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Comparison of Three Different Methods for Determining Cell Proliferation in Breast Cancer Cell Lines
Published on: September 3, 2016
Different propolis samples, phenolic content, and breast cancer cell lines: Variable cytotoxicity ranging from
Mehmet Fatih Seyhan1, Eren Yılmaz1, Özlem Timirci-Kahraman1
1Department of Molecular Medicine, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey.
Abstract:
Researchers have started focusing on investigating the anticarcinogenic effects of natural products with the slightest side effects possible, because current breast cancer treatment approaches are unable to achieve absolute success especially on aggressive subtypes. Propolis is among these products with its antimicrobial, antifungal, anti-inflammatory, and anticancer effects. Therefore, seven different samples were collected from different regions (Argentina, China, and Istanbul-Turkey) and applied on nonaggressive breast cancer cell line (BCCL) MCF-7 and aggressive cell lines SK-BR-3, and MDA-MB-231. Initially, the phenolic/flavonoid constituents of the propolis ethanol extracts were investigated by liquid chromatography-mass spectrometry-mass spectrometry (LS-MS/MS) and high-performance liquid chromatography (HPLC) analyses. Then, the anticarcinogenic effects of the propolis samples on MCF-7, SK-BR-3, MDA-MB-231 were evaluated by WST1 analysis and only selected ones on MCF-10A and hPdLF. According to the LS-MS/MS and HPLC analysis, Turkey originated propolis (Turkey3) were found to be richer than the other propolis samples in terms of phenolic/flavonoid compounds. Turkey propolis significantly inhibited cell proliferation in both nonaggressive and aggressive BCCL (P < 0.01). Therefore, Turkey3 propolis was selected for further evaluation using Annexin V-PI apoptosis detection assays. In addition, selected compounds among the propolis contents such as galangin, caffeic acid, apigenin, quercetin, and ferulic acid were applied to the MCF-7 cell line to detect cytotoxic and apoptotic effects. Galangin, caffeic acid, apigenin, and quercetin remarkably induced cell proliferation inhibition at all time intervals, whereas ferulic acid was found non efficient on the MCF-7 cell line. Annexin V-PI assay clarified that all cell proliferation inhibitions were markedly apoptotic. Our findings indicated that the inhibition effect of propolis on breast cancer cell proliferation was in a propolis type-, dose- and time-dependent fashion. Turkey3 propolis showed statistically significant cytotoxic effects on both the nonaggressive and aggressive BCCL. These findings were consistent with the effects of its rich phenolic and flavonoid contents, in terms of variety. © 2018 IUBMB Life, 71(5):619-631, 2019.
Insights
This study shows Turkish propolis, rich in phenolic compounds, effectively inhibits breast cancer cell growth through apoptosis. It offers a promising natural approach for breast cancer treatment, especially for aggressive subtypes.
Area of Science:
- Natural Products Chemistry
- Cancer Biology
- Pharmacology
Background:
- Current breast cancer treatments have limitations, particularly for aggressive subtypes.
- Natural products are being investigated for their potential anticarcinogenic effects with fewer side effects.
- Propolis, known for its antimicrobial and anti-inflammatory properties, is a candidate for cancer therapy.
Purpose of the Study:
- To evaluate the anticarcinogenic effects of propolis from different regions on breast cancer cell lines.
- To identify the phenolic and flavonoid compounds responsible for propolis's anticancer activity.
- To determine the mechanism of action, specifically focusing on apoptosis induction.
Main Methods:
- Propolis samples were analyzed for phenolic and flavonoid content using Liquid Chromatography-Mass Spectrometry-Mass Spectrometry (LS-MS/MS) and High-Performance Liquid Chromatography (HPLC).
- Anticancer effects were assessed using WST1 assays on nonaggressive (MCF-7) and aggressive (SK-BR-3, MDA-MB-231) breast cancer cell lines.
- Apoptosis was confirmed using Annexin V-PI assays, and specific compounds (galangin, caffeic acid, apigenin, quercetin, ferulic acid) were tested individually.
Main Results:
- Turkish propolis (Turkey3) exhibited a higher concentration of phenolic and flavonoid compounds compared to other samples.
- Turkey3 propolis significantly inhibited proliferation in both nonaggressive and aggressive breast cancer cell lines (P < 0.01).
- Specific compounds like galangin, caffeic acid, apigenin, and quercetin demonstrated significant cytotoxic and apoptotic effects, while ferulic acid was less effective.
Conclusions:
- Propolis exhibits dose- and time-dependent inhibition of breast cancer cell proliferation.
- The observed anticancer effects are attributed to the rich variety of phenolic and flavonoid constituents in propolis.
- Turkish propolis demonstrates significant cytotoxic and apoptotic effects, highlighting its potential as a natural therapeutic agent for breast cancer.
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