Gene-carried hepatoma targeting complex induced high gene transfection efficiency with low toxicity and significant antitumor activity.
BACKGROUND
The success of gene transfection is largely dependent on the development of a vehicle or vector that can efficiently deliver a gene to cells with minimal toxicity.


METHODS
A liver cancer-targeted specific peptide (FQHPSF sequence) was successfully synthesized and linked with chitosan-linked polyethylenimine (CP) to form a new targeted gene delivery vector called CPT (CP/peptide). The structure of CPT was confirmed by (1)H nuclear magnetic resonance spectroscopy and ultraviolet spectrophotometry. The particle size of CPT/ DNA complexes was measured using laser diffraction spectrometry and the cytotoxicity of the copolymer was evaluated by methylthiazol tetrazolium method. The transfection efficiency evaluation of the CP copolymer was performed using luciferase activity assay. Cellular internalization of the CP/DNA complex was observed under confocal laser scanning microscopy. The targeting specificity of the polymer coupled to peptide was measured by competitive inhibition transfection study. The liver targeting specificity of the CPT copolymer in vivo was demonstrated by combining the copolymer with a therapeutic gene, interleukin-12, and assessed by its abilities in suppressing the growth of ascites tumor in mouse model.


RESULTS
The results showed that the liver cancer-targeted specific peptide was successfully synthesized and linked with CP to form a new targeted gene delivery vector called CPT. The composition of CPT was confirmed and the vector showed low cytotoxicity and strong targeting specificity to liver tumors in vitro. The in vivo study results showed that interleukin-12 delivered by the new gene vector CPT/DNA significantly enhanced the antitumor effect on ascites tumor-bearing imprinting control region mice as compared with polyethylenimine (25 kDa), CP, and other controls, which further demonstrate the targeting specificity of the new synthesized polymer.


CONCLUSIONS
The synthesized CPT copolymer was proven to be an effective liver cancer-targeted vector for therapeutic gene delivery, which could be a potential candidate for targeted cancer gene therapy.
Institute of Pharmaceutics, Zhejiang University, Hangzhou, China.
Qing-Qing
QQ
Zhao
Yu-Lan
YL
Hu
Yang
Y
Zhou
Ni
N
Li
Min
M
Han
Gu-Ping
GP
Tang
Feng
F
Qiu
Yasuhiko
Y
Tabata
Jian-Qing
JQ
Gao
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D048271:Chitosan / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D006801:Humans; D018664:Interleukin-12 / Q000235:genetics; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008813:Mice, Inbred ICR; D009842:Oligopeptides / Q000737:chemistry; D010316:Particle Size; D011094:Polyethyleneimine / Q000737:chemistry; D016019:Survival Analysis; D014162:Transfection / Q000379:methods*; D023041:Xenograft Model Antitumor Assays
D009842:Oligopeptides; D018664:Interleukin-12; D011094:Polyethyleneimine; D004247:DNA; D048271:Chitosan; D016650:Fluorescein-5-isothiocyanate