Effect of quercetin and chrysin and its association on viability and cell cycle progression in MDA-MB-231 and MCF-7 human breast cancer cells.

Pharmacological properties of flavonoids have been reported, with an anticancer role amongst them, however, its mechanisms are not fully elucidated. In this study, the activity of quercetin and chrysin towards MCF-7 and MDA-MB-231 breast cancer cells was investigated. Cellular viability was determined after treatment with the compounds in different concentrations for 24 h. Secondly, cells were treated with fixed concentration of chrysin and different concentrations of quercetin with preincubation for 1 h. Both compounds inhibited cellular proliferation in dose-dependent manner. The association showed improvement in their cytotoxicity, more expressively with preincubation of quercetin. Quercetin and chrysin association induced cell cycle arrest in sub-G0/G1 phase in MDA-MB-231 cells, modified the expression of caspases-3 and -8,-8, inducing late apoptosis cell death. Taken together, our results demonstrate that both flavonoids inhibited cells growth in a dose-dependent manner and the association of quercetin improved chrysin's toxic effect over the cell lines.

Oxyresveratrol in Breast Cancer Cells: Synergistic Effect with Chemotherapeutics Doxorubicin or Melphalan on Proliferation, Cell Cycle Arrest, and Cell Death.

Breast cancer is the second most common type of cancer in the world. Polyphenols can act at all stages of carcinogenesis and oxyresveratrol (OXY) promising anticancer properties, mainly associated with chemotherapy drugs. The aim of this study was to investigate the effect of OXY with doxorubicin (DOX) or melphalan (MEL), either isolated or associated, in MCF-7 and MDA-MB-231 breast cancer cells. Our results showed that OXY, DOX, and MEL presented cytotoxicity, in addition to altering cell morphology. The synergistic association of OXY + DOX and OXY + MEL reduced the cell viability in a dose-dependent manner. The OXY, DOX, or MEL and associations were able to alter the ROS production, ∆Ψm, and cell cycle; DOX and OXY + DOX led the cells to necrosis. Furthermore, OXY and OXY + MEL were able to lead the cells to apoptosis and upregulate caspases-3, -7, -8, and -9 in both cells. LC-HRMS showed that 7-deoxidoxorubicinone and doxorubicinol, responsible for the cardiotoxic effect, were not identified in cells treated with the OXY + DOX association. In summary, our results demonstrate for the first time the synergistic effect of OXY with chemotherapeutic agents in breast cancer cells, offering a new strategy for future animal studies.

Curcumin and melphalan cotreatment induces cell cycle arrest and apoptosis in MDA-MB-231 breast cancer cells.

Breast cancer is the second most common type of cancer worldwide and the leading cause of cancer death in women. Dietary bioactive compounds may act at different stages of carcinogenesis, including tumor initiation, promotion, and progression. Spices have been used for thousands of years and have many bioactive compounds with chemopreventive and chemotherapeutic properties. Curcumin has a multitude of beneficial biological properties, including anti-inflammatory and anticancer effects. This study investigated the effects of cotreatment with curcumin and the chemotherapeutic drug melphalan in cultured MDA-MB-231 breast cancer cells. When used alone, both curcumin and melphalan had a cytotoxic effect on breast cancer cells. Combined treatment with 11.65 µM of curcumin and 93.95 µM of melphalan (CURC/MEL) reduced cell viability by 28.64% and 72.43% after 24 h and 48 h, respectively. CURC/MEL reduced the number of colony-forming units and increased ROS levels by 1.36-fold. CURC/MEL alter cell cycle progression, induce apoptosis, and upregulate caspases-3, -7, and -9, in MDA-MB-231 cells. Cotreatment with curcumin and melphalan have anti-breast cancer cells effects and represent a promising candidate for clinical testing.

Low body mass index is associated with reduced intratumoral CD4+ T-lymphocyte infiltration in laryngeal squamous cell carcinoma patients.

