Nano-encapsulation of a novel anti-Ran-GTPase peptide for blockade of regulator of chromosome condensation 1 (RCC1) function in MDA-MB-231 breast cancer cells.
Ran is a small ras-related GTPase and is highly expressed in aggressive breast carcinoma. Overexpression induces malignant transformation and drives metastatic growth. We have designed a novel series of anti-Ran-GTPase peptides, which prevents Ran hydrolysis and activation, and although they display effectiveness in silico, peptide activity is suboptimal in vitro due to reduced bioavailability and poor delivery. To overcome this drawback, we delivered an anti-Ran-GTPase peptide using encapsulation in PLGA-based nanoparticles (NP). Formulation variables within a double emulsion solvent evaporation technique were controlled to optimise physicochemical properties. NP were spherical and negatively charged with a mean diameter of 182-277nm. Peptide integrity and stability were maintained after encapsulation and release kinetics followed a sustained profile. We were interested in the relationship between cellular uptake and poly(ethylene glycol) (PEG) in the NP matrix, with results showing enhanced in vitro uptake with increasing PEG content. Peptide-loaded, pegylated (10% PEG)-PLGA NP induced significant cytotoxic and apoptotic effects in MDA-MB-231 breast cancer cells, with no evidence of similar effects in cells pulsed with free peptide. Western blot analysis showed that encapsulated peptide interfered with the proposed signal transduction pathway of the Ran gene. Our novel blockade peptide prevented Ran activation by blockage of regulator of chromosome condensation 1 (RCC1) following peptide release directly in the cytoplasm once endocytosis of the peptide-loaded nanoparticle has occurred. RCC1 blockage was effective only when a nanoparticulate delivery approach was adopted.
School of Pharmacy and Pharmaceutical Sciences, Saad Centre for Pharmacy and Diabetes, Ulster University, Cromore Road, Coleraine, Co. Londonderry, BT52 1SA, UK; Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Tanta, Tanta, Egypt.
Yusuf A
YA
Haggag
Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK.
Kyle B
KB
Matchett
Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK; Institute of Cancer Therapeutics, University of Bradford, Bradford, UK.
El-Habib
el-H
Dakir
Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK.
Paul
P
Buchanan
Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Tanta, Tanta, Egypt.
Mohammed A
MA
Osman
Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Tanta, Tanta, Egypt.
Sanaa A
SA
Elgizawy
Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK; Institute of Cancer Therapeutics, University of Bradford, Bradford, UK; IDT (Imhotep Diagnostics and Therapeutics), Europa Tool House, Springbank, Industrial Estate, Dunmurry, Northern Ireland, UK.
Mohamed
M
El-Tanani
Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Tanta, Tanta, Egypt; Sunderland Pharmacy School, Department of Pharmacy, Health and Well Being, University of Sunderland, Sunderland SR1 3SD, UK.
Ahmed M
AM
Faheem
School of Pharmacy and Pharmaceutical Sciences, Saad Centre for Pharmacy and Diabetes, Ulster University, Cromore Road, Coleraine, Co. Londonderry, BT52 1SA, UK. Electronic address: p.mccarron@ulster.ac.uk.
Paul A
PA
McCarron
D001943:Breast Neoplasms / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000502:physiology; D045744:Cell Line, Tumor; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry; D016503:Drug Delivery Systems / Q000379:methods; D005260:Female; D020690:GTPase-Activating Proteins / Q000008:administration & dosage / Q000737:chemistry*; D020662:Guanine Nucleotide Exchange Factors / Q000037:antagonists & inhibitors* / Q000502:physiology; D006801:Humans; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000502:physiology; D011091:Polyesters / Q000008:administration & dosage / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000008:administration & dosage / Q000737:chemistry*
D018797:Cell Cycle Proteins; D004337:Drug Carriers; D020690:GTPase-Activating Proteins; D020662:Guanine Nucleotide Exchange Factors; D009687:Nuclear Proteins; D011091:Polyesters; C092317:RANGAP1 protein, human; C062521:RCC1 protein, human; C000589473:polyethylene glycol-poly(lactide-co-glycolide); D011092:Polyethylene Glycols
Anti-Ran-GTPase peptide; Breast cancer; Double emulsion; Drug delivery; Nanoparticle; PLGA