A continuous adherent cell line was established from a hepatocellular carcinoma of an HBsAg-positive Italian male. This cell line, designated Tong/HCC, has been grown in a hormone-supplemented medium for more than 18 months. The cell line secretes HBsAg, a-fetoprotein, albumin and al-antitrypsin. a-Fetoprotein production is enhanced by the addition of hydrocortisone and appears to be glumorticoid concentration-dependent. The concentrates of the supernatant from the cell cultures and cell lysates were negative when tested for HBeAg. The cell culture medium was negative for hepatitis B virus DNA when tested by dot-blot hybridization. However, hepatitis B virus DNA was found to be integrated in the chromosomal DNA by Southern blot analysis. At least five different integration sites were identified, and no free hepatitis B virus DNA was observed.
The modal chromosome number was 64, and a translocationon Chromosome 16 was consistently noted. H L A typing revealed sites for A3, Aw24, Bw34 and Cwl.
Recent studies (1-4) have suggested a close association between hepatitis B virus (HBV) infection and the development of hepatocellular carcinoma (HCC). Several HCC cell lines have been established, some of which produce HBsAg (5-8) while others do not (7, 9, 10). These cell lines have potential importance in studying the postulated oncogenicity of HEW. This paper reports the characteristics of a new HCC cell line designated Tong/HCC. The cell line was derived from a 59-year-old male Italian whose serum was positive for HBsAg, HBeAg, HBV DNA, anti-HBc, and had high a-fetoprotein ( A m ) levels. Both HBsAg and AFP are produced by the cell line.
Methods
Source of firnor. The tumor was obtained from a fresh autopsy sample of HCC from a 59-year-old Italian male.
n m u e Culture. Using sterile technique, the tumor tissue was cut into pieces of less than 1 mm' in size. The tissue pieces were washed with Hanks' balanced salt solution, plated in 60 mm Coming tissue culture dishes, and covered with 4 ml of