Tumor size, inflammation, and nutritional status may be correlated with the immune response to cancer. Our hypothesis is that there is an interrelationship among tumor size, inflammatory response, and body mass index (BMI), and that these variables could alter T-lymphocyte infiltration in patients with laryngeal squamous cell carcinoma (LSCC). A retrospective cohort of 91 surgical LSCC patients treated at a Brazilian National Cancer Reference Center was followed for 5 years. We collected data regarding BMI, clinical factors, patients' lifestyle, C-reactive protein, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR). Data were obtained in the medical records within a maximum interval of 7 days before surgery. The stromal and intratumoral CD4+ and CD8+ T-cell infiltrations were obtained by immunohistochemistry. Our results demonstrated a significant correlation among tumor size and BMI, NLR, PLR, and LMR. Similarly, PLR and LMR were significantly correlated with BMI. Tumor size and inflammatory parameters were not associated with changes in T-cell infiltrations. However, patients with low BMIs had a significantly lower density of intratumoral CD4+ T lymphocytes infiltrated when compared with normal/high BMI patients (odds ratio, 0.14; 95% confidence interval, 0.03-0.58; P = .007). CD8+ T-lymphocyte infiltration did not change in low-BMI patients. In conclusion, we observed a correlation among tumor size, inflammation, and BMI. Tumor size/inflammation axis may be responsible for the change in BMI and, therefore, may have influenced the reduction of intratumoral CD4+ T-lymphocyte infiltration in LSCC patients.

Body Mass Index Alters the Predictive Value of the Neutrophil-to-Lymphocyte Ratio and Systemic Inflammation Response Index in Laryngeal Squamous Cell Carcinoma Patients.

Laryngeal squamous cell carcinoma (LSCC) is a frequent cancer subtype among head and neck cancers. Exacerbated inflammation and nutritional deficit are common features in this type of cancer and can be used as a prognostic marker. This study aimed to investigate the relationship between body mass index (BMI), neutrophil-to-lymphocyte ratio (NLR), and systemic inflammation response index (SIRI) on overall survival (OS) of LSCC patients. In this retrospective cohort study, 168 patients were followed for 5 years. Data on clinical factors, patients' life habits, height, weight, and hematological parameters were collected. BMI, NLR, and SIRI were calculated. Pretreatment NLR≥ 2.02 and SIRI≥ 1160.85 were independent prognostic factors for poor OS. Low BMI did not significantly affect the OS. However, the inflammatory parameters had their predictive capacity altered when stratified by the BMI classification. NLR≥ 2.02 + Low BMI or SIRI≥ 1160.85 + Low BMI increased in 8.6 and 3.8 times the risk of death, respectively. In contrast, stratification by normal/high BMI classification eliminated the predictive capacity of NLR and SIRI. Here, we demonstrated the possible ability of BMI to change the prognostic capacity of inflammatory markers NLR and SIRI in patients with LSCC.Supplemental data for this article is available online at https://doi.org/10.1080/01635581.2021.1952447.

Pro-Oxidant Effect of Resveratrol on Human Breast Cancer MCF-7 Cells is Associated with CK2 Inhibition.

Casein kinase 2 (CK2) plays a critical role in the proliferation and apoptosis of cancer cells. Resveratrol is a bioactive compound with anticancer and anti-inflammatory effects. This study investigated the pro-oxidant cytotoxic effects of resveratrol in association with the inhibition of CK2 activity on human breast carcinoma cells MCF-7. We showed that resveratrol and TBB, an inhibitor of CK2, decreased cell viability in a concentration dependent manner with an IC50 value of 238 µM and 106 µM after 24 h, of treatment, respectively. Resveratrol and TBB decreased CK2 activity by 1.6 and 1.4-fold, respectively, and both significantly decreased mitochondrial membrane potential. However, only resveratrol increased reactive oxygen species (ROS) levels by 1.7-fold as opposed to TBB, which did not affect ROS levels. Indeed, incubating MCF-7 cells with the antioxidant polyethylene glycol-catalase (PEG-CAT) preserved cell viability from the cytotoxic effects of resveratrol, but not from TBB toxicity. This effect seemed to be related to PEG-CAT ability to prevent CK2 inhibition induced by resveratrol incubation. In conclusion, this study demonstrated that the cytotoxic effect of resveratrol on MCF-7 cells might be associated with its pro-oxidant action, which inhibited CK2 activity, affecting cell viability and mitochondrial function.

Green Tea (Camellia sinensis) Extract Induces p53-Mediated Cytotoxicity and Inhibits Migration of Breast Cancer Cells.

