Viability Of MCF-7 Breast Cancer Cells Research Articles

Effects of matcha tea extract on cell viability and estrogen receptor-β expression on MCF-7 breast cancer cells

PurposeIn the following work, we investigated the effect of matcha green tea extract (MTE) on MCF-7 breast cancer cell viability and estrogen receptor-beta expression (ERβ).MethodsMCF-7 cells were stimulated with MTE at concentrations of 5 and 10 µg/ml. Cell viability was assessed using a water-soluble tetrazolium assay (WST-1 assay) after an incubation time of 72 h. ERβ was quantified at gene level by real-time polymerase chain reaction (PCR). A western blot (WB) was carried out for the qualitative assessment of the expression behavior of on a protein level.ResultsThe WST-1 test showed a significant inhibition of viability in MFC-7 cells after 72 h at 10 µg/ml. The WB demonstrated a significant quantitative decrease of ERβ at protein level with MTE concentrations of 10 µg/ml. In contrast, the PCR did not result in significant downregulation of ERβ.ConclusionMTE decreases the cell viability of MCF-7 cells and furthermore leads to a decrease of ERβ at protein level.

Genistein and coumestrol reduce MCF-7 breast cancer cell viability and inhibit markers of preferential metastasis, bone matrix attachment and tumor-induced osteoclastogenesis

The propensity of breast cancer to preferentially metastasize to the skeleton is well known. Once established in bone metastatic breast cancers have a poor prognosis due to their ability to promote extensive bone loss which augments tumor burden. Unfortunately, current anti-resorptive therapies for skeletal metastasis are typically prescribed after secondary tumors have formed and are palliative in nature. One group of compounds with the potential to reduce both tumor burden and osteolysis are phytoestrogens (PE), but the mechanisms mediating a beneficial effect are unclear. Therefore, the current study examined the effect of genistein and coumestrol alone or in combination on breast cancer cell number, expression of mediators of preferential skeletal metastasis, bone matrix attachment and tumor-induced osteoclast formation. Results showed that genistein and coumestrol significantly reduced viable cell number in an estrogen receptor dependent manner (p < 0.05), whereas combinations of PE had no effect. In addition, genistein and coumestrol significantly reduced expression of genes driving epithelial to mesenchymal transition (snail), bone attachment (CXCR4 and integrin αV) and osteolysis (PTHrP and TNF-α). In keeping with this genistein and coumestrol significantly suppressed attachment of breast cancer cells to bone matrix and inhibited tumor and RANKL-induced osteoclast formation. Our data suggests that phytoestrogens not only decrease breast cancer cell viability but also antagonize essential tumor bone interactions that establish and drive the progression of skeletal metastasis.

Bioconversion of Genistein to Orobol by Bacillus subtilis Spore Displayed Tyrosinase and Monitoring the Anticancer Effects of Orobol on MCF-7 Breast Cancer Cells

Orobol (5,7,3′,4′-tetrahydroxyisoflavone) is a highly hydroxylated isoflavone, which is rarely found in natural environment. In this study, orobol was produced due to bioconversion of a soybean frequented isoflavone, genistein (4′,5,7-Trihydroxyisoflavone) by an active, stable, reusable genetically immobilized enzyme, the recombinant Bacillus subtilis spore displayed tyrosinase. Thin layer chromatography and high performance liquid chromatography were used to monitor the reaction. The results revealed that 1 mM orobol was produced from 3′-hydroxylation of 1 mM genistein by spore displayed tyrosinase at 37°C during 90 min incubation. To study on anti-proliferative effects of orobol, MCF-7 breast cancer cell viability was determined by MTT method and flow cytometric analysis. The comparison between reduction in cell viabilities in 50 to 500 µM genistein and orobol treated cells revealed that orobol has more remarkable anticancer effects than genistein. Flow cytometric analysis showed more than 87% cytotoxicity in 500 µM orobol treated cells by flow cytometric analysis. To the best of our knowledge this is the first report of the orobol demonstrated potent anticancer activity against MCF-7 breast cancer cell. It is suggested that enzymatic biotransformation of soybean genistein to orobol will be made a new approach to create highly bioactive products usable in food and pharmaceutical industries.

Cytotoxicity of Caffeine on MCF-7 Cells Measured by XTT Cell Proliferation Assay (P06-038-19)

ObjectivesCaffeine is a widely consumed phytochemical naturally occurring in coffee and tea, or added to beverages such as energy drinks used to promote wakefulness and alertness. Previous research suggests that MCF-7 breast cancer cell viability decreases after exposure to millimolar (mM) amounts of caffeine in vitro. The effect of smaller, micromolar (μM), amounts of caffeine has not been fully investigated. This in vitro study investigates the impact of varying μM concentrations of caffeine on MCF-7 cell proliferation. MethodsMCF-7 cells were cultured and treated with caffeine concentrations that followed a serial dilution curve from 10 μM to 1.2 mM, and 5 mM of caffeine, a concentration that previous study has suggested as cytotoxic to MCF-7 cells. Both a media control (0 μM caffeine concentration) and a vehicle control (distilled water) were included. After 24 hours of treatment exposure, an XTT assay calculated MCF-7 cell proliferation. The experiment was run in triplicate and each caffeine concentration was tested three times during each run. ResultsAfter a 24-hour period, a bimodal result was noted. When compared to the media control (0 μM caffeine concentration), the 10 μM caffeine concentration showed 91–107% cell proliferation; 20 μM at 104–145%; 40 μM at 99–115%; 80 μM at 80–96%; 160 μM at 80–83%; 320 μM at 76–97%; 640 μM at 65–81%; 1.2 mM at 57–80%; 5 mM at 37–50%. A paired samples t-test done for the vehicle control indicated effects on cell proliferation were due to the caffeine concentration, rather than the distilled water (p = 0.54). ConclusionsThis data reflects previous research suggesting that caffeine is cytotoxic to MCF-7 cells in vitro at the 5 mM concentration. In addition, percentage and t-test data suggest μM concentrations of caffeine have a bimodal effect on MCF-7 cells in vitro, with an initial increase in cell proliferation from 10 μM to 40 μM of caffeine and a potential starting concentration of cytotoxic effect at 80 μM of caffeine. Funding SourcesFunding provided by Bastyr University Faculty Student Research Grant.

