1. ¹Department of Medicine I, Division: Institute of Cancer Research, Medical University of Vienna, Borschkegasse 8a, Vienna A-1090, Austria
2. ²Department for Surgery, Medical University of Vienna, Währinger Gürtel 18-20, Vienna A-1090, Austria
3. ³Institute for Clinical Pathology, Medical University of Vienna, Währinger Gürtel 18-20, Vienna A-1090, Austria
4. ⁴Department of Medicine I, Division for Hematology, Medical University of Vienna, Währinger Gürtel 18-20, Vienna A-1090, Austria
5. ⁵Institute of Immunology, Medical University of Vienna, Borschkegasse 8a, Vienna A-1090, Austria
6. ⁶Department of Pathology, Medical University of Graz, Auenbruggerplatz 25, Graz A-8036, Austria

Correspondence: Dr B Grasl-Kraupp, E-mail: bettina.grasl-kraupp@meduniwien.ac.at

⁷These authors contributed equally to this work.

Revised 17 April 2008; Accepted 17 April 2008.

Abstract

To enable detailed analyses of cell interactions in tumour development, new epithelial and mesenchymal cell lines were established from human hepatocellular carcinoma by spontaneous outgrowth in culture. We obtained several hepatocarcinoma (HCC)-, B-lymphoblastoid (BLC)-, and myofibroblastoid (MF)-lines from seven cases. In-depth characterisation included cell kinetics, genotype, tumourigenicity, expression of cell-type specific markers, and proteome patterns. Many functions of the cells of origin were found to be preserved. We studied the impact of the mesenchymal lines on hepatocarcinogenesis by in vitro assays. BLC- and MF-supernatants strongly increased the DNA replication of premalignant hepatocytes. The stimulation by MF-lines was mainly attributed to HGF secretion. In HCC-cells, MF-supernatant had only minor effects on cell growth but enhanced migration. MF-lines also stimulated neoangiogenesis through vEGF release. BLC-supernatant dramatically induced death of HCC-cells, which could be largely abrogated by preincubating the supernatant with TNF-antiserum. Thus, the new cell lines reveal stage-specific stimulatory and inhibitory interactions between mesenchymal and epithelial tumour cells. In conclusion, the new cell lines provide unique tools to analyse essential components of the complex interplay between the microenvironment and the developing liver cancer, and to identify factors affecting proliferation, migration and death of tumour cells, neoangiogenesis, and outgrowth of additional malignancy.