Original ContributionInduction of caspase-independent apoptotic-like cell death of mouse mammary tumor TA3Ha cells in vitro and reduction of their lethality in vivo by the novel chemotherapeutic agent GIT-27NO
Section snippets
Animals
Female strain A inbred mice, 5–6 weeks of age, were obtained from Charles River (Lecco, Italy) and kept under standard (non-SPF) laboratory conditions with free access to food and water. The handling of animals and the study protocol were in accordance with international guidelines and approved by the local Institutional Animal Care and Use Committee.
Reagents and cells
Fetal calf serum (FCS), RPMI 1640, phosphate-buffered saline (PBS), dimethyl sulfoxide (DMSO),
GIT-27NO suppresses TA3Ha cell growth in vitro; participation of the NO liberated from the drug
TA3Ha cells were treated with various concentrations of either GIT-27NO or its parental compound VGX-1027 for 24 h. The results of the MTT assay, presented in Fig. 1A, clearly indicate that GIT-27NO, in contrast to VGX-1027, dramatically decreased the number of viable cells in the cultures. The observed effects coincided with significant cellular uptake of drug-generated NO as early as 2 h after the start of incubation (Fig. 1B). A dose-dependent increase in nitrite accumulation in the cell
Discussion
We have demonstrated that the ascitic tumor induced by the TA3Ha cell line is sensitive to GIT-27NO treatment and we provide novel information regarding the mode of action of GIT-27NO in this setting. High expression of hyaluron receptors and mucins such as carbohydrate glycoprotein–epiglycanan on the membranes of TA3Ha cells confers on them an extremely invasive phenotype capable of escaping specific immune responses [18]. Despite these complex biochemical characteristics of TA3Ha cells,
Acknowledgments
This work was supported by the Serbian Ministry of Science (Grant 143029). We thank Dr. Gianni Garotta (Ganial Immunotherapeutics, Inc., Wilmington, DE, USA) for providing GIT-27NO and Igor Golic (Faculty of Biology, University of Belgrade, Belgrade, Serbia) for performing electron microscopy.
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