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| 肺癌噬菌体展示肽库的构建及肺癌早期检测分子标志的筛选 |
| 赵晋丰1,吴玲玲1,常文军1,黄盛东2,余永伟3,赵立军4,陈祎佳1,曹广文1* |
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| (1.第二军医大学卫生勤务学系流行病学教研室,上海 200433;2.第二军医大学长海医院胸心外科,上海 200433;3.第二军医大学长海医院病理科,上海 200433;4.第二军医大学长海医院呼吸内科,上海 200433) |
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| 摘要: |
| 目的:构建肺癌噬菌体展示肽库,筛选出肺癌早期检测的分子标志群。方法:取30例第二军医大学长海医院手术后立即冻存的肺癌组织标本构建肺癌噬菌体展示肽库；用结合 protein-A/G的琼脂糖珠分别富集肺癌组及非肿瘤对照组血浆中的抗体进行5轮亲和筛选,富集肺癌特异性的相关肽；随机挑取5轮筛选后的500个噬菌体克隆,用ELISA法进一步筛选肺癌/对照D比值大于2.0的肺癌特异标志物克隆,进行基因序列测定和蛋白功能预测。结果:(1)该噬菌体展示肽库的滴度为3.0×106 pfu,库容9.0×106 pfu,随机挑选20个噬菌斑经PCR鉴定其重组率为60%。(2)共筛选出19个有意义的噬菌体,测序后,预测其蛋白质功能,9个与癌症相关,8个功能未知,2个与癌症不相关。结论:筛选出的噬菌体克隆所表达的抗原很可能是早期诊断肺癌的标志物。 |
| 关键词: 噬菌体展示肽库 肺肿瘤 分子标志 |
| DOI：10.3724/SP.J.1008.2008.00 |
| 投稿时间：2008-04-02修订日期：2008-05-26 |
| 基金项目:上海市登山计划重大课题(06DZ19503). |
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| Screening for molecular markers for early diagnosis of lung cancer by bacteriphage display |
| ZHAO Jin-feng1,WU Ling-ling1,CHANG Wen-jun1,HUANG Sheng-dong2,YU Yong-wei3,ZHAO Li-jun4,CHEN Yi-jia1,CAO Guang-wen1* |
| (1. Department of Epidemiology,Faculty of Health Service,Second Military Medical University,Shanghai 200433,China;2. Department of Cardiothoracic Surgey,Changhai Hospital,Second Military Medical University,Shanghai 200433;3. Department of Pathology,Changhai Hospital,Second Military Medical University,Shanghai 2004334. Department of Respiratory Medicine,Changhai Hospital,Second Military Medical University,Shanghai 200433;4. Department of Respiratory Medicine,Changhai Hospital,Second Military Medical University,Shanghai 200433) |
| Abstract: |
| Objective:To establish a lung cancer phage-peptide library and to screen for biomarkers for early detection of lung cancer.Methods: A phage display library was constructed using 30 lung-cancer tissue samples from Changhai Hospital. Protein-A/G agarose was used to enrich IgG from control sera as well as lung cancer sera. Five biopannings were carried out for enrichment of lung cancer-specific phage clones. Five hundred phage clones were randomly selected,those with D>2.0( lung cancer plasma pool v.s. control plasma pool)were selected for DNA sequencing and protein prediction. Results: (1) The recombination rate of the phage library was 60%,with the average phage titer being 3.0×106 pfu and a volume being 9.0×106 pfu.(2) Of 19 phage colonies selected by ELISA and sequenced,9 were cancer-related genes,8 with unknown function,and 2 were not related to cancer. Conclusion: The screened genes in the phage colonies might serve as biomarkers for the early diagnosis of lung cancer. |
| Key words: phage-peptide library lung neoplasms biomarker |