Green tea (GT) has been shown to play an important role in cancer chemoprevention. However, the related molecular mechanisms need to be further explored, especially regarding the use of GT extract (GTE) from the food matrix. For this study, epigallocatechin gallate (EGCG) and epigallocatechin (EGC) were identified in GTE, representing 42 and 40% of the total polyphenols, respectively. MDA-MB-231 (p53-p.R280K mutant) and MCF-7 (wild-type p53) breast tumor cells and MCF-10A non-tumoral cells were exposed to GTE for 24-48 h and cell viability was assessed in the presence of p53 inhibitor pifithrin-α. GTE selectively targeted breast tumor cells without cytotoxic effect on non-tumoral cells and p53 inhibition led to an increase in viable cells, especially in MCF-7, suggesting the involvement of p53 in GTE-induced cytotoxicity. GTE was also effective in reducing MCF-7 and MDA-MD-231 cell migration by 30 and 50%, respectively. An increment in p53 and p21 expression stimulated by GTE was observed in MCF-7, and the opposite phenomenon was found in MDA-MB-231 cells, with a redistribution of mutant-p53 from the nucleus and no differences in p21 levels. All these findings provide insights into the action of GTE and support its anticarcinogenic potential on breast tumor cells.

Addition of Honey to an Apple and Passion Fruit Mixed Beverage Improves Its Phenolic Compound Profile.

The addition of honey to mixed beverages is interesting due to its contribution to the sweet taste, as well as because it is a dietary source of bioactive compounds. In this study, we investigated the chemical composition and sensory acceptance of an apple and passion fruit mixed beverage with added honey. The addition of honey did not produce a noticeable change in instrumental color but led to an increase in total soluble solids contents, and FRAP (20%), TEAC (72%), and DPPH (62%) values. The honey mixed beverages exhibited a better phenolic compound profile with an increase in catechin contents and an enrichment of quercetin when compared to the control mixed beverage, as well presenting good sensory acceptance. In conclusion, our results show that the addition of honey can be an alternative for improving the nutritional and sensorial characteristics of an apple and passion fruit mixed beverage.

Jaboticaba berry: A comprehensive review on its polyphenol composition, health effects, metabolism, and the development of food products.

Jaboticaba, a popular Brazilian berry, has been studied due to its relevant polyphenol composition, health benefits and potential use for the development of derived food products. Considering that around 200 articles have been published in recent years, this review aims to provide comprehensive and updated information, as well as a critical discussion on: (i) jaboticaba polyphenolic composition and extraction methods for their accurate determination; (ii) jaboticaba polyphenol's metabolism; (iii) biological effects of the fruit and the relationship with its polyphenols and their metabolites; (iv) challenges in the development of jaboticaba derived products. The determination of jaboticaba polyphenols should employ hydrolysis procedures during extraction, followed by liquid chromatographic analysis. Jaboticaba polyphenols, mainly anthocyanins and ellagitannins, are extensively metabolized, and their metabolites are probably the most important contributors to the relevant health effects associated with the fruit, such as antioxidant, anti-inflammatory, antidiabetic, hepatoprotective and hypolipidemic. Most of the technological processing of jaboticaba fruit and its residues is related to their application as a colorant, antioxidant, antimicrobial and source of polyphenols. The scientific literature still lacks studies on the metabolism and bioactivity of polyphenols from jaboticaba in humans, as well as the effect of technological processes on these issues.

Selective Cytotoxicity of Piperine Over Multidrug Resistance Leukemic Cells.

Multidrug resistance (MDR) is the main challenge in the treatment of chronic myeloid leukemia (CML), and P-glycoprotein (P-gp) overexpression is an important mechanism involved in this resistance process. However, some compounds can selectively affect MDR cells, inducing collateral sensitivity (CS), which may be dependent on P-gp. The aim of this study was to investigate the effect of piperine, a phytochemical from black pepper, on CS induction in CML MDR cells, and the mechanisms involved. The results indicate that piperine induced CS, being more cytotoxic to K562-derived MDR cells (Lucena-1 and FEPS) than to K562, the parental CML cell. CS was confirmed by analysis of cell metabolic activity and viability, cell morphology and apoptosis. P-gp was partially required for CS induction. To investigate a P-gp independent mechanism, we analyzed the possibility that poly (ADP-ribose) polymerase-1 (PARP-1) could be involved in piperine cytotoxic effects. It was previously shown that only MDR FEPS cells present a high level of 24 kDa fragment of PARP-1, which could protect these cells against cell death. In the present study, piperine was able to decrease the 24 kDa fragment of PARP-1 in MDR FEPS cells. We conclude that piperine targets selectively MDR cells, inducing CS, through a mechanism that might be dependent or not on P-gp.

