Published in Cancer Detection and Prevention 1998; 22(Supplement 1).

Approach to inhibit the growth of human hepatocellular carcinoma derived cell lines by transforming growth factor alpha anti-sense DNA oligonucleotide

S. Kira MD¹,ÊT. Nakanishi MD ², M. Kitamoto MD ², G.Kajiyama MD ²

AIM: Previous studies have indicated that transforming growth factor alpha (TGF-α) play an important role in liver regeneration and hepatocarcinogenesis. We also have reported that TGF-α and EGFR was overexpressed in hepatocellular carcinoma(HCC)( Liver 1997; 17:177-182). To better understand the role of TGF-α in development of HCC, we investigated the specific inhibition of TGF-α gene by TGF-α DNA anti-sense phosphorothioate oligonucleotides (S-ODN, Oncogene Science Co.) in HCC derived cell lines. METHODS: 1) Human HCC derived cells (Hep G2,HLF)were plated at 1.0x104cells/well in 96 well plates and grown in 100ul of WEM supplemented with 10% FCS. Human TGF-α S-ODN and control oligonucleotides were added directly into the medium 48hrs after plating cells.3H thymidine incorporation was measured 24hrs after addition.2)TGF-α in culture medium was measured by using enzyme-linked immunosorbent assay kit (Oncogene Science Co.). RESULTS: 1) 3H thymidine incorporation ratio to non addition controls was lower in TGF-α S-ODN than that in control oligonucleotides. 2) TGF-α in culture medium was 6.71 +/-1.57pg/24hr/g protein (HepG2), 4.99 +/-1.58pg/24hr/g protein (HLF). CONCLUSIONS: These results suggest that TGF-α could be an autocrine factor in hepatocellular carcinoma derived cell lines and TGF-α may play a role in development of hepatocellular carcinoma.

KEY WORDS: TGF-alpha, Anti-sense DNA oligonucleotide, human hepatocellular carcinoma, human hepatocellular carcinoma, .

Paper presented at the International Symposium on the Impact of Biotechnology on Prediction, Prevention and Treatment of Cancer; Nice, France; October 24 - 27, 1998; in the section on Angiogenesis & Inhibition.

http://www.cancerprev.org/Journal/Issues/22/101/36/2390