Enhancing anticancer effect of gefitinib across the blood-brain barrier model using liposomes modified with one ?-helical cell-penetrating peptide or glutathione and Tween 80
Journal
International Journal of Molecular Sciences
Journal Volume
17
Journal Issue
12
Date Issued
2016
Author(s)
Abstract
Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIa), such as gefitinib, have been demonstrated to effectively treat the patients of extracranial non-small cell lung cancer (NSCLC). However, these patients often develop brain metastasis (BM) during their disease course. The major obstacle to treat BM is the limited penetration of anticancer drugs across the blood-brain barrier (BBB). In the present study, we utilized gefitinib-loaded liposomes with different modifications to improve gefitinib delivery across the in vitro BBB model of bEnd.3 cells. Gefitinib was encapsulated in small unilamellar liposomes modified with glutathione (GSH) and Tween 80 (SUV-G+T; one ligand plus one surfactant) or RF (SUV-RF; one α-helical cell-penetrating peptide). GSH, Tween 80, and RF were tested by the sulforhodamine B (SRB) assay to find their non-cytotoxic concentrations on bEnd.3 cells. The enhancement on gefitinib across the BBB was evaluated by cytotoxicity assay on human lung adenocarcinoma PC9 cells under the bEnd.3 cells grown on the transwell inserts. Our findings showed that gefitinib incorporated in SUV-G+T or SUV-RF across the bEnd.3 cells significantly reduced the viability of PC9 cells more than that of free gefitinib. Furthermore, SUV-RF showed no cytotoxicity on bEnd.3 cells and did not affect the transendothelial electrical resistance (TEER) and transendothelial permeability of sodium fluorescein across the BBB model. Moreover, flow cytometry and confocal laser scanning microscopy were employed to evaluate the endocytosis pathways of SUV-RF. The results indicated that the uptake into bEnd.3 cells was mainly through adsorptive-mediated mechanism via electrostatic interaction and partially through clathrin-mediated endocytosis. In conclusion, cell penetrating peptide-conjugated SUV-RF shed light on improving drug transport across the BBB via modulating the transcytosis pathway(s). ? 2016 by the authors; licensee MDPI, Basel, Switzerland.
Subjects
Blood-brain barrier (BBB); Gefitinib; Liposomes; Lung cancer; Peptides
SDGs
Other Subjects
cell penetrating peptide; gefitinib; glutathione; liposome; polysorbate 80; cell penetrating peptide; gefitinib; polysorbate; quinazoline derivative; animal cell; antineoplastic activity; Article; blood brain barrier; cell viability; confocal laser scanning microscopy; controlled study; cytotoxicity assay; drug delivery system; drug uptake; endocytosis; flow cytometry; IC50; lung adenocarcinoma; membrane permeability; nanoencapsulation; nonhuman; particle size; transendothelial permeability; transepithelial resistance; transmission electron microscopy; zeta potential; blood brain barrier; cell survival; chemistry; confocal microscopy; drug effects; human; lung tumor; metabolism; tumor cell line; Blood-Brain Barrier; Cell Line, Tumor; Cell Survival; Cell-Penetrating Peptides; Glutathione; Humans; Liposomes; Lung Neoplasms; Microscopy, Confocal; Microscopy, Electron, Transmission; Polysorbates; Quinazolines
Publisher
MDPI AG
Type
journal article