[Pigment epithelium-derived factor gene therapy inhibits the growth of transplanted human hepatocellular carcinoma in nude mice].
OBJECTIVE
To evaluate the antiangiogenic property of pigment epithelium-derived factor(PEDF) in heptocarcinoma cell lines and explore its possible application in the gene therapy of human hepatocellular carcinoma (HCC).


METHODS
The gene encoding human PEDF was subcloned into lentiviral vector to generate the recombinant plasmid pLenti-PEDF. The plasmid pLenti-PEDF and two other packaging plasmids were cotransfected to 293T cells by calcium phosphate. Then HepG2 was infected with recombinant lentivirus and the expression efficiency of PEDF was analyzed by western blot. Proliferation and migration assay of human umbilical vein endothelial cells (HUVEC) was used to evaluate the biological activity of PEDF in vitro. Murine subcutaneous tumor model was established to investigate the therapeutic effects of Lenti-PEDF on HCC, and the expression of PEDF mRNA in tumor tissues was analyzed by RT-PCR.


RESULTS
Restriction enzyme digestion and DNA sequencing demonstrated that the recombinant plasmid pLenti-PEDF was constructed successfully. HepG2 secreted PEDF in the media effectively after infected with the recombinant lentivirus and this protein exhibited strong inhibitory effects on proliferation and migration of human umbilical vein endothelial cells (P less than 0.01). Intratumoral injection of Lenti-PEDF caused significant inhibition of tumor growth (P less than 0.01), and high level expression of PEDF mRNA was detected in tumor tissues by RT-PCR.


CONCLUSIONS
Our data suggest that PEDF may exert an inhibitory effect on tumor angiogenesis and PEDF gene therapy may provide a new approach for the treatment of HCC.
GAO
Yun
Y
Institute of Liver Transplantation, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
ZHANG
Wei
W
ZHAO
Liang
L
WANG
Xue-hao
XH
D000818:Animals; D049109:Cell Proliferation; D042783:Endothelial Cells / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors; D056945:Hep G2 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008113:Liver Neoplasms / Q000098:blood supply / Q000235:genetics / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic / Q000628:therapy*; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015843:Serpins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology; D000099174:Pigment Epithelium-Derived Factor
D005136:Eye Proteins; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor