Establishment and characterization of a human hepatocellular carcinoma cell line EHBC-512 / 第二军医大学学报

ABSTRACT

Objective: To establish a new hepatocellular carcinoma cell line EHBC-512 and to analyze its biological property, so as to lay a foundation for future study. Methods: The specimen of liver cancer was resected during operation and was primary cultured. The morphology of cells was observed under optical and electron microscope. The cell cycle was examined with flow cytometry; the chromosome karyotype was also analyzed. The expression of AFP in the culture supernatant was detected by immunofluorescence and chemoluminescence. The cultured cells were implanted into nude mice for observation of tumor formation. Results: The cultured cell line had the character of liver cancer under light microscope, and had rich mitochondria, with obvious nuclear heteromorphism and a chromosome number of 110-120. The doubling time of the cells was 48 h, and the adherence rate was 90%. Immunofluorescence showed that the cells were positive of AFP and CK18 and negative of CEA and CK19. The production of AFP was higher and the content of AFP was higher than 1210 μg/L. The histology of the exografts was similar to that of the primary tumor. Conclusion: We have successfully established a new hepatocellular cell line EHBC-512, which can stably secrete AFP in vitro, providing a novel tool for liver cancer research.