Investigation into the Anticancer Activity and Apoptosis Induction of Brevinin-2R and Brevinin-2R-Conjugated PLA–PEG–PLA Nanoparticles and Strong Cell Cycle Arrest in AGS, HepG2 and KYSE-30 Cell Lines

  • Robab Hassanvand JamadiEmail author
  • Asadollah Asadi
  • Hashem Yaghoubi
  • Fariba Goudarzi


Our study aims to establish a biocompatible nanostructure for the improved delivery of anticancer peptide, Brevinin-2R, to treat human gastric adenocarcinoma (AGS), human liver hepatocellular carcinoma (HepG2) and human squamous cell carcinoma (KYSE-30) cells. Poly(l-lactide)–poly(ethylene glycol)–poly(l-lactide) (PLA–PEG–PLA) nanoparticles were synthesized, obtained by a solvent evaporation method and characterized using scanning electron microscopy (SEM), FTIR and DLS; chemically-synthesized Brevinin-2R was encapsulated in micelles. In vitro release and cell uptake assay were conducted before cytotoxicity tests. Cell cycle analysis and apoptosis study were performed through flow cytometry and Annexin-V-FlOUS cell staining. PLA–PEG–PLA nanoparticles showed a narrow-size distribution with a zeta potential of − 26.63 and a high cell internalization. Brevinin-2R-conjugated nanoparticles were spherical in shape with an increased surface charge of − 21.90. For the first time, viability tests showed that Brevinin-2R-conjugated nanoparticles were more efficient than Brevinin-2R against cancer cells causing higher rates of cell cycle arrest and apoptosis induction. Our new findings demonstrate the potential of PLA–PEG–PLA nanoparticles to boost the anticancer effect and improve the delivery of Brevinin-2R. The study of Brevinin-2R-loaded nanoparticles indicated noticeable results in terms of novel cancer therapy. PLA–PEG–PLA nanoparticles can act as a biocompatible delivery platform to take the advantage of Brevinin-2R toward cancer cells. This is a novel study as the Brevinin-2R-conjugated nanoparticles and applied approaches have not been already reported.


Apoptosis induction Brevinin-2R-conjugated nanoparticle Cell cycle arrest Cell internalization Drug loading Enhanced anticancer efficiency 



This research was financially supported by the University of Mohaghegh Ardabili, Ardabil, Iran. The authors would like to acknowledge Jennifer Mason for proofreading the article. We are also grateful to the contribution of Dr. Hossein Akbari from Ardabil Azad University and Mehdi Hassanvand Jamadi from University of Shahid Beheshti to statistical analysis and critical reviewing of manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research Involving Human and Animal Participants

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Informed consent was obtained from all individual participants included in the study.


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Biology, Faculty of SciencesUniversity of Mohaghegh ArdabiliArdabilIran
  2. 2.Department of BiologyArdabil Branch, Islamic Azad UniversityArdabilIran

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