The antiproliferative effect of Moringa oleifera crude aqueous leaf extract on cancerous human alveolar epithelial cells - PubMed (original) (raw)

The antiproliferative effect of Moringa oleifera crude aqueous leaf extract on cancerous human alveolar epithelial cells

Charlette Tiloke et al. BMC Complement Altern Med. 2013.

Abstract

Background: The incidence of lung cancer is expected to increase due to increases in exposure to airborne pollutants and cigarette smoke. Moringa oleifera (MO), a medicinal plant found mainly in Asia and South Africa is used in the traditional treatment of various ailments including cancer. This study investigated the antiproliferative effect of MO leaf extract (MOE) in cancerous A549 lung cells.

Methods: A crude aqueous leaf extract was prepared and the cells were treated with 166.7 μg/ml MOE (IC50) for 24 h and assayed for oxidative stress (TBARS and Glutathione assays), DNA fragmentation (comet assay) and caspase (3/7 and 9) activity. In addition, the expression of Nrf2, p53, Smac/DIABLO and PARP-1 was determined by Western blotting. The mRNA expression of Nrf2 and p53 was assessed using qPCR.

Results: A significant increase in reactive oxygen species with a concomitant decrease in intracellular glutathione levels (p < 0.001) in MOE treated A549 cells was observed. MOE showed a significant reduction in Nrf2 protein expression (1.89-fold, p < 0.05) and mRNA expression (1.44-fold). A higher level of DNA fragmentation (p < 0.0001) was seen in the MOE treated cells. MOE's pro-apoptotic action was confirmed by the significant increase in p53 protein expression (1.02-fold, p < 0.05), p53 mRNA expression (1.59-fold), caspase-9 (1.28-fold, p < 0.05), caspase-3/7 (1.52-fold) activities and an enhanced expression of Smac/DIABLO. MOE also caused the cleavage and activation of PARP-1 into 89 KDa and 24 KDa fragments (p < 0.0001).

Conclusion: MOE exerts antiproliferative effects in A549 lung cells by increasing oxidative stress, DNA fragmentation and inducing apoptosis.

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Figures

Figure 1

Figure 1

Oxidative stress induced by MOE on A549 cells. An increase in MDA levels (lipid peroxidation) (A) and decreased intracellular GSH levels (B) in MOE treated cells (**p < 0.001).

Figure 2

Figure 2

Comet assay images of control and MOE treatments for 24 h. DNA damage was higher in cells exposed to MOE (B) then control cells (A) (100×, ***p < 0.0001).

Figure 3

Figure 3

MOE regulating protein expression in A549 cells. Differential expression of Nrf2, p53, Smac/DIABLO, PARP-89 KDa and 24 KDa fragment in A549 cells after treatment with MOE for 24 h.

Figure 4

Figure 4

The effect of MOE on mRNA expression. MOE regulated the Nrf2 and p53 mRNA expression in A549 cells after treatment for 24 h (**p < 0.001).

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