A study on screening and antitumor effect of CD55-specific ligand peptide in cervical cancer cells
Authors Liu G, Yin Q, Ji H, Wang Y, Liu H, Jiang L, Zhu F, Li B
Received 2 August 2018
Accepted for publication 10 October 2018
Published 13 November 2018 Volume 2018:12 Pages 3899—3912
Checked for plagiarism Yes
Review by Single-blind
Peer reviewer comments 2
Editor who approved publication: Dr Anastasios Lymperopoulos
Guoxiang Liu, Qifeng Yin, Huanhuan Ji, Yujuan Wang, Huihui Liu, Liangqian Jiang, Feng Zhu, Bing Li
Department of Genetics and Cell Biology, Basic Medical College, Qingdao University, Qingdao 266071, People’s Republic of China
Background: To improve the targeting ability of antitumor drugs, we identified the antigens with high expression on the surface of tumor cells associated with tumor escape, such as the complement regulatory protein CD55 molecule, which is also known as the decay accelerating factor. In this study, phage display technology was used to screen and identify CD55-specific ligand peptide (CD55sp) bound to CD55 molecule on the surface of cervical cancer HeLa cells. We then explored the role of this peptide in inhibiting the growth of cervical cancer cells in vitro. Our characterization of CD55sp will provide implication for tumor target therapy.
Methods: The phage bound to the surface of HeLa cells were isolated by phage display technology. Positive phage clones were identified by ELISA. Phage was then amplified and determined by agarose gel electrophoresis after monoclonal DNA extraction. DNA sequencing and bioinformatical analysis were conducted to obtain specific ligand peptides. Flow cytometry and immunofluorescence were used to measure the expression of CD55 molecule on the surface of tumor and normal cells. Subsequently, the effects of CD55sp on the proliferation and apoptosis of HeLa and SiHa cells were determined by Cell Counting Kit-8 (CCK-8), flow cytometry, and TUNEL assay, respectively. The morphology of apoptotic cells was examined by electron microscope. The distribution of Cleaved caspase-3 was detected by immunofluorescence. The expression of bcl-2 and Cleaved caspase-3 were determined by Western blot.
Results: The results showed that the peptide (QVNGLGERSQQM) can bind to the CD55 molecule on the surface of cervical cancer HeLa and SiHa cells as a ligand peptide. It can also effectively inhibit the proliferation of cervical cancer cells and induce cell apoptosis.
Conclusion: This study demonstrates that CD55sp screened by phage display technology plays a strong antitumor role.
Keywords: phage display, cervical cancer, CD55sp, ligand peptide, apoptosis
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