Premium
Application of a platform nanotechnology for anti‐NF‐kB siRNA delivery to metastatic melanoma
Author(s) -
Pan Hua,
Hou Kirk,
Allen Stacy,
Scott Mike,
Wickline Samuel
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.620.8
Subject(s) - in vivo , gene knockdown , small interfering rna , melanoma , in vitro , flow cytometry , chemistry , cancer research , metastasis , microbiology and biotechnology , medicine , cancer , biology , transfection , apoptosis , biochemistry , gene
A cationic peptide‐based nanotechnology system for siRNA delivery recently developed in our lab not only overcomes limitations of serum instability, endosomal entrapment and vascular constraint at the disease sites of interest, but also exhibits an exceptional safety profile (Hou et al, ACS Nano and Biomaterials; 2014). The objective of this study was to address the role of NF‐kB in melanoma by inhibiting the canonical pathway component, p65, with 55nm diameter nanoparticles carrying anti‐p65 siRNA (p65 NP) in mice with pulmonary metastatic melanoma. Flow cytometry and fluorescence microscopy were used to confirm cytoplasmic delivery of siRNA to B16F10 cells in vitro. RT2‐PCR and western blot were used to confirm p65 knockdown at both mRNA and protein levels after the anti‐p65 siRNA NP treatment. For in vivo studies, C57BL/6J mice received injections of 1/2 million B16F10 cells to create lung metastasis. Three consecutive i.v. treatments of siRNA NP were administered on day 4, 5, 6 after B16F10 cell injections, and mice were sacrificed on day 7 for metastasis evaluation. Our results indicate that siRNA NP are readily delivered into the cytoplasm of B16F10 cells and into lungs parenchyma. Both p65 mRNA (88% less) and protein were substantially depleted in B16F10 cells (n=6).After treatment, melanoma lung metastasis (black spots in lung) were dramatically reduced as compared with controls (n=10 per group).In conclusion, this platform technology enables effective cellular delivery of specific siRNAs against p65 both in vitro and in vivo, which appears effective in altering the natural history of melanoma lung metastasis within 7 days in this model.