国际肿瘤学杂志

国际肿瘤学杂志 ›› 2014, Vol. 41 ›› Issue (2): 144-147.doi: 10.3760/cma.j.issn.1673-422X.2014.02.019

• 论著 • 上一篇    下一篇

MDA-MB-231乳腺癌细胞特异性多肽的初步筛选

孔冕, 李宝江, 王军业, 曹开源, 林雨, 戴淑琴   

  1. 266021 青岛大学医学院（孔冕）；中山大学华南肿瘤学国家重点实验室（孔冕、王军业、戴淑琴），微生物教研室（曹开源）,高血压研究所（林雨）；271000 泰安市中心医院乳腺外科（李宝江）孔冕和李宝江对本文有同等贡献，均为第一作者
  • 收稿日期:2013-06-04 修回日期:2013-11-21 出版日期:2014-02-08 发布日期:2014-01-26
  • 通讯作者: 戴淑琴，E-mail: daishq@sysucc.org.cn E-mail:daishq@sysucc.org.cn
  • 基金资助:

    广东省科技计划项目（2010B031600066）

Screen peptides specifically bonding to MDA-MB-231 breast cancer cells

Kong Mian, Li Baojiang, Wang Junye, Cao Kaiyuan, Lin Yu, Dai Shuqin   

  1. Qingdao University Medical College; State Key Laboratory of Oncology in Southern China, Sun YatSen University, Qingdao 266021， China;#Department of Breast Surgery,Center Hospital of Tai′an, Tai′an 271000, China
  • Received:2013-06-04 Revised:2013-11-21 Online:2014-02-08 Published:2014-01-26
  • Contact: Dai Shuqin E-mail:daishq@sysucc.org.cn

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摘要： 目的 利用噬菌体展示技术，从噬菌体随机十二肽库中筛选出能够特异性结合MDA-MB-231乳腺癌细胞的噬菌体克隆。方法 以人正常乳腺细胞为减性筛选细胞、MDA-MB-231乳腺癌细胞为靶细胞，对噬菌体随机十二肽库进行筛选，挑取富集后的阳性单克隆噬菌体，酶联免疫吸附试验（ELISA）及DAB染色鉴定阳性噬菌体的特异性及亲和力。结果 经过3轮筛选，噬菌体得到约113倍的富集，随机挑选11株单克隆噬菌体，ELISA显示8号噬菌体单克隆对乳腺癌细胞的亲和力是对照的6.5倍，DAB鉴定亦显示其对乳腺癌细胞的特异性及亲和力最高，命名为LK-8。结论 利用噬菌体筛选技术成功筛选出能够特异性结合MDA-MB-231乳腺癌细胞的特异性噬菌体单克隆LK-8，可为进一步合成特异性多肽用于早期诊断和靶向治疗乳腺癌奠定基础。

关键词: 乳腺肿瘤, 酶联免疫吸附测定, 生物学标记, 噬菌体展示

Abstract: Objective To screen phage clones which can specific binding to MDA-MB-231 breast cancer cells from phage random 12 peptide library by phage display technology. Methods Agminated positive monoclonal phage was selected from phage random 12 peptide library, using normal mammary gland cells as subtract screening cells and MDA-MB-231 breast cancer cells as target cells. The specificity and affinity of the positive clones were identified by enzyme linked immunosorbent assay (ELISA) and DAB dyeing. Results Eleven clones were chosen after 3 rounds of screening. ELISA showed the affinity to breast cancer cells of NO.8 phage (named LK-8) was 6.5 times higher than that in control group, immunohistochemistry also showed NO.8 phage had a high specificity bonding to breast cancer cells. Conclusion A specific phage (LK-8) bonding to MDA-MB-231 breast cancer cells is screened successfully using phage display technology, and it lays the foundation for compounding specific polypeptide to diagnose and treat breast cancer.

Key words: Breast neoplasms, Enzyme linked immunosorbent assay, Biological markers, Phage display