Enterolactone reduces telomerase activity and the level of its catalytic subunit in breast cancer cells

Objective: There is a positive correlation between higher serum phytoestrogen concentrations and lower risk of breast cancer. The activation of telomerase is crucial for the growth of cancer cells; therefore, the aim of this study was to examine the effects of enterolactone [ENL] and enterodiol [END] on this enzyme

Materials and Methods: In this experimental study, we performed the viability assay to determine the effects of different concentrations of ENL and END on cell viability, and the effective concentrations of these two compounds on cell growth. We used western blot analysis to evaluate human telomerase reverse transcriptase catalytic subunit [hTERT] expression and polymerase chain reaction [PCR]-ELISA based on the telomeric repeat amplification protocol [TRAP] assay for telomerase activity

Results: Both ENL and END, at 100 micro M concentrations, significantly [P<0.05] reduced cell viability. However, only the 100 micro M concentration of ENL significantly [P<0.05] decreased hTERT protein levels and telomerase activity. Lower concentrations of ENL did not have any significant effects on telomerase activity and hTERT protein levels

Conclusion: High concentration of ENL decreased the viability of MCF-7 breast cancer cells and inhibited the expression and activity of telomerase in these cells. Although END could reduce breast cancer cell viability, it did not have any effect on telomerase expression and activity

Inhibition of Nicotinamide Phosphoribosyltransferase Induces Apoptosis in Estrogen Receptor-Positive MCF-7 Breast Cancer Cells / 한국유방암학회지

PURPOSE: Tumor cells have increased turnover of nicotinamide adenine dinucleotide (NAD⁺), the main coenzyme in processes including adenosine diphosphate-ribosylation, deacetylation, and calcium mobilization. NAD⁺ is predominantly synthesized in human cells via the salvage pathway, with the first component being nicotinamide. Nicotinamide phosphoribosyltransferase (NAMPT) is the key enzyme in this pathway, and its chemical inhibition by FK866 has elicited antitumor effects in several preclinical models of solid and hematologic cancers. However, its efficacy in estrogen receptor (ER)-positive and human epidermal growth factor receptor 2-positive breast cancer cells has not been previously investigated. In this study, we aimed to deplete the NAD⁺ content of MCF-7 cells, a model cell line for ER-positive breast cancer, by inhibiting NAMPT in order to evaluate downstream effects on p53 and its acetylation, p21 and Bcl-2-associated X protein (BAX) expression, and finally, apoptosis in MCF-7 breast cancer cells. METHODS: MCF-7 cells were cultured and treated with FK866. NAD⁺ levels in cells were determined colorimetrically. Levels of p53 and its acetylated form were determined by Western blotting. Expression of p21 and BAX was determined by real-time polymerase chain reaction. Finally, levels of apoptosis were assessed by flow cytometry using markers for annexin V and propidium iodide. RESULTS: FK866 treatment was able to increase p53 levels and acetylation, upregulate BAX and p21 expression, and induce apoptosis in MCF-7 cells. Addition of exogenous NAD⁺ to cells reversed these effects, suggesting that FK866 exerted its effects by depleting NAD⁺ levels. CONCLUSION: Results showed that FK866 could effectively inhibit NAD⁺ biosynthesis and induce programmed cell death in MCF-7 cells, suggesting that NAMPT inhibitors may be useful for the treatment of ER-positive breast cancers.

17 beta-estradiol stimulates generation of reactive species oxygen and nitric oxide in ovarian adenocarcinoma cells [OVCAR 3]

Experimental and epidemiological evidence supports a role for steroid hormones in the pathogenesis of ovarian cancer. Among steroid hormones, 17 beta -estradiol [E2] has the most potent effect on proliferation, apoptosis and metastasis. In the present study, we investigated the effect of E2 on production of ROS and NO in ovarian cancer cells. Ovarian adenocarcinoma cell line [OVCAR-3] was cultured and treated with various concentrations of E2, antioxidants [N-acetyle cysteine and Ebselen] and ICI182780 as an estrogen receptor antagonist. MTT test was performed to evaluate cell viability. NO and ROS levels were measured by Griess and DCFH-DA methods, respectively. ROS levels as well as NO levels were increased in OVCAR-3 cells treated with E2. The increase in ROS production was in parallel with increased cell viability which indicates that estrogen-induced ROS can participate in cancer progression. ICI182780 abolished E2-induced ROS production. Progesterone was also effective in reducing ROS and NO generation. NO and ROS are important molecules in signaling networks in cell. These molecules can be used as therapeutic targets for prevention and treatment of ovary cancer and other estrogen-induced malignancies