PIPERINE IN THE PREVENTION OF THE DECREASED TAMOXIFEN SENSITIVITY IN MCF-7 BREAST CANCER CELL LINE

Objective: This study was designed to analyze the role of piperine in modulating P-glycoprotein mRNA expression when added in combination withtamoxifen to breast cancer cells in culture.Methods: MCF-7 breast cancer cells were treated with 1 μM tamoxifen with or without piperine (12.5, 25, or 50 μM) or verapamil 50 μM (P-glycoproteininhibitor positive control) for up to 12 days. We assessed the cell viability and isolated total RNA from them. We quantified P-glycoprotein expressionsusing quantitative reverse transcription polymerase chain reaction.Results: Administration of various doses of piperine decreased MCF-7 breast cancer cell viability. Piperine, when given in combination with tamoxifen,decreased the expression of P-glycoprotein mRNA in cells compared with the expression in cells treated with tamoxifen only. The effects were shownto be dose dependent.Conclusion: Piperine prevents the development of breast cancer cell tamoxifen resistance, probably through its inhibition of P-glycoprotein expression.

A Combination of Curcumin from Turmeric and Alpha-linolenic Acid Shows Antagonism with MCF-7 Breast Cancer Cells in Phenol-red Free Medium

Aims: To determine the total phenols content and antioxidant capacity for turmeric and curcumin, and to assess the effect of alpha-Linolenic acid (ALA) combinations treatments on MCF-7 breast cancer cell viability and intracellular reactive oxygen species (ROS).Study Design: In-vitro study.Place and Duration of Study: School of Biomedical Sciences, Ulster University, Coleraine (UK) September 2015 to September 2016.Methods: Curcumin was characterized for total phenols content (TPC) and antioxidant capacity (AOC) using Folin-Denis and ABTS (2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid diammonium salt) assays. MCF-7 cells were grown in DMEM phenol-red free medium (+ 10% charcoal stripped foetal bovine serum) and treated with curcumin, ALA or their combinations. Cytotoxicity was assessed using the sulforhodamine-B assay. Intracellular ROS was monitored using 2,7-dichlorodihydroflourescein diacetate assay.Results: Curcumin showed 42-50 folds higher TPC and AOC compared to turmeric. Both curcumin and ALA (0-500 µM) inhibited MCF-7 cells with the 50% effective dose (EC50) equal to 32 µM (curcumin) or 117 µM (ALA). Combination of curcumin and ALA led to EC50 values of 221 µM (curcumin) and 304 µM (ALA). Isobologram analysis and values for Combination index (CI; CI>1.0) are consistent with ALA and curcumin antagonism. Changes of intracellular ROS were 20-fold higher with ALA treatment of MCF-7 compared with curcumin.Conclusions: ALA and curcumin were each cytotoxic towards MCF-7 breast cancer cells but their combination decreases the effectiveness of each agent due to antagonistic interactions. Both ALA and curcumin produce rises in intracellular ROS for MCF-7 cells. The wider implications of such findings is that though dietary antioxidants could be beneficial on their own, antagonistic interaction with ALA, n-3 fatty acids and other ROS generating conventional anti-cancer drug could be of concern.

Curcumin enhanced antiproliferative effect of mitomycin C in human breast cancer MCF-7 cells in vitro and in vivo

To investigate the efficacy of mitomycin C (MMC) in combination with curcumin in suppressing human breast cancer in vitro and in vivo. Human breast cancer MCF-7 cells were used. Cell viability was measured using MTT assay. The cell cycle phase was detected with flow cytometric analysis. Cell cycle-associated proteins were examined using Western blot analysis. MCF-7 breast cancer xenografts were established to monitor tumor growth and cell cycle-associated protein expression. Curcumin inhibited MCF-7 breast cancer cell viability in a concentration-dependent manner (IC(50) value=40 μmol/L). Similarly, MMC inhibited the cell viability with an IC(50) value of 5 μmol/L. Combined treatment of MMC and curcumin showed a synergistic antiproliferative effect. In the presence of curcumin (40 μmol/L), the IC(50) value of MMC was reduced to 5 μmol/L. In MCF-7 xenografts, combined administration of curcumin (100 mg/kg) and MMC (1-2 mg/kg) for 4 weeks produced significantly greater inhibition on tumor growth than either treatment alone. The combined treatment resulted in significantly greater G(1) arrest than MMC or curcumin alone. Moreover, the cell cycle arrest was associated with inhibition of cyclin D1, cyclin E, cyclin A, cyclin-dependent kinase 2 (CDK2) and CDK4, along with the induction of the cell cycle inhibitor p21 and p27 both in MCF-7 cells and in MCF-7 xenografts. These proteins were regulated through p38 MAPK pathway. The results suggest that the combination of MMC and curcumin inhibits MCF-7 cell proliferation and cell cycle progression in vitro and in vivo via the p38 MAPK pathway.