• Synthesis of novel cell-penetrating peptide as a carrier for MCF-7 cancer cell line
  • Mahsima Heydari,1 Mohsen Farhadpour,2 Mohammad Hossein Sanati,3 Zahra Azizi,4 Amir Norouzy,5,*
    1. National Institute of Genetic Engineering and Biotechnology
    2. National Institute of Genetic Engineering and Biotechnology
    3. National Institute of Genetic Engineering and Biotechnology
    4. Tehran University of Medical Sciences
    5. National Institute of Genetic Engineering and Biotechnology


  • Introduction: In recent years, the effective delivery of pharmaceutical agents or diagnostic compounds to cells is the challenge of biology and medical science. Cell-penetrating peptides (CPPs), also known as protein transduction domains (PTDs) or membrane translocating sequence, are mostly short (5-30 amino acids) cationic peptides. CPPs have been considered as useful and promising cargo carriers due to their potential to cell entry without in a non-toxic fashion [1]. They are capable of carrying a wide range of macromolecules such as plasmids, DNA, small interfering RNAs (siRNAs), drugs (such as anticancer drugs), nanoparticles, proteins, viruses, fluorescent or radioactive compounds for Intracellular imaging [2]. The CPPs can penetrate the cells via different pathways, including energy-independent direct penetration across Biological Membranes and energy-dependent endocytosis without substantial damage to cell membrane [3]. In this study, it is reported on the development of an effective delivery; a novel cell-penetrating peptide (CPP), which is used for in vitro testing.
  • Methods: The CPP for this experiment FAM-YYYYRRRR was designed and synthesized employing solid-phase peptide synthesis method using the Fmoc strategy. 5-FAM (5-Carboxyfluorescein) serves as a fluorescent probe. It is also used p-sulfonatocalix[4]arene (CX4) with the desired peptide to enhance cell penetration and inhibit aggregation. MCF-7 cells were incubated with different concentrations of the peptide and peptide/CX4 in the range of 0.1 and 10 μM. Flow cytometry and fluorescent imaging techniques were used to evaluate the uptake of peptides into cells.
  • Results: The results showed that the FAM-YYYYRRRR peptide passing through the cell membrane in a slight matter, while the FAM-YYYYRRRR/CX4 uptake was relatively higher by the MCF7 cells in different concentrations, which shows the CX4 assisting role in the penetration.
  • Conclusion: The CX4 reduced peptide aggregation and eliminated the cytotoxicity effect. Therefore, it is suggested that the presence of the CX4 along with the peptide expedite peptide penetration into the cells in various concentrations of CPPs.
  • Keywords: MCF7, cancer cell, Cell-penetrating peptides, amphipathic peptide, p-sulfonatocalix[4]arene