The Enhanced Cytotoxic Effects of the p28-Apoptin Chimeric Protein As A Novel Anti-Cancer Agent on Breast Cancer Cell Lines.
Journal
Drug research
ISSN: 2194-9387
Titre abrégé: Drug Res (Stuttg)
Pays: Germany
ID NLM: 101602406
Informations de publication
Date de publication:
Feb 2019
Feb 2019
Historique:
pubmed:
31
7
2018
medline:
10
7
2019
entrez:
31
7
2018
Statut:
ppublish
Résumé
Peptide-based drugs have shown promising results in overcoming the limitations of chemotherapeutic drugs by providing a targeted therapy approach to cancer. However, the response rate of targeted therapies is limited, in large part due to the intra- and inter-heterogeneity of tumors. In this study, we engineered a novel chimeric protein composed of the p28 peptide as a tumor-homing killer peptide and apoptin as a killer peptide. We evaluated its cytotoxicity against MCF7 and MDA-MB-231 breast cancer cells and HEK-293 normal cells by the MTT assay. Different linkers were evaluated when designing the chimeric protein. Three-dimensional structure predictions of chimeric proteins with different linkers were carried out by Modeller 9.19, and their validation and analysis were performed by RAMPAGE. Results showed that a cleavable linker, including furin cleavage sites, is preferred over other linkers. The chimeric protein was then successfully expressed in E. coli and purified by affinity chromatography under native conditions, then confirmed by SDS-PAGE and Western blot analysis. Compared with apoptin alone, the chimeric protein showed significantly higher toxicity against breast cancer cell lines in a dose-dependent manner. The IC This study demonstrates that fusion of p28 peptide to a potent protein could provide an effective method for tumor targeting. Further, in vitro and in vivo studies of this novel chimeric protein are underway.
Sections du résumé
BACKGROUNDS
BACKGROUND
Peptide-based drugs have shown promising results in overcoming the limitations of chemotherapeutic drugs by providing a targeted therapy approach to cancer. However, the response rate of targeted therapies is limited, in large part due to the intra- and inter-heterogeneity of tumors.
METHODS
METHODS
In this study, we engineered a novel chimeric protein composed of the p28 peptide as a tumor-homing killer peptide and apoptin as a killer peptide. We evaluated its cytotoxicity against MCF7 and MDA-MB-231 breast cancer cells and HEK-293 normal cells by the MTT assay. Different linkers were evaluated when designing the chimeric protein. Three-dimensional structure predictions of chimeric proteins with different linkers were carried out by Modeller 9.19, and their validation and analysis were performed by RAMPAGE.
RESULTS
RESULTS
Results showed that a cleavable linker, including furin cleavage sites, is preferred over other linkers. The chimeric protein was then successfully expressed in E. coli and purified by affinity chromatography under native conditions, then confirmed by SDS-PAGE and Western blot analysis. Compared with apoptin alone, the chimeric protein showed significantly higher toxicity against breast cancer cell lines in a dose-dependent manner. The IC
CONCLUSION
CONCLUSIONS
This study demonstrates that fusion of p28 peptide to a potent protein could provide an effective method for tumor targeting. Further, in vitro and in vivo studies of this novel chimeric protein are underway.
Substances chimiques
Antineoplastic Agents
0
Recombinant Fusion Proteins
0
p28-apoptin protein, human
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
144-150Informations de copyright
© Georg Thieme Verlag KG Stuttgart · New York.
Déclaration de conflit d'intérêts
The authors declare that there is no conflict of interest regarding the publication of this article.