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DNA fragmentation and apoptosis induced by safranal in human prostate cancer cell line - PubMed

DNA fragmentation and apoptosis induced by safranal in human prostate cancer cell line

Saeed Samarghandian et al. Indian J Urol. 2013 Jul.

Abstract

Objectives: Apoptosis, an important mechanism that contributes to cell growth reduction, is reported to be induced by Crocus sativus (Saffron) in different cancer types. However, limited effort has been made to correlate these effects to the active ingredients of saffron. The present study was designed to elucidate cytotoxic and apoptosis induction by safranal, the major coloring compound in saffron, in a human prostate cancer cell line (PC-3).

Materials and methods: PC-3 and human fetal lung fibroblast (MRC-5) cells were cultured and exposed to safranal (5, 10, 15, and 20 μg/ml). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to assess cytotoxicity. DNA fragmentation was assessed by gel electrophoresis. Cells were incubated with different concentrations of safranal, and cell morphologic changes and apoptosis were determined by the normal inverted microscope, Annexin V, and propidium iodide, followed by flow cytometric analysis, respectively.

Results: MTT assay revealed a remarkable and concentration-dependent cytotoxic effect of safranal on PC-3 cells in comparison with non-malignant cell line. The morphologic alterations of the cells confirmed the MTT results. The IC50 values against PC-3 cells were found to be 13.0 ΁ 0.07 and 6.4 ΁ 0.09 μg/ml at 48 and 72 h, respectively. Safranal induced an early and late apoptosis in the flow cytometry histogram of treated cells, indicating apoptosis is involved in this toxicity. DNA analysis revealed typical ladders as early as 48 and 72 h after treatment, indicative of apoptosis.

Conclusions: Our preclinical study demonstrated a prostate cancer cell line to be highly sensitive to safranal-mediated growth inhibition and apoptotic cell death. Although the molecular mechanisms of safranal action are not clearly understood, it appears to have potential as a therapeutic agent.

Keywords: Apoptosis; DNA; PC-3; cytotoxicity; safranal.

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Conflict of interest statement

Conflict of Interest: None declared.

Figures

Figure 1
Figure 1

Effect of safranal on cell viability of PC-3 and MRC-5 cells. Cells were treated with different concentrations of safranal for 24, 48, and 72 h. Viability was quantitated by MTT assay. Results are mean ± SEM. *P> 0.05, ** P> 0.01, and *** P> 0.001 at different time points compared to controls

Figure 2
Figure 2

Comparison of cytotoxicty effect of safranal on cell viability of prostate cancer (PC-3) and non-malignant (MRC-5) cell line after 48 h. Morphologic changes of cells after treatment with different concentrations of safranal. 1: MRC-5 cell line; 2: PC-3 cell line; a = 0; b = 10; c = 15, and d = 20 μg/ml safranal

Figure 3
Figure 3

DNA fragmentation of PC-3 exposed to safranal. Fragmentations of genomic DNA in PC-3 cells were treated with 20 μg/ml of safranal for 24, 48, and 72 h. DNA laddering formation was viewed on ethidium bromide-stained gel (2%) and photographed by UV illumination. M, molecular weight marker; C, DMSO control

Figure 4
Figure 4

Assessment of apoptosis by Annexin V/PI on human prostate cancer cells (PC-3). The cells were treated with 10 and 20 ƒÝg/ml safranal for 48 h (symbol II, III) or media only (control symbol I), and apoptosis was examined by flow cytometry after Annexin V/PI double staining. Necrotic cells lose membrane integrity, permitting PI entry. Viable cells exhibit Annexin V (−)/PI (−); early apoptotic cells exhibit Annexin (+)/PI (−); late apoptotic cells or necrotic cells exhibit Annexin V (+)/PI (+)

Figure 5
Figure 5

Assessment of apoptosis by Annexin V/PI on human prostate cancer cells (PC-3). Percentage of cell death based on the assessment of apoptosis by Annexin V/PI. *** indicates P> 0.001 compared to control

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