In vitro effects of papaverine on cell proliferation, reactive oxygen species, and cell cycle progression in cancer cells

Gomes, Daniella Anthea de Agrela; Joubert, Anna Margaretha; Visagie, M.H. (Michelle Helen)

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In vitro effects of papaverine on cell proliferation, reactive oxygen species, and cell cycle progression in cancer cells

Gomes, Daniella Anthea de Agrela; Joubert, Anna Margaretha; Visagie, M.H. (Michelle Helen)

URI: https://repository.up.ac.za/handle/2263/85705

Date: 2021-10-22

Abstract:

Papaverine (PPV) is an alkaloid isolated from the Papaver somniferum. Research has shown that PPV inhibits proliferation. However, several questions remain regarding the effects of PPV in tumorigenic cells. In this study, the influence of PPV was investigated on the proliferation (spectrophotometry), morphology (light microscopy), oxidative stress (fluorescent microscopy), and cell cycle progression (flow cytometry) in MDA-MB-231, A549, and DU145 cell lines. Exposure to 150 M PPV resulted in time- and dose-dependent antiproliferative activity with reduced cell growth to 56%, 53%, and 64% in the MDA-MB-231, A549, and DU145 cell lines, respectively. Light microscopy revealed that PPV exposure increased cellular protrusions in MDA-MB-231 and A549 cells to 34% and 23%. Hydrogen peroxide production increased to 1.04-, 1.02-, and 1.44-fold in PPV-treated MDA-MB-231, A549, and DU145 cells, respectively, compared to cells propagated in growth medium. Furthermore, exposure to PPV resulted in an increase of cells in the sub-G1 phase by 46% and endoreduplication by 10% compared to cells propagated in growth medium that presented with 2.8% cells in the sub-G1 phase and less than 1% in endoreduplication. The results of this study contribute to understanding of effects of PPV on cancer cell lines.

Description:

SUPPLEMENTARY MATERIAL 1: FIGURE S1: Spectrophotometry results of crystal violet staining demonstrating the effects of PPV (10–300 M) on proliferation on MDA-MB-231 cells compared to A549- and DU145 cell lines at 24 h, FIGURE S2: Spectrophotometry results of crystal violet staining demonstrating the effects of PPV (10–300 M) on proliferation on MDA-MB-231 cells compared to A549- and DU145 cell lines at 96 h.

SUPPLEMENTARY MATERIAL 2: FIGURE S1: Light microscopy images of cell morphology demonstrating the effects of PPV ((10–150 M) on cell morphology on MDA-MB-231 cells at 48 h at a magnification of 10. TABLE S1: table displaying the effects of papaverine on morphology as percentage change when compared to cells propagated in growth medium on MDA-MB-231 at 48 h. FIGURE S2: Light microscopy images of cell morphology demonstrating the effects of PPV ((10–150 M) on cell morphology on A549 cells at 48 h at a magnification of 10. TABLE S2: table displaying the effects of papaverine on morphology as percentage change when compared to cells propagated in growth medium on A549 at 48 h. FIGURE S3: Light microscopy images of cell morphology demonstrating the effects of PPV ((10–150 M) on cell morphology on DU145 cells at 48 h at a magnification of 10 TABLE S3: table displaying the effects of papaverine on morphology as percentage change when compared to cells propagated in growth medium on DU145 at 48 h. FIGURE S4: Light microscopy images of cell morphology demonstrating the effects of PPV ((10–150 M) on cell morphology on MDA-MB-231 cells at 72 h at a magnification of x10. TABLE S4: table displaying the effects of papaverine on morphology as percentage change when compared to cells propagated in growth medium on MDA-MB-231 at 72 h. FIGURE S5: Light microscopy images of cell morphology demonstrating the effects of PPV ((10–150 M) on cell morphology on A549 cells at 72 h at a magnification of 10. TABLE S5: table displaying the effects of papaverine on morphology as percentage change when compared to cells propagated in growth medium on A549 at 72 h. FIGURE S6: Light microscopy images of cell morphology demonstrating the effects of PPV ((10–150 M) on cell morphology on DU145 cells at 72 h at a magnification of 10. TABLE S6: table displaying the effects of papaverine on morphology as percentage change when compared to cells propagated in growth medium on DU145 at 72 h. FIGURE S7. Light microscopy results demonstrating the effects of ESE-ol used as a positive control on cell morphology.

SUPPLEMENTARY MATERIAL 3; TABLE S1. table displaying the effects of papaverine on oxidative stress as a change of fluorescence intensity relative to the fluorescence intensity of cells propagated in growth medium on MDA-MB-231-, A549- and DU145 cell lines at 48 h. FIGURE S1. Fluorescence staining showing H2O2 production in MDA-MB-231 cells after 48 h. FIGURE S2. Fluorescence staining showing H2O2 production in A549 cells after 48 h. FIGURE S3. Fluorescence staining showing H2O2 production in MDA-MB-231 cells after 48 h. TABLE S2. table displaying the effects of papaverine on oxidative stress as a change of fluorescence intensity relative to the fluorescence intensity of cells propagated in growth medium on MDA-MB-231-, A549- and DU145 cell lines at 72 h. FIGURE S4. Fluorescence staining showing H2O2 production in MDA-MB-231 cells after 72 h. FIGURE S5. Fluorescence staining showing H2O2 production in A549 cells after 72 h. FIGURE S6. Fluorescence staining showing H2O2 production in MDA-MB-231 cells after 72 h.

SUPPLEMENTARY MATERIAL 4: FIGURE S1. Flow cytometry results demonstrating the effects of PPV (10–150 M) on the cell cycle on MDA-MB-231-, A549- and DU145 cells at 48 h. FIGURE S2. Cell cycle progression of MDA-MB-231 cells treated with PPV (10–150 M) at 48 h. FIGURE S3. Cell cycle progression of A549 cells treated with PPV (10–150 M) at 48 h. FIGURE S4. Cell cycle progression of DU145 cells treated with PPV (10–150 M) at 48 h. FIGURE S5. Flow cytometry results demonstrating the effects of PPV (10–150 M) on the cell cycle on MDA-MB-231-, A549- and DU145 cells at 72 h. FIGURE S6. Cell cycle progression of MDA-MB-231 cells treated with PPV (10–150 M) at 72 h. FIGURE S7. Cell cycle progression of A549 cells treated with PPV (10–150 M) at 72 h. FIGURE S8. Cell cycle progression of DU145 cells treated with PPV (10–150 M) at 72 h.

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