Screening and identification of novel specific markers of breast cancer stem cells

Liu,Tingting; Li,Baojiang; Jiang,Yunyun; Zheng,Chunhui; Zhang,Li; Wang,Yongsheng

doi:10.3892/ol.2019.10535

Screening and identification of novel specific markers of breast cancer stem cells

Authors:
- Tingting Liu
- Baojiang Li
- Yunyun Jiang
- Chunhui Zheng
- Li Zhang
- Yongsheng Wang
View Affiliations

Affiliations: Breast Cancer Center, Shandong Cancer Hospital and Institute Affiliated to Shandong University, Jinan, Shandong 250117, P.R. China, Department of Breast Surgery, Tai'an Central Hospital, Tai'an, Shandong 271000, P.R. China, Department of Rehabilitation Medicine, Tai'an Central Hospital, Tai'an, Shandong 271000, P.R. China, Department of Oncology Surgery, Weifang People's Hospital, Weifang, Shandong 261000, P.R. China, Department of Breast Oncology, Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University Cancer Institute and Hospital/Ministry of Education, Tianjin 300060, P.R. China
Published online on: June 27, 2019 https://doi.org/10.3892/ol.2019.10535
Pages: 2262-2269
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Breast cancer is the leading cause of death among women worldwide. Until recent years, triple negative breast cancer could be divided into 6 types according to different biomarkers with the development of sequence and microarray technology. However, these results rarely have therapeutic impact and still lack validation with the string criteria of clinical studies. Therefore, the present study aimed to screen novel markers of breast cancer stem cells and to verify the specificity in vitro and in vivo. In the present study, screening for phages specifically binding to breast cancer stem cells was performed, positive phage DNAs were extracted, and polypeptides were synthesized and labeled with FITC. The specificity of the polypeptides was identified in vitro and in vivo. Breast cancer stem cells were cultured and identified by flow cytometry. A phage random‑peptide library was amplified and screened by culturing with breast cancer cells and breast cancer stem cells. The positive phage was identified by ELISA, and positive phage DNA was extracted. The DNA pellet was isolated and sent for external sequencing with the primer ‑96 gIII. Based on the sequencing results, a polypeptide was synthesized and labeled with FITC. The specificity to breast cancer stem cells was identified in vivo and vitro. Following three rounds of screening, the phage was enriched ~200‑fold. Immunofluorescence demonstrated that two randomly selected phage clones, B8 and A3, had specific affinity to breast cancer stem cells. The results of the present study indicated that phage polypeptides that specifically bind to breast cancer stem cells were successfully screened through stem cell enrichment and phage display technology, which may be beneficial for targeted therapy and further study of breast cancer stem cells.

View Figures

View References

1	Feuer EJ, Wun LM, Boring CC, Flanders WD, Timmel MJ and Tong T: The lifetime risk of developing breast cancer. J Natl Cancer Inst. 85:892–897. 1993. View Article : Google Scholar : PubMed/NCBI
2	So WK, Choi KC, Chan CW and Chair SY: Age-related differences in the quality of life of Chinese women undergoing adjuvant therapy for breast cancer. Res Gerontol Nurs. 29–26. 2011.
3	Zhu D, Lam DH, Purwanti YI, Goh SL, Wu C, Zeng J, Fan W and Wang S: Systemic delivery of fusogenic membrane glycoprotein-expressing neural stem cells to selectively kill tumor cells. Mol Ther. 21:1621–1630. 2013. View Article : Google Scholar : PubMed/NCBI
4	Luo M, Clouthier SG, Deol Y, Liu S, Nagrath S, Azizi E and Wicha MS: Breast cancer stem cells: Current advances and clinical implications. Methods Mol Biol. 1293:1–49. 2015. View Article : Google Scholar : PubMed/NCBI
5	Hyvönen M and Laakkonen P: Identification and characterization of homing peptides using in vivo peptide phage display. Methods Mol Biol. 1324:205–222. 2015. View Article : Google Scholar : PubMed/NCBI
6	Tiede C, Tang AA, Deacon SE, Mandal U, Nettleship JE, Owen RL, George SE, Harrison DJ, Owens RJ, Tomlinson DC and McPherson MJ: Adhiron: A stable and versatile peptide display scaffold for molecular recognition applications. Protein Eng Des Sel. 27:145–55. 2014. View Article : Google Scholar : PubMed/NCBI
7	Ajani JA, Song S, Hochster HS and Steinberg IB: Cancer stem cells: The promise and the potential. Semin Oncol. 42 (Suppl 1):S3–S17. 2015. View Article : Google Scholar : PubMed/NCBI
8	Li X and Mao C: Using phage as a platform to select cancer cell-targeting peptides. Methods Mol Biol. 1108:57–68. 2014. View Article : Google Scholar : PubMed/NCBI
9	Wang T, Hartner WC, Gillespie JW, Praveen KP, Yang S, Mei LA, Petrenko VA and Torchilin VP: Enhanced tumor delivery and antitumor activity in vivo of liposomal doxorubicin modified with MCF-7-specific phage fusion protein. Nanomedicine. 10:421–430. 2014. View Article : Google Scholar : PubMed/NCBI
10	Pouyanfard S, Bamdad T, Hashemi H, Bandehpour M and Kazemi B: Induction of protective anti-CTL epitope responses against HER-2-positive breast cancer based on multivalent T7 phage nanoparticles. PLoS One. 7:e495392012. View Article : Google Scholar : PubMed/NCBI
11	Finlay-Schultz J, Cittelly DM, Hendricks P, Patel P, Kabos P, Jacobsen BM, Richer JK and Sartorius CA: Progesterone downregulation of miR-141 contributes to expansion of stem-like breast cancer cells through maintenance of progesterone receptor and Stat5a. Oncogene. 34:3676–3687. 2015. View Article : Google Scholar : PubMed/NCBI
12	Lv YG, Wang T, Yuan SF, Li NL, Chen JH, Zhao AZ, Ling R and Wang L: T-vector and in vivo recombination as tools to construct a large antibody library of breast cancer. Hybridoma (Larchmt). 29:251–254. 2010. View Article : Google Scholar : PubMed/NCBI
13	Petrenko VA and Jayanna PK: Phage protein-targeted cancer nanomedicines. FEBS Lett. 588:341–349. 2014. View Article : Google Scholar : PubMed/NCBI
14	Park HY, Lee KJ, Lee SJ and Yoon MY: Screening of peptides bound to breast cancer stem cell specific surface marker CD44 by phage display. Mol Biotechnol. 51:212–220. 2012. View Article : Google Scholar : PubMed/NCBI
15	Nemudraya AA, Kuligina EV, Ilyichev AA, Fomin AS, Stepanov GA, Savelyeva AV, Koval OA and Richter VA: Selection of antitumor displayed peptides for the specific delivery of the anticancer drug lactaptin. Oncol Lett. 12:4547–4555. 2016. View Article : Google Scholar : PubMed/NCBI
16	Yang H, Brand JS, Fang F, Chiesa F, Johansson AL, Hall P and Czene K: Time-dependent risk of depression, anxiety, and stress-related disorders in patients with invasive and in situ breast cancer. Int J Cancer. 140:841–852. 2017. View Article : Google Scholar : PubMed/NCBI
17	Yang F, Xu J, Tang L and Guan X: Breast cancer stem cell: The roles and therapeutic implications. Cell Mol Life Sci. 74:951–966. 2017. View Article : Google Scholar : PubMed/NCBI
18	Asad AS, Moreno Ayala MA, Gottardo MF, Zuccato C, Nicola Candia AJ, Zanetti FA, Seilicovich A and Candolfi M: Viral gene therapy for breast cancer: Progress and challenges. Exp Opin Biol Ther. 17:945–959. 2017. View Article : Google Scholar
19	Alferez DG, Simões BM, Howell SJ and Clarke RB: The role of steroid hormones in breast and effects on cancer stem cells. Curr Stem Cell Rep. 4:81–94. 2018. View Article : Google Scholar : PubMed/NCBI
20	Wolfe AR and Woodward WA: Breast cancer stem cell correlates as Predicative factors for radiation therapy. Semin Radiat Oncol. 25:251–259. 2015. View Article : Google Scholar : PubMed/NCBI
21	To K, Fotovati A, Reipas KM, Law JH, Hu K, Wang J, Astanehe A, Davies AH, Lee L, Stratford AL, et al: Y-box binding protein-1 induces the expression of CD44 and CD49f leading to enhanced self-renewal, mammo-sphere growth, and drug resistance. Cancer Res. 70:2840–2851. 2010. View Article : Google Scholar : PubMed/NCBI
22	Liu F, Qi CL, Kong M, Liu TT, Li L and Li BJ: Screening specific polypeptides of breast cancer stem cells from a phage display random peptide library. Oncol Lett. 12:4727–4731. 2016. View Article : Google Scholar : PubMed/NCBI