دراسة الخصائص النسجية و الخلوية لخط خلايا سرطان الكبد

Study of histological and cellular characteristics For Hepatocellular Carcinoma cell line

A new liver cancer cell line Hepatic Cancer Ahmed Majeed (HCAM) has been characterized; it is considered as a useful tool in liver cancer research. The HCAM cell line was established from the primary tumor of a white swiss albino male mouse with spontaneous diffuse hepatocellular carcinoma. The current study in different experiments aimed to characterize our cancer cell line, which are:

A morphological study was carried out by staining the cells with hematoxylin and eosin dyes. They were polygonal epithelial like cells with central nucleus and multi-nuclei, further to the clear vacuoles in cytoplasm. In addition, Studying two growth curves for different cells numbers of HCAM cell line (10,000 cells/1ml and 100,000 cells/1ml), The lag phase was identical for the curves (24 hour), while log phase ranging from 24 hour to 144 hour and the decline phase was 240 hour. We also calculated the population doubling time that has been 54 hour for 10,000 cells/1ml and 47 hour for 100,000 cells/1ml.

The physiological study aimed to characterize HCAM tumor cells in eighth passage by studying the high activity of liver enzymes (Alanine transamminase, Aspartate aminotransferase, Gamma glutamyl transferase, Lactic dehydrogenase and C-reactive protein) U/L (P ≤ 0.05) in tumor cell line comparing to normal liver cells from embryo and adult mice. High values of tumor marker proteins (Alpha-fetoprotein, Carcinoembryonic antigen and Carbohydrate antigen) U/L (P ≤ 0.05) were also found in HCAM cell line compared with normal hepatocytes.

Immunohistochemistry examination was done for the tumor cells grown in tissue culture slides to detect the presence of some immune enzymes: Glutathione S-transferase Mu 1 (GSTM1) gives negative result, Glutathione S-transferase theta 1 (GSTT1) gives week positive result and to test P53 protein that gives clear positive result.

The Cytogenetic study showed chromosomal aberrations with many numerical changes among the tumor cells and abnormal structures (Metacentric chromosomes, Breakpoints in the arms of chromosomes and abnormal chromosomal length) gives chromosomes with unknown origin called marker chromosome.

The Genetic study of HCAM cell line by using traditional polymerase chain reaction showed positive result for Mouse Glyceraldehyde 3-phosphate dehydrogenase gene and negative result for Human Glyceraldehyde 3-phosphate dehydrogenase gene. Conclusion: the hepatic cancer cell line showing high morphological, physiological, Immunohistochemical and genetically activities and this make it useful as liver cancer model.