Implication of caspase-3 and granzyme B expression and activity in spleenocytes of ehrlich ascites carcinoma mice subjected to immunotherapy

N Am J Med Sci. 2011 Aug;3(8):371-7. doi: 10.4297/najms.2011.3371.

Abstract

Background: Caspase-3 and granzyme B were claimed as apoptotic manipulative enzymes.

Aims: The present study was to determine the enzymes expression and activity in cancer and cancer immune therapeutic status and the possible association to cancer common pathological signs targeting the improvement of therapeutic conditions.

Material and methods: Mice were immunized with cell lyaste or cell lysate + CKI in the right thigh and challenged with live cells of ehrlich ascites carcinoma (EAC) in the left thigh. The expression and activity of both enzymes in the spleenocytes derived from different subjects (normal, EAC and cell lysate or cell lysate + CKI immunized mice) after cultured with EAC viable cells were determined by colorimetric assay and western blot analysis. In addition, the subjects DNA ladder and serum metalloproteases (MMP 2 and 9) zymography were observed.

Results: The experimental data revealed over expression of caspase3 and granzyme B in the groups of cell lysate or cell lysate + CKI immunized mice compared to control while down expression were recorded in the EAC subject. The over expression of the 2 enzymes were accompanied with increases in the activities of caspase3 and granzyme B, changes in DNA fragmentation and inhibition of metalloproteases.

Conclusion: It could be suggested that, the parameter estimation within the present experimental framework could identify the efficiency of therapeutic vaccine protocols and elucidate the impact of CKI adjuvant with vaccines therapy.

Keywords: CKI; DNA ladder; Enzyme expression; MMPs; apoptosis.