Profile of polyphenol intake by women with different classes of obesity: Consumption of these compounds does not reflect healthy eating.

OBJECTIVES: The aim of this study was to evaluate polyphenol intake in women with different classes of obesity and identify which are consumed more frequently and what the food sources are. METHODS: A cross-sectional study was conducted with 114 women with obesity. The study evaluated polyphenol intake via a 3-d food record using Phenol-Explorer. Anthropometric, biochemical, and dietetic variables were evaluated. RESULTS: The women's habitual food intake was low calorie and adequate in macronutrients. Mean polyphenol intake by the group was 573 ± 490, 614 ± 475, and 379 ± 25 mg/d for class I, class II, and class III obesity (P = 0.002), respectively. The most frequent food or beverage consumed by the group was coffee and caffeoylquinic acid, its phenolic compound. Fruits, vegetables, and nuts contributed the least to the intake of polyphenols. CONCLUSIONS: Although the diets of the study participants did include some food sources of polyphenols, they were not of sufficient quality to significantly contribute to a healthy diet; instead, they sometimes were foods may have that contributed to weight gain. Women with class III obesity consumed the most calories; however, they had low fruit, vegetable, and whole foods intake.

Anti-Leishmania amazonensis activity of the terpenoid fraction from Eugenia pruniformis leaves.

Leishmaniasis is caused by protozoan parasites belonging to the genus Leishmania and includes cutaneous, mucocutaneous and visceral clinical forms. Drugs currently available for leishmaniasis treatment present high toxicity, and development of parasite resistance. Plants constitute an important source of compounds with leishmanicidal potential. This study aimed to evaluate the anti-Leishmania amazonensis activity of the terpenoid fraction of Eugenia pruniformis leaves (TF-EpL). TF-EpL was active against the promastigote and intracellular amastigote forms of L. amazonensis with IC50(24h) value of 43.60µg/mL and 44.77µg/mL, respectively. TF-EpL altered the cell cycle of the parasite, increasing 2.32-fold the cells in the Sub-G0/G1 phase. TF-EpL also changed the ΔΨm and increased ROS and the number of annexin-V-PI positive promastigotes, which suggests incidental death. ß-sitosterol, ursolic acid, corosolic acid and asiatic acid were isolated from TF-EpL. The results showed the antileishmanial activity of E. pruniformis terpenoids and its potential for further studies as a source of new drugs for leishmaniasis.

Bioactive Compounds and Metabolites from Grapes and Red Wine in Breast Cancer Chemoprevention and Therapy.

Phytochemicals and their metabolites are not considered essential nutrients in humans, although an increasing number of well-conducted studies are linking their higher intake with a lower incidence of non-communicable diseases, including cancer. This review summarizes the current findings concerning the molecular mechanisms of bioactive compounds from grapes and red wine and their metabolites on breast cancer-the most commonly occurring cancer in women-chemoprevention and treatment. Flavonoid compounds like flavonols, monomeric catechins, proanthocyanidins, anthocyanins, anthocyanidins and non-flavonoid phenolic compounds, such as resveratrol, as well as their metabolites, are discussed with respect to structure and metabolism/bioavailability. In addition, a broad discussion regarding in vitro, in vivo and clinical trials about the chemoprevention and therapy using these molecules is presented.

Resveratrol, Curcumin and Piperine Alter Human Glyoxalase 1 in MCF-7 Breast Cancer Cells.

