Construction and Identification of New Molecular Markers of Triple-Negative Breast Cancer Stem Cells

被引:1
作者
Liu, Tingting [1 ]
Wang, Hongyue [2 ]
Liu, Zhiyong [1 ]
Zhang, Jing [3 ]
Liu, Yan [1 ]
Zhang, Lin [1 ]
Zheng, Chunhui [4 ]
Liu, Fei [1 ]
Hou, Chuanqiang [1 ]
Li, Baojiang [1 ]
机构
[1] Taian Cent Hosp, Dept Breast Surg, Breast Canc Ctr, Tai An, Shandong, Peoples R China
[2] Taian Cent Hosp, Dept Cent Sterile Supply, Tai An, Shandong, Peoples R China
[3] Taian Cent Hosp, Dept Ultrason Diag, Breast Canc Ctr, Tai An, Shandong, Peoples R China
[4] Weifang Peoples Hosp, Dept Surg Oncol, Weifang, Peoples R China
来源
FRONTIERS IN ONCOLOGY | 2021年 / 11卷
关键词
triple-negative breast cancer; cancer stem cell; marker; phage; identification; PHAGE DISPLAY TECHNOLOGY; BINDING-PEPTIDE; METASTASIS;
D O I
10.3389/fonc.2021.647291
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Objective: We screened the TNBC stem cells using phage display (PD) and acquired the specific binding clones; and then the positive phage DNAs were amplified and extracted, synthesized with specific polypeptides, and labeled with fluorescein isothiocyanate (FITC). Finally, we identified the specificity of the polypeptides in vitro and in vivo. Methods: Human breast cancer cell line MDA-MB-231 and human mammary gland cell line hs578bst were chosen in our study, and MDA-MB-231 breast cancer stem cells (BCSCs) were cultured and identified by flow cytometry. The phage peptide library was screened using MDA-MB-231 BCSCs, the positive phage clones were identified by ELISA, and the DNA of the positive phages was extracted and sent to a biotechnology company for sequencing. According to the sequencing results, a specific polypeptide was synthesized and labeled with FITC. In the end, the specificity of a polypeptide to BCSCs was identified in vivo and in vitro. Results: The MDA-MB-231 BCSCs were cultured and enriched with the "serum and serum-free alternate" method. The BCSCs were found to have characteristics of CD44(+)/CD24(-/low) epithelial surface antigen (ESA) and ALDH(+) with flow cytometry. The phage was enriched to 200-fold after three rounds of screening for MDA-MB-231 BCSCs. The positive phages were sequenced; then a polypeptide named M58 was synthesized according to sequencing results. Polypeptide M58 has a specific affinity to MDA-MB-231 BCSCs in vivo and in vitro. Conclusion: Specific polypeptides binding to MDA-MB-231 BCSCs were screened out by PD screening method, which laid a theoretical foundation for the targeted therapy and further research of BCSCs.
引用
收藏
页数:9
相关论文
共 26 条
[1]   Subtyping of Triple-Negative Breast Cancer: Implications for Therapy [J].
Abramson, Vandana G. ;
Lehmann, Brian D. ;
Ballinger, Tarah J. ;
Pietenpol, Jennifer A. .
CANCER, 2015, 121 (01) :8-16
[2]   Prospective identification of tumorigenic breast cancer cells [J].
Al-Hajj, M ;
Wicha, MS ;
Benito-Hernandez, A ;
Morrison, SJ ;
Clarke, MF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (07) :3983-3988
[3]   Phage display technology: clinical applications and recent innovations [J].
Azzazy, HME ;
Highsmith, WE .
CLINICAL BIOCHEMISTRY, 2002, 35 (06) :425-445
[4]   Comprehensive Genomic Analysis Identifies Novel Subtypes and Targets of Triple-Negative Breast Cancer [J].
Burstein, Matthew D. ;
Tsimelzon, Anna ;
Poage, Graham M. ;
Coyington, Kyle R. ;
Contreras, Alejandro ;
Fuqua, Suzanne A. W. ;
Sayage, Michelle I. ;
Osborne, C. Kent ;
Hilsenbeck, Susan G. ;
Chang, Jenny C. ;
Mills, Gordon B. ;
Lau, Ching C. ;
Brown, Powel H. .
CLINICAL CANCER RESEARCH, 2015, 21 (07) :1688-1698
[5]   Prognostic Value of Cancer Stem Cells Markers in Triple-Negative Breast Cancer [J].
Collina, Francesca ;
Di Bonito, Maurizio ;
Li Bergolis, Valeria ;
De laurentiis, Michelino ;
Vitagliano, Carlo ;
Cerrone, Margherita ;
Nuzzo, Francesco ;
Cantile, Monica ;
Botti, Gerardo .
BIOMED RESEARCH INTERNATIONAL, 2015, 2015
[6]   Identification of a novel aFGF-binding peptide with anti-tumor effect on breast cancer from phage display library [J].
Dai, Xiaoyong ;
Cai, Cuizan ;
Xiao, Fei ;
Xiong, Yaoling ;
Huang, Yadong ;
Zhang, Qihao ;
Xiang, Qi ;
Lou, Guofeng ;
Lian, Mengyang ;
Su, Zhijian ;
Zheng, Qing .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2014, 445 (04) :795-801
[7]   Cancer stem cell heterogeneity: origin and new perspectives on CSC targeting [J].
Eun, Kiyoung ;
Ham, Seok Won ;
Kim, Hyunggee .
BMB REPORTS, 2017, 50 (03) :117-125
[8]   Insights into Molecular Classifications of Triple-Negative Breast Cancer: Improving Patient Selection for Treatment [J].
Garrido-Castro, Ana C. ;
Lin, Nancy U. ;
Polyak, Kornelia .
CANCER DISCOVERY, 2019, 9 (02) :176-198
[9]   Breast cancer stem cells: Multiple capacities in tumor metastasis [J].
Geng, Shao-Qing ;
Alexandrou, Aris T. ;
Li, Jian Jian .
CANCER LETTERS, 2014, 349 (01) :1-7
[10]   ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome [J].
Ginestier, Christophe ;
Hur, Min Hee ;
Charafe-Jauffret, Emmanuelle ;
Monville, Florence ;
Dutcher, Julie ;
Brown, Marty ;
Jacquemier, Jocelyne ;
Viens, Patrice ;
Kleer, Celina G. ;
Liu, Suling ;
Schott, Anne ;
Hayes, Dan ;
Birnbaum, Daniel ;
Wicha, Max S. ;
Dontu, Gabriela .
CELL STEM CELL, 2007, 1 (05) :555-567