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

Abstract

Introduction Nutlin‐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 Methods Rat‐derived bone marrow‐derived MSCs (BMSCs) were cultured and treated with different concentrations (5, 10, 25, 50, and 100 μM) and times (24, 48, and 72 hr) 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. Results The viability of BMSCs in the treated cells with concentrations of 100 μM at 24 hr, 50 and 100 μM at 48 hr, and in all concentrations at 72 hr was significantly (p < 0.05) low. The apoptotic index showed that the TUNEL‐positive BMSCs were significantly higher in concentrations of 25 and 50 μM in comparison to control group ( p < 0.05) and augmented in a dose‐dependent manner. Annexin‐V‐PI staining showed after 72 hr of incubation, there was a slight dose‐dependent increase in total apoptotic cells at 10 and 25 μM of Nutlin‐3, but a massive significant increase at 50 μM. Conclusion Here, 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.

Resistance of human primary mesenchymal stem cells to cytotoxic effects of nutlin‐3 in vitro

“…These cells were found to be relatively resistant to nutlin-3 in rats 33 and human BMSCs. These cells were found to be relatively resistant to nutlin-3 in rats 33 and human BMSCs.…”

“…[9][10][11][12][13][14] Moreover, research suggests that nutlin-3 induces apoptosis and suppresses cell growth in the absence of wild-type TP53 through the TP53 homolog of p73, 15,16 and sensitizes TP53-defective cancer cells to different anticancer agents, such as arsenic trioxide, 17 radiation, 18 and doxorubicin. 33 The present research seeks to investigate the effectiveness of nutlin-3 in the proliferation and survival of MSCs obtained from human bone marrow (BMSCs). 20 Preclinical studies found nutlin-3 to present treatment potential for human cancer cells with wild-type TP53.…”

“…The effect of nutlin‐3 on human's MSCs, however, should be identified. Rat‐derived BMSCs were found to be relatively resistant to nutlin‐3 . The present research seeks to investigate the effectiveness of nutlin‐3 in the proliferation and survival of MSCs obtained from human bone marrow (BMSCs).…”

Palmitic Acid Methyl Ester Induces G2/M Arrest in Human Bone Marrow-Derived Mesenchymal Stem Cells via the p53/p21 Pathway

“…Mdm2 has been demonstrated to play a major role in regulating hematopoietic stem cell survival [57]. Moreover, previous studies have shown that Mdm2 inhibitors can inhibit cell proliferation in rBM-MSCs and hBM-MSCs [20, 21]. Dysfunction of the Mdm2/p53 axis has been linked to cell proliferation and apoptosis.…”

“…In normal cells, p53 is kept at low levels by murine double minute 2 (Mdm2), which can bind to p53 and act as p53 ubiquitin ligase that negatively regulates p53 function in several cellular pathways, such as cell cycle and apoptosis. Previous studies have shown that the Mdm2 inhibitor can decrease cell proliferation in rat BM-MSCs (rBM-MSCs) and hBM-MSCs [20, 21]. However, the mechanism underlying PAME-induced p53 stabilization and consequent cell cycle arrest in hBM-MSCs has not been elucidated.…”