[The effect of antisense H-ras on growth and metastasis of a high metastatic tumor model of human hepatoma in nude mice LCI-D20].
OBJECTIVE
To investigate if treatment with antisense H-ras oligodeoxynucleotides (ODNs) modulates tumor growth, apoptosis and metastasis of a high metastatic tumor model of human hepatocellular carcinoma (HCC) in nude mice LCI-D20, in which over-expression of H-ras has been identified.


METHODS
LCI-D20 cells in primary culture were treated with 10 mumol/L antisense ODNs in vitro. 1.5 x 10(6) LCI-D20 cells with or without pretreatment were inoculated into each elevated subcutaneous (s.c) flap in fourteen nude mice, 6 animals for antisense H-ras ODN treated cells, 4 for H-ras non-specific antisense ODN treated cells, the rest 4 for cells without pretreatment.


RESULTS
In in vitro cell culture study, 5-day continuous suppression of H-ras expression by antisense H-ras ODNs resulted in significant inhibition of the proliferation of LCI-D20 cells (t = 31.529, P < 0.01). Cell cycle analysis showed a significant decrease in S phase (36.0 +/- 1.4) and a remarkable increase in G1/G0 fraction (56.7 +/- 1.1) after exposure to antisense H-ras ODNs in comparison with the cells without any treatment (58.5 +/- 0.9, t = 13.519, P < 0.01, 37.4 +/- 0.7, t = 14.802, P < 0.01). In situ end-labeling (ISEL) detection showed that apoptotic cell death was significantly increased in cells with 5-day treatment of antisense H-ras ODNs (34.0% +/- 4.5%) in comparing with cells without treatment (2.5% +/- 1.2%, t = 13. 434, P < 0.01) or treated with non-specific antisense ODNs (4.8% +/- 1.4%, t = 12.453, P < 0.01) at the corresponding time. In in vivo experiment, after six-week observation, tumor growth in antisense H-ras treated animals was significantly retarded in comparison with that of the untreated (t = 3.509, P < 0.01) or nonspecific antisense ODN treated animals (t = 3.452, P < 0.01).


CONCLUSIONS
Specific inhibition of H-ras expression by antisense H-ras ODNs could not only induce apoptotic cell death, inhibit the growth rate of LCI-D20 cells in vitro and in vivo, but also alter in vivo tumorigenesity and metastatic potential of LCI-D20 cells.
Liao
Y
Y
Liver Cancer Institute, Shanghai Medical University.
Tang
Z
Z
Sun
F
F
D000818:Animals; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic; D011905:Genes, ras / Q000276:immunology*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D008175:Lung Neoplasms / Q000556:secondary / Q000628:therapy; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D016376:Oligonucleotides, Antisense; D014407:Tumor Cells, Cultured
D016376:Oligonucleotides, Antisense