Heat stress induced apoptosis in BC-8 cells derived from AK-5 tumor involves downregulation of Bcl-2 and generation of reactive oxygen species.

BC-8, a rat histiocytoma undergoes apoptosis after heat shock, which is due to lack of an effective heat shock response. Heat shock induced generation of free radicals, which in turn are involved in the induction of apoptotic death in BC-8 cells. Treatment of BC-8 cells with N-acetylcysteine partially inhibited the heat induced apoptosis. Introduction of Bcl-2 gene in these cells did not protect them from apoptotic death, whereas transfection with hsp-70 gene did render these cells resistant to heat induced apoptosis transiently. Heat shock also downregulated the expression of Bcl-2 and p53 in these cells. These observations suggested that the heat shock induced apoptosis was mediated through reactive oxygen species and controlled upstream of Bcl-2 check point.

Differential cytotoxic effects of Annona squamosa seed extracts on human tumour cell lines: role of reactive oxygen species and glutathione.

Annonaceous acetogenins are a new class of compounds that have been reported to have potent pesticidal, parasiticidal, anti-microbial, cell growth inhibitory activities. In this study, organic and aqueous extracts from the defatted seeds of Annona squamosa (custard apple) were tested on different human tumour cell lines for antitumoural activity. While organic and aqueous extracts induced apoptosis in MCF-7 and K-562 cells, they failed to do so in COLO-205 cells. Treatment of MCF-7 and K-562 cells with organic and aqueous extracts resulted in nuclear condensation, DNA fragmentation, induction of reactive oxygen species (ROS) generation and reduced intracellular glutathione levels. In addition downregulation of Bcl-2 and PS externalization by Annexin-V staining suggested induction of apoptosis in MCF-7 and K-562 cells by both the extracts through oxidative stress. On the contrary, COLO-205 cells showed only PS externalization but no change in ROS and glutathione levels. These observations suggest that the induction of apoptosis by A. squamosa extracts can be selective for certain types of cancerous cells.

Induction of apoptosis in AK-5 cells by rotenone involves participation of caspases.

AK-5 tumour cells undergo apoptosis after treatment with rotenone an electron transport inhibitor and oligomycin which inhibits mitochondrial ATPases. Apoptotic process involves the induction of caspases 2 and 3, whereas caspase 1 does not seem to be participating in rotenone/oligomycin induced apoptosis. DEVD which is a specific inhibitor of caspase 3, inhibited apoptosis in AK-5 cells. We have also observed a significant lowering of intracellular pH in AK-5 cells which are induced into the apoptotic process by rotenone. These results suggest an important role for mitochondrial electron transport in the induction of apoptosis in AK-5 tumour cells.

Increased apoptosis in AK-5 tumor cells after transfection with wild type p53 gene.

AK-5 tumor cell death is mediated through necrosis and apoptosis. The apoptotic activity in AK-5 cells has been studied after introduction of wild-type p53 gene. Apoptotic activity both in vitro and in vivo is significantly increased in p53 transfected cells. Similarly there is fragmentation of tumor cell DNA after undergoing apoptosis. These observations suggest an active participation of p53 in inducing apoptosis in AK-5 tumor cells.

Conjugate formation between effector and target as a function of cytotoxicity in antibody-dependent killing of AK-5 tumor.

The present study was carried out to mechanistically view a possible correlation between the process of, conjugate formation and its relation to target cell death. AK-5 killing is mediated by CD8+ natural killer cells through ADCC. Immune effectors on exposure to antibody primed AK-5 tumor cells formed tight conjugates. Ability of various cell types (NK, T, monocytes and macrophages) to form conjugates was evaluated. Marked increase in the number of NK cells binding to the target as compared to the other cell types was observed. Cytotoxicity of free and bound effectors against antibody tagged AK-5 cells demonstrated a reduced cytotoxic ability of the former in contrast to a significantly high lytic potential of the bound effectors. The results highlight the requirement for priming of NK cells which mediate killing of AK-5 tumor and provide additional evidence that formation of stable conjugates acts as the first signal for triggering lymphocyte activation and effective target cell lysis.