The main treatment of Acute Myeloid Leukemia in children is chemotherapy, which can lead to side effects. Recently, a new isoform of human MnSOD was isolated and obtained in a synthetic recombinant form (rMnSOD). This isoform can kill T-ALL cells, without cytotoxic effects on healthy cells (1). The rMnSOD is characterized by the presence of a leader peptide, which allows the protein to enter cells through oestrogen receptors unlike the wild type form. The selective oncotoxic activity of rMnSOD is due to an increase in the level of H2O2 in cancer cells, which contain lower levels of catalase than healthy cells (2). In this study we analyzed the rMnSOD effects on Kasumi-1 cell line. Methods Detection of oestrogen receptors (ER1 and ER2) was obtained by qPCR assay. Cell viability and apoptosis were analyzed by flow cytometry through Annexin V assay after 1 µM, 1.5µM and 2µM rMnSOD treatments. Apoptotic fragmentations were demonstrated by confocal imaging. Mitochondria were detected by MitoTracker® Red CMXRos. Results A very high amount of both ER1 and ER2 was detected in Kasumi-1 cells. Annexin V analysis showed that 2 µM rMnSOD treatment induced apoptosis in cells (about 50%). Confocal microscopy showed typical alterations of apoptosis, following 24h treatment of 1.5μM and mainly 2μM rMnSOD, such as nuclear fragmentations and apoptotic-bodies. The internalization of rMnSOD in mitochondria was demonstrated by its colocalization with MitoTracker, following 6h incubation. In conclusion, rMnSOD exerts toxic activity against leukemia cell line. Further observations of rMnSOD effects on AML M2 pediatric patient cells are in progress. References 1.Pica A et al. J. Cell Physiol 2015, 230(5):1086-93. 2.Borrelli A et al. Nature Sci Rep 2016, 4;6:18691.
Apoptotic Death of M2 acute myeloid leukemia cells by ROS Production / Manca, Rosa; Pollio, MARIA LAURA; Toriello, M.; Miceli, R.; Camerlingo, R.; Borrelli, A.; Mancini, A.; Pica, Alessandra. - In: EUROPEAN JOURNAL OF HISTOCHEMISTRY. - ISSN 2038-8306. - 61:2(2017), p. 8. [10.4081/ejh.2017.2844]
Apoptotic Death of M2 acute myeloid leukemia cells by ROS Production
MANCA, ROSA;POLLIO, MARIA LAURA;PICA, ALESSANDRA
2017
Abstract
The main treatment of Acute Myeloid Leukemia in children is chemotherapy, which can lead to side effects. Recently, a new isoform of human MnSOD was isolated and obtained in a synthetic recombinant form (rMnSOD). This isoform can kill T-ALL cells, without cytotoxic effects on healthy cells (1). The rMnSOD is characterized by the presence of a leader peptide, which allows the protein to enter cells through oestrogen receptors unlike the wild type form. The selective oncotoxic activity of rMnSOD is due to an increase in the level of H2O2 in cancer cells, which contain lower levels of catalase than healthy cells (2). In this study we analyzed the rMnSOD effects on Kasumi-1 cell line. Methods Detection of oestrogen receptors (ER1 and ER2) was obtained by qPCR assay. Cell viability and apoptosis were analyzed by flow cytometry through Annexin V assay after 1 µM, 1.5µM and 2µM rMnSOD treatments. Apoptotic fragmentations were demonstrated by confocal imaging. Mitochondria were detected by MitoTracker® Red CMXRos. Results A very high amount of both ER1 and ER2 was detected in Kasumi-1 cells. Annexin V analysis showed that 2 µM rMnSOD treatment induced apoptosis in cells (about 50%). Confocal microscopy showed typical alterations of apoptosis, following 24h treatment of 1.5μM and mainly 2μM rMnSOD, such as nuclear fragmentations and apoptotic-bodies. The internalization of rMnSOD in mitochondria was demonstrated by its colocalization with MitoTracker, following 6h incubation. In conclusion, rMnSOD exerts toxic activity against leukemia cell line. Further observations of rMnSOD effects on AML M2 pediatric patient cells are in progress. References 1.Pica A et al. J. Cell Physiol 2015, 230(5):1086-93. 2.Borrelli A et al. Nature Sci Rep 2016, 4;6:18691.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.