Silencing of the metastasis-linked gene, AEG-1, using siRNA-loaded cholamine surface-modified gelatin nanoparticles in the breast carcinoma cell line MCF-7.
Cholamine surface-modified gelatin nanoparticles prepared by the double desolvation method using acetone as a dehydrating agent were selected and potentially evaluated as non viral vectors of siRNA targeting a metastatic gene AEG-1 in MCF-7 breast carcinoma cells. The ability of modified gelatin nanoparticle to complex and deliver siRNA for gene silencing was investigated. Hence, Particle size, surface charge (zeta potential) and morphology of siRNA/Gelatin nanoparticles (siGNPs) were characterized via dynamic light scattering (DLS), scanning electron microscopy (SEM) and transmission electron microscope (TEM). Moreover, the nanoparticles cytotoxicity, loading efficiency and interaction with MCF-7 human breast carcinoma cells were evaluated. Cationized GNPs of mean size range of 174nm and PDI of 0.101 were produced. The loading efficiency of siGNPs at a Nitrogen/Phosphate (N/P) ratio (w/w) of 200:1 was approximately 96%. Cellular uptake was evaluated after FITC conjugation where the particles produced high transfection efficiency. Finally, ELISA analysis of AEG-1/MTDH expression demonstrated the gene silencing effect of siGNPs, as more than 75% MTDH protein were inhibited. Our data indicate that cholamine modified GNPs pose a promising non-viral siRNA carrier for altering gene expression in MCF-7 breast cancer cells with many advantages such as relatively high gene transfection efficiency and efficient silencing ability.
Department of Pharmaceutical Biology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, Egypt.
Salma M
SM
Abozeid
Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt; Department of Pharmaceutical Technology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, Egypt. Electronic address: r_hathout@yahoo.com.
Rania M
RM
Hathout
Department of Pharmaceutical Biology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, Egypt.
Khaled
K
Abou-Aisha
D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D002412:Cations; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D004705:Endocytosis / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005453:Fluorescence; D005780:Gelatin / Q000737:chemistry*; D020868:Gene Silencing / Q000187:drug effects; D005976:Glutaral / Q000737:chemistry; D006801:Humans; D061986:MCF-7 Cells; D008565:Membrane Proteins; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009362:Neoplasm Metastasis; D010316:Particle Size; D034741:RNA, Small Interfering / Q000378:metabolism*; D016601:RNA-Binding Proteins; D034421:Sus scrofa; D014162:Transfection; D050337:Trimethyl Ammonium Compounds / Q000494:pharmacology*
D002412:Cations; D015815:Cell Adhesion Molecules; C483039:MTDH protein, human; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D050337:Trimethyl Ammonium Compounds; C522723:cholamine; D005780:Gelatin; D016650:Fluorescein-5-isothiocyanate; D005976:Glutaral
AEG-1; Breast cancer; Cholamine; Gelatin; Gene delivery; MTDH; Metastasis; Nanoparticles; Silencing