VERNONIA AMYGDALINA DELILE EXHIBITS A POTENTIAL FOR THE TREATMENT OF ACUTE PROMYELOCYTIC LEUKEMIA - PubMed (original) (raw)

VERNONIA AMYGDALINA DELILE EXHIBITS A POTENTIAL FOR THE TREATMENT OF ACUTE PROMYELOCYTIC LEUKEMIA

Clement G Yedjou et al. Glob J Adv Eng Technol Sci. 2018 Aug.

Abstract

The World Health Organization (WHO) has been on front line to encourage developing countries to identify medicinal plants that are safe and easily available to patients. Traditional medicine represents the first-treatment choice for the healthcare of approximately 80% of people living in developing countries. Also, its use in the United States has increased by 38% during within the last decade of the 20th century alone. Therefore, the aim of the present study was to explore the efficacy of a medicinal plant, Vernonia amygdalina Delile (VAD), as a new targeted therapy for the management of acute promyelocytic leukemia (APL), using HL-60 cells as a test model. To address our specific aim, HL-60 promyelocytic leukemia cells were treated with VAD. Live and dead cells were determined by acridine orange and propidium iodide (AO/PI) dye using the Cellometer Vision. The extent of DNA damage was evaluated by the comet assay. Cell apoptosis was evaluated by flow cytometry assessment. Data obtained from the AO/PI assay indicated that VAD significantly reduced the number of live cells in a dose-dependent manner, showing a gradual increase in the loss of viability in VAD-treated cells. We observed a significant increase in DNA damage in VAD-treated cells compared to the control group. Flow cytometry data demonstrated that VAD induced apoptosis in treated cells compared to the control cells. These results suggest that induction of cell death, DNA damage, and cell apoptosis are involved in the therapeutic efficacy of VAD. Because VAD exerts anticancer activity in vitro, it would be interesting to perform clinical trials to confirm its effectiveness as an anticancer agent towards the treatment of APL patients.

Keywords: DNA damage; Vernonia amygdalina Delile; apoptosis; promyelocytic leukemia (HL-60) cells.

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Conflict of interest statement

Conflicts of Interest The authors declare no conflict of interest

Figures

Figure 1:

Figure 1:

Photo of VAD leaves taken in Bangou, West Cameroon on May 10, 2012 by Clement G. Yedjou. This photo shows how VAD leaves are processed for food: (1) Whole leaves; (2) Removal of stem and leaf veins; (3) Leaves cut in small pieces, ready to be washed several times to remove the bitter taste. After washing, it is cooked using different recipes. The production of bitter-leaves from VAD contributes to the food security and economic development in Cameroon, and helps to sustain the environment.

Figure 2:

Figure 2:

Bright field images (left) and fluorescent images (right) of HL-60 cells exposed to VAD for 24 h. Figure 2 shows HL-60 cells untreated (A-control) and HL-60 cells treated with VAD at 125 µg/mL (B), 250 µg/mL (C), and 500 µg/mL (D). Live cells (green fluorescent) and dead cells (red fluorescent) were determined based on the acridine orange and propidium iodide assay using the Cellometer Vision. The cell diameters vary from 8 to 12 µm with the average of 10 µm.

Figure 3:

Figure 3:

Antiproliferative effect of VAD to HL-60 promyelocytic leukemia cells. HL-60 cells were cultured with increasing doses of VAD (0, 125, 250, and 500 µg/mL) for 24 has indicated in the Materials and Methods. Cell viability was determined based on the acridine orange and propidium iodide assay. Each point represents a mean ± SD of 3 experiments with 6 replicates per dose. *Significantly different (p<0.05) from the control, according to the Dunnett’s test.

Figure 4:

Figure 4:

Representative SYBR Green Comet assay images of untreated (A-control) and VAD. HL-60 promyelocytic leukemia cells treated with VAD at 125 µg/mL (B), 250 µg/mL (C), and 500 µg/mL (D). A total volume of 50 µL from 1 × 105 cells/mL was used for each treatment as indicated in the Materials and Methods. Untreated cells (A) showed absence of DNA migration in cultured HL-60 cells while cells treated with VAD (B, C, and D) showed clear migration of DNA from the head to tail regions.

Figure 5:

Figure 5:

Bar graph showing the percentage of DNA cleavage (A) and tail length (B) in untreated and VAD-treated HL-60 cells. A total volume of 50 µL from 1 × 105 cells/mL was used for each treatment as indicated in the Materials and Methods. Each point represents mean ± SD of 3 independent experiments. *Significantly different (p < 0.05) from the control, according to the Dunnett’s test.

Figure 6:

Figure 6:

Representative flow cytometry analysis data from Annexin V/PI staining. The histograms show a comparison of the distribution of annexin V/PI negative cells (M1) and annexin V/PI positive cells (M2) of VAD-treated cells for 24 h. A-control; B-125μg/mL VAD; C-250μg/mL VAD; and D-500μg/mL VAD.

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