Metastatic prostate cancer cell-specific phage-like particles as a targeted gene-delivery system.
BACKGROUND
One of the cardinal requirements for effective therapeutic management of tumors is the selective delivery of cancer drugs to the right site by ligand-decorated nanomedicines. Screening of 2 × 109 clone landscape phage library provides a reliable avenue for generating protein ligands specific for tumor cells. It was shown that selective phage proteins derived from landscape phage libraries against breast and prostate cancer cells are able to navigate drug or siRNA loaded liposomes to corresponding cancer cells with minimal toxicity to non-neoplastic cells. In an alternative platform, glioma cell-specific phage proteins were used for assembling in vivo cancer-specific phage-like particles, named 'phagemid infective particles' as targeted gene-delivery vehicles.


METHODS
To extend the panel of anticancer cell phages, we have screened a 2 × 109 clone landscape phage library f8/8 to select phage clones specific for metastatic prostate cancer cell PC-3M. The phage clones were characterized for their selective interaction with PC-3M cells using phage capture assay, immunofluorescence microscopy and electron microscopy. A prostate cancer selective phage was converted to phage-like particles harboring emerald green fluorescent protein.


RESULTS
Phage clone EPTHSWAT (designated by the sequence of inserted peptide) was found to be most selective for PC-3M cells and was observed to internalize PC-3M cells as revealed by immunofluorescence microscopy and electron microscopy. Conversion of this phage to phage-like particles harboring emerald green fluorescent protein and the expression of emerald green fluorescent protein in the phage-like particles treated PC-3M cells showed potential of adoption of this phage-like particle in prostate cancer therapeutic gene delivery.


CONCLUSIONS
Successful employment of phage-like particles expressing emerald green fluorescent protein genes targeted to prostate cancer cells PC-3M confirms a prospect of their use for targeted delivery of therapeutic genes to cancer cells.
Cancer Research Center, Columbia, MO 65201, USA. EisenstarkA@missouri.edu.
Olusegun A
OA
Fagbohun
Robert A
RA
Kazmierczak
Valery A
VA
Petrenko
Abraham
A
Eisenstark
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics*; D045744:Cell Line, Tumor; D004705:Endocytosis; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D058990:Molecular Targeted Therapy / Q000379:methods; D009362:Neoplasm Metastasis; D019151:Peptide Library; D011471:Prostatic Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy; D014771:Virion / Q000235:genetics*
D019151:Peptide Library; D049452:Green Fluorescent Proteins