Breast cancer is the leading cause of cancer mortality in women worldwide. Conventional cancer treatment is costly and results in many side effects. Dietary bioactive compounds may be a potential source for breast cancer prevention and treatment. In this scenario, the aim of this study was to investigate the effects of the bioactive compounds resveratrol, curcumin and piperine (R-C-P) on MCF-7 breast cancer cells and to associate them to Glyoxalase 1 (GLO1) activity. The findings indicate that R-C-P exhibits cytotoxicity towards MCF-7 cells. R-C-P decreased mitochondrial membrane potential (ΔΨm) by 1.93-, 2.04- and 1.17-fold, respectively. Glutathione and N-acetylcysteine were able to reverse the cytotoxicity of the assessed bioactive compounds in MCF-7 cells. R-C-P reduced GLO1 activity by 1.36-, 1.92- and 1.31-fold, respectively. R-C-P in the presence of antimycin A led to 1.98-, 1.65- and 2.16-fold decreases in D-lactate levels after 2 h of treatment, respectively. Glyoxal and methylglyoxal presented cytotoxic effects on MCF-7 cells, with IC50 values of 2.8 and 2.7 mM and of 1.5 and 1.4 mM after 24 and 48 h of treatment, respectively. In conclusion, this study demonstrated that R-C-P results in cytotoxic effects in MCF-7 cells and that this outcome is associated with decreasing GLO1 activity and mitochondrial dysfunction.

Resveratrol Modifies Lipid Composition of Two Cancer Cell Lines.

Resveratrol (Resv) offers health benefits in cancer and has been reported to modulate important enzymes of lipid metabolism. Studies of its effects on lipid composition in different subtypes of breast-cancer cells are scarce. Thus, we investigated the alterations in phospholipids (PL), fatty acids (FA), and lipid metabolism enzymes in two breast-cancer cell lines after Resv treatment. MCF-7 and MDA-MB-231 cells were treated with 80 and 200 µM of Resv, respectively, for 24 hours. We analyzed PL with radiolabeled inorganic phosphate (32Pi) by thin-layer chromatography, FA by gas chromatography-mass spectrometry, and lipid metabolism enzymes (DGAT2, FAS, ρACCß, pAMPKα, and AMPK) by Western blot. Resv treated MDA-MB-231 phospholipids showed a reduction in phosphatidylcholine (63%) and phosphatidylethanolamine (35%). We observed an increase in eicosapentaenoic acid (EPA) (73%) and docosahexaenoic acid (DHA) (65%) in MCF-7 cells after Resv treatment. Interestingly, the same treatment caused 50% and 90% increases in EPA and DHA, respectively, in MDA-MB-231 cells. In MCF-7 cells, Resv increased the expression of ρACCß (3.3-fold) and AMPKα/ρAMPKα (1.5-fold) and in MDA-MB-231 cells it inhibited the expression of ρACCß (111.8-fold) and AMPKα/ρAMPKα (1.2 fold). Our results show that Resv modified PL and saturated and unsaturated FA especially in MDA-MB-231 cells, and open new perspectives to the understanding of the reported anticancer effect of Resv on these cells.

Increase in fatty acids and flotillins upon resveratrol treatment of human breast cancer cells.

Flotillin-1 and flotillin-2 are highly conserved proteins that localize into cholesterol-rich microdomains in cellular membranes. Flotillins are closely related to the occurrence and development of various types of human cancers. Flotillin-1 is highly expressed in breast cancer, and the high expression level of flotillin-1 is significantly correlated with poorer patient survival. Here we studied the relationship between the formation of lipid rafts and the expression of flotillins and lipids in human breast cancer cells. We used the polyphenol compound resveratrol to alter the structure and function of the plasma membrane. Our data revealed an increase in fatty acids in MCF-7 and MDA-MB-231 cells upon resveratrol treatment. Interestingly, we also found an increase in the expression of both flotillin-1 and flotillin-2 in breast tumor cells after treatment. Resveratrol also induced changes in the pattern of flotillin distribution among detergent-resistant lipid rafts fractions in both cell lines and induced the nuclear translocation of flotillin-2. Since resveratrol has been pointed out as a putative cancer therapy agent, our results could have an impact on the understanding of the effects of resveratrol in tumor cells.

Cancer Chemoprevention by Resveratrol: The p53 Tumor Suppressor Protein as a Promising Molecular Target.

