IGF-I triple helix strategy in hepatoma treatment.
OBJECTIVE
To investigate the effect of gene therapy for hepatocellular carcinoma based on inhibition of cellular IGF-I expression, the technique of IGF-I triple helix was investigated in mice developing programmed hepatoma.


METHODS
mhAT1F1 mouse hepatoma cell line was transfected in vitro with IGF-I triple helix expression vector (pMT-AG-TH) or with IGF-I antisense expression vector (pMT-Anti-IGF-I). 10 x 10(6) transfected cells of either triple helix or antisense type were inoculated intraperitonealy into transgenic ATIIITB6 mice developing genetically programmed hepatoma (mice die between the age of 6 and 7 months). In parallel, human cell cultures established from surgically removed hepatomas were investigated.


RESULTS
mhAT1F1 and human primary cell cultures, transfected with pMT-AG-TH or pMT-Anti-IGF-I vectors resulted in total inhibition of IGF-I demonstrated by immunocytochemical and Northern blot techniques. Transfected cells changed their phenotype and recovered major histocompatibility complex I expression showed by fluorescence-activated cell sorting analysis and Western blot. Moreover, two phenomena were observed in IGF-I "antisense" or "triple helix" transfected cells: 1) the apoptosis, demonstrated by TUNEL technique; 2) the presence of IL-6 simultaneously with disappearance of tumor necrosis factor-alpha and IL-10, investigated by reverse transcriptase-polymerase chain reaction technique. In in vivo experiments, injection of murine transfected cells into mice in terminal-phase prolonged their survival 3-4 months in 100% of cases, as well in "antisense" group (8/8) as in "triple helix" group (10/10).


CONCLUSIONS
Injection of hepatoma cells transfected with IGF-I triple helix expression vector, and showing immunogenic and apoptotic characteristics, can constitute an effective cellular therapy against hepatocellular carcinoma.
Upegui-Gonzalez
L C
LC
Lab. INSERM, Université Paris VII, Hôpital Bichat, Paris, France.
Ly
A
A
Sierzega
M
M
Jarocki
P
P
Trojan
L
L
Duc
H T
HT
Pan
Y
Y
Shevelev
A
A
Henin
D
D
Anthony
D
D
Nowak
W
W
Popiela
T
T
Trojan
J
J
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000037:antagonists & inhibitors* / Q000235:genetics; D015850:Interleukin-6 / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008822:Mice, Transgenic; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000473:pathology
D015850:Interleukin-6; D007334:Insulin-Like Growth Factor I