Screening specific polypeptides of breast cancer stem cells from a phage display random peptide library

Liu,Fei; Qi,Chun‑Ling; Kong,Mian; Liu,Ting‑Ting; Li,Lei; Li,Bao‑Jiang

doi:10.3892/ol.2016.5248

Screening specific polypeptides of breast cancer stem cells from a phage display random peptide library

Authors:
- Fei Liu
- Chun‑Ling Qi
- Mian Kong
- Ting‑Ting Liu
- Lei Li
- Bao‑Jiang Li
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Affiliations: Cancer Center, Taian Hospital, Taishan Medical College, Taian, Shandong 271000, P.R. China, Clinical Test Center, Taian Hospital, Taishan Medical College, Taian, Shandong 271000, P.R. China, Department of Thyroid Breast Surgery, The First People's Hospital of Jining, Jining, Shandong 272000, P.R. China
Published online on: October 13, 2016 https://doi.org/10.3892/ol.2016.5248
Pages: 4727-4731

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Abstract

The present study aimed to identify polypeptides that specifically bond to breast cancer stem cells from a phage display random 12 peptide library, in addition to the affinity and specificity of polypeptides. A phage display random 12 peptide library was screened using breast cancer stem cells as targets isolated from the MDA‑MB‑231 cell line using the serum‑free culture technique with hs578bst and MDA‑MB‑231 cells as subtract‑screening cells. Positive and specific binding clones were amplified and sent for sequencing. The affinity and specificity of the positive clones were subsequently identified by ELISA and 3,3'-diaminobenzidine staining. The results demonstrated that phages were gathered ~500 times following three rounds of biopanning. ELISA identified that the affinity to breast cancer stem cells of the no. 6 phage was 6.14 times higher than that in the control group. In addition, immunohistochemistry observed that the no. 6 phage exhibited high-specificity bonding to breast cancer stem cells, and the peptide sequence of the positive phage was GYSASRSTIPGK following DNA sequencing and translation. Thus, the present study isolated a specific peptide that bonds to breast cancer stem cells from a phage display random peptide library, which may facilitate further studies regarding the stem cell‑targeted therapy of breast cancer.

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