The effects of Nutlin-3 on morphology, cellular proliferation, and apoptosis in rat primary mesenchymal stem cells

IntroductionNutlin-3 is a powerful antagonist of murine double minute 2/p53 interaction demonstrating promising therapeutic anticancer activity, which has not been clinically approved yet. Mesenchymal stem cells (MSCs) are an important part of the bone marrow niche and support regeneration and proliferation of hematopoietic stem cells after exposure to myelotoxic anticancer agents; however, the effect of Nutlin-3 compounds on MSCs themselves remains to be elucidated. Materials and MethodsRat-derived bone marrow-derived MSCs (BMSCs) were cultured and treated with different concentrations (5, 10, 25, 50, and 100M) and times (24, 48, and 72hr) of Nutlin-3. The microculture tetrazolium test, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and propidium iodide and annexin-V assays, and quantitative real-time reverse-transcription polymerase chain reaction were performed to assess the effects of Nutlin-3 on the cell viability, proliferation, and apoptosis in BMSCs. ResultsThe viability of BMSCs in the treated cells with concentrations of 100M at 24hr, 50 and 100M at 48hr, and in all concentrations at 72hr was significantly (p<0.05) low. The apoptotic index showed that the TUNEL-positive BMSCs were significantly higher in concentrations of 25 and 50M in comparison to control group (p<0.05) and augmented in a dose-dependent manner. Annexin-V-PI staining showed after 72hr of incubation, there was a slight dose-dependent increase in total apoptotic cells at 10 and 25M of Nutlin-3, but a massive significant increase at 50M. ConclusionHere, we show that rat BMSCs are relatively resistant to Nutlin-3; however, further in vivo data with long-term exposure may help to corroborate our findings.