Moringa oleifera leaves protein suppresses T-lymphoblastic leukemogenesis via MAPK/AKT signaling modulation of apoptotic activation and autophagic flux regulation

Abstract

Acute lymphoblastic leukemia (T-ALL) is one of the most common hematologic malignancies in children worldwide. Despite advances in chemotherapy in recent years, more than 50% of adult T-ALL patients still experience treatment resistance and relapse/refractory disease. Moringa oleifera, a new food resource in China, has high nutritional value and various pharmacological activities. Moringa oleifera protein is one of the main active ingredients in Moringa oleifera leaves, and some studies have shown that Moringa oleifera protein has antioxidant, anti-inflammatory, antibacterial, anticancer, and other bioactivities; however, the anti-acute lymphoblastic leukemia (T-ALL) activity of the Moringa oleifera protein and its mechanism of action are still unclear. In this study, we used Moringa oleifera leaves protein as the experimental material and used the MTT assay, flow cytometry, and Western blot techniques to study the effects of Moringa oleifera leaves protein on Jurkat cell growth, apoptosis, and the cell cycle in vitro and the underlying molecular mechanism. Moringa oleifera leaves protein inhibited Jurkat cell proliferation; induced apoptosis and cycle arrest; increased autophagy; and inhibited Jurkat cell proliferation by regulating the MAPK/AKT pathway. The results of this study provide new ideas and a theoretical basis for the development of Moringa oleifera leaves protein for the prevention and treatment of leukemia.

Keywords: Jurkat cells; MAPK/AKT signaling pathway; Moringa oleifera leaves protein; autophagy; cell proliferation; immunosuppression.

Figure 1

M. oleifera leaves protein inhibited the proliferation of Jurkat cells. (a) Effect of M. oleifera leaves protein on five cancer cell lines; (b) effect of M. oleifera leaves protein on the human T-lymphoblastic leukemia cell line Jurkat; (c) effect of denatured M. oleifera leaves protein on the viability of Jurkat cells; (d) M. oleifera leaves protein on the viability of GES-1 cells. Effect of M. oleifera leaves protein on Jurkat and GES-1 cells were treated with 30, 60, 90, 120, or 150 μg/mL M. oleifera leaves protein for 24, 48 or 72 h. Values represent the mean ± SEM of the mean; ****P < 0.0001; ***P < 0.001; ns indicates no significant difference. For each group, the data were from 3 separate experiments and 6 parallel samples (n =6).

Figure 2

Effects of M. oleifera leaves protein on the apoptosis of Jurkat cells. (a) Effect of M. oleifera leaves protein on Jurkat cell apoptosis. (b) Early apoptosis. (c) Late apoptosis. (d) Total number of apoptotic cells. Compared with the blank control group (0 μg/mL), the difference between groups is expressed as *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Figure 3

Effects of M. oleifera leaves protein on the Jurkat cell cycle. (a) Effects of M. oleifera leaves protein on the Jurkat cell cycle. (b) Effects of M. oleifera leaves protein on the G1 phase of Jurkat cells. (c) Effects of M. oleifera leaves protein on the S phase of Jurkat cells. (d) Effects of M. oleifera leaves protein on the G2 phase of Jurkat cells. Compared with the blank control group (0 μg/mL), the differences between the groups are expressed as **P < 0.01 and ****P < 0.0001. ns indicates no significant difference.

Figure 4

Effects of M. oleifera leaves protein on Jurkat cell apoptosis and cyclin and autophagy protein expression. (a) Effects of M. oleifera leaves proteins on the expression of apoptosis, cyclin and autophagy proteins in Jurkat cells. (b) Bax/Bcl-2, (c) cleaved CasPase-3, (d) CyclinE1, (e) CyclinD1 and (f) ATG5 statistical analysis of protein expression. Compared with the blank control group (0 μg/mL), the differences between the groups are represented by *P<0.05, **P<0.01, and ***P<0.001. ns indicates no significant difference.

Figure 5

M. oleifera leaves protein regulates the growth mechanism of Jurkat cells. (a) Effects of M. oleifera leaves protein on the expression of pathway-related proteins. (b) -AKT/AKT, (c) p-p38/p38, and (d) p-ERK/ERK ratios were statistically analyzed for protein expression. Compared with the blank control group (0 μg/mL), the differences between the groups are represented by *P<0.05 and **P<0.01. ns indicates no significant difference.