Protein degradation in lysosomes from chemically induced malignant rat hepatoma

Hepatocellular carcinomas were induced in rat liver by exposing the animals to diethylnitrosamine and 2-acetylaminofluorene in combination with partial hepatectomy. Light and electron microscopy demonstrated that the general appearance of the tumour tissue was that of highly differentiated malignant hepatocytic cells. Morphometrically there was a difference between normal and malignant cells in that the entire lysosomal apparatus was twice as large in malignant cells as in normal cells. This was mainly due to an increase in the fractional volume of autophagic vacuoles. A total lysosomal fraction (dense bodies and autophagic vacuoles) was isolated and characterized from both control and tumour livers. Marker enzyme analysis showed that the lysosomal enzyme activities were significantly lower in malignant liver tissue. Injection of leupeptin, an inhibitor of cathepsins B, H, and L, into rats did not increase the fractional volume of autophagic vacuoles in tumour tissue as much as in normal liver tissue. The proteolytic rate was lower in the lysosomal fraction from hepatoma cell tissue compared with the lysosomal fraction from normal cell tissue. This could conceivably be due to the lower activities of lysosomal enzymes. However, if the recovery of lysosomes is taken into account no clear-cut difference in lysosomal proteolysis between control and malignant liver tissue was noted. Accordingly, in malignant liver tissue a proteolytic balance is obtained characterized by an increased fractional volume of AVs and lower rate of protein degradation in individual lysosomes.