFRPP)从人类肺癌cDNA文库中筛选出肺癌相关标志物,并进行初步确认,为肺癌的临床早期诊断和靶向治疗奠定基础。方法:本实验以ZS-1为探针,通过将偶联生物素的多肽ZS-1固化在亲和素处理的酶标板,筛选人类肺癌cDNA噬菌体展示文库,经过三轮减性筛选后,挑取与ZS-1特异性结合的噬菌体单克隆、扩增、纯化,再通过PCR技术扩增、产物纯化后,进行DNA测序,测序结果经过生物信息学分析,随后采用组织芯片技术对相关生物大分子进行了检测。结果:通过生物信息学分析首次发现与ZS-1结合的肺癌细胞膜受体蛋白为KIAA1199,随后以KIAA199抗体检测人肺癌组织芯片证实,KIAA1199的阳性检出率为32.8%。结论:从人类肺癌cDNA噬菌体展示文库中筛选出潜在的肺癌肿瘤标志物KIAA1199,为肺癌的早期诊治奠定基础。

AIM：To select a potential biomarker for early lung cancer diagnosis and targeted therapy by using the cancer specific bounded peptide ZS-1 which had already been obtained from the laboratory. METHODS：The peptide ZS-1 marked by biotin was used as a probe to pan out the human lung cancer cDNA phage display library,after 3 rounds of subtraction panning,the specific binding clones of ZS-1 were identified.After amplification and purification,then those DNA sequences were identified and analyzed with bioinformatics.Tissue microarray technology was used to detect biomelecule in tissues.RESULTS：The lung cancer membrane receptor protein KIAA1199 binding ZS-1 was first found by bioinformatics analysis.Subsequently,a specific human lung cancer tissue microarray was detected and analysed by KIAA199 antibody.The results displayed KIAA1199 positive detection rate was 32.8%.CONCLUSION：A tumor biomarker KIAA1199 selected from human lung cancer cDNA library provides a potential tool for early lung cancer diagnosis and therapy.