Studying the anticancer properties of Parthenolide (PTL) in MCF-7 breast cancer cells

Date

2018-08

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Abstract

Background: Breast cancer is considered as the most prevalent cancer in women world-wide. The average risk for a woman developing breast cancer in her life is about 12% in United States. Cancer cells including breast cancer are resistant to apoptosis and undergo epithelial to mesenchymal transition (EMT) for growth and proliferation. Thus, induction of apoptosis and reversal of EMT process might be effective to prevent cancer cell proliferation and tumor growth. Parthenolide (PTL) is a multifunctional naturally occurring compound. It is considered as a novel anti-tumor agent. Parthenolide induces cytotoxicity in several cancers. However, its anticancer properties in breast cancer cells are not completely known. In this study, low doses of PTL were used to determine its effects on cellular properties, EMT, and angiogenesis in MCF-7 cells and also exploredits underlying molecular mechanism.

Methods and materials: MCF-7 breast cancer cells were obtained from American Type Culture Collection (ATCC). Cells were grown in DMEM containing 10% fetal bovine serum and 1% penicillin streptomycin, at 37°C in 5% CO2 incubator. For treating the cells, Parthenolide (PTL) 2 and 5 μM was used for 24 hours using DMEM media with 0.5% FBS. Cell viability was measured using the MTT (3-(4,5-dimethy lthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay kit (BioVision k299-1000). Monolayer wound scratch assay was done for cell migration analysis by using a Nikon Eclipse motorized microscope with incubator (Nikon Instruments Inc.) at 5% CO2 and 37ºC temperature. To detect apoptosis, Apoptosis Detection Kit I (BD Biosciences) was used and the results were analyzed by using a Nikon Eclipse motorized microscope with incubator (Nikon Instruments Inc.). Cytosolic and nuclear extracts were prepared and subjected to Western blot analysis. Proteins and genes implicated in autophagy, EMT and angiogenesis are analyzed by western blot and quantitative polymerase chain reaction (qPCR). For statistical analysis, one-way ANOVA was used. All values are reported as mean ± standard error. Differences were considered to be statistically significant at P values 0.05 or less.

Results and Conclusion: Low doses of (2 & 5 µM) of PTL decreases viability, inhibits cell migration, induces apoptosis, and increases autophagy in MCF-7 cells. Interestingly, PTL treatment reverses EMT in MCF7 cells. In addition, PTL activates AMPK as evaluated by identifying phosphorylated form of AMPK. Though we conducted our study in only MCF-7 cells, our results suggest that PTL might be an effective therapeutic option against breast cancer.

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Keywords

Breast cancer, Parthenolide, Apoptosis, EMT, Angiogenesis

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