Increasing epidemiological and experimental evidence has demonstrated an inverse relationship between the consumption of plant foods and the incidence of chronic diseases, including cancer. Microcomponents that are naturally present in such foods, especially polyphenols, are responsible for the benefits to human health. Resveratrol is a diet-derived cancer chemopreventive agent with high therapeutic potential, as demonstrated by different authors. The aim of this review is to collect and present recent evidence from the literature regarding resveratrol and its effects on cancer prevention, molecular signaling (especially regarding the involvement of p53 protein), and therapeutic perspectives with an emphasis on clinical trial results to date.

Four conventional soybean [Glycine max (L.) Merrill] seeds exhibit different protein profiles as revealed by proteomic analysis.

Soybeans have several functional properties due to their composition and may exert beneficial health effects that are attributed to proteins and their derivative peptides. The present study aimed to analyze the protein profiles of four new conventional soybean seeds (BRS 257, BRS 258, BRS 267, and Embrapa 48) with the use of proteomic tools. Two-dimensional (2D) and one-dimensional (1D) gel electrophoreses were performed, followed by MALDI-TOF/TOF and ESI-Q-TOF mass spectrometry analyses, respectively. These two different experimental approaches allowed the identification of 117 proteins from 1D gels and 46 differentially expressed protein spots in 2D gels. BRS 267 showed the greatest diversity of identified spots in the 2D gel analyses. In the 1D gels, the major groups were storage (25-40%) and lipid metabolism (11-25%) proteins. The differences in protein composition between cultivars could indicate functional and nutritional differences and could direct the development of new cultivars.

Transient transfection of a wild-type p53 gene triggers resveratrol-induced apoptosis in cancer cells.

Resveratrol is a promising chemopreventive agent that mediates many cellular targets involved in cancer signaling pathways. p53 has been suggested to play a role in the anticancer properties of resveratrol. We investigated resveratrol-induced cytotoxicity in H1299 cells, which are non-small lung cancer cells that have a partial deletion of the gene that encodes the p53 protein. The results for H1299 cells were compared with those for three cell lines that constitutively express wild-type p53: breast cancer MCF-7, adenocarcinomic alveolar basal epithelia A549 and non-small lung cancer H460. Cell viability assays revealed that resveratrol reduced the viability of all four of these cell lines in a dose- and time-dependent manner. MCF-7, A549 and H460 cells were more sensitive to resveratrol than were H1299 cells when exposed to the drug for 24 h at concentrations above 100 µM. Resveratrol also increased the p53 protein levels in MCF-7 cells without altering the p53 mRNA levels, suggesting a post-translational modulation of the protein. The resveratrol-induced cytotoxicity in these cells was partially mediated by p53 and involved the activation of caspases 9 and 7 and the cleavage of PARP. In H1299 cells, resveratrol-induced cytotoxicity was less pronounced and (in contrast to MCF-7 cells) cell death was not accompanied by caspase activation. These findings are consistent with the observation that MCF-7 cells were positively labeled by TUNEL following exposure to 100 µM resveratrol whereas H1299 cells under similar conditions were not labeled by TUNEL. The transient transfection of a wild-type p53-GFP gene caused H1299 cells to become more responsive to the pro-apoptotic properties of resveratrol, similarly to findings in the p53-positive MCF-7 cells. Our results suggest a possible therapeutic strategy based on the use of resveratrol for the treatment of tumors that are typically unresponsive to conventional therapies because of the loss of normal p53 function.

Cellular antioxidant activity of feijoada whole meal coupled with an in vitro digestion.

Consumption of plant food rich meals, such as feijoada, a traditional meal in Brazil, is associated with the reduction of chronic disease. The objectives of this study were to determine phytochemical content and antioxidant activity by chemical and cellular antioxidant assays (CAA) of feijoada with or without in vitro digestion. Feijoada showed no difference in phenolics and flavonoids after digestion. Bound and residue contributions to total phenolics were 20.9% and 32.2%, respectively, suggesting that phenolics reach the colon after intake. Flavonoids in residue and bound fractions represented 50% of total flavonoids. Antioxidant activity of feijoada without digestion was higher than that with digestion; however, it showed lower antiproliferative activity and CAA. Feijoada with in vitro digestion also yielded phenolics with higher CAA. Analyses of whole meals should be used to evaluate phytochemicals present in food mixtures consumed, especially with digestion models coupled with CAA resulting in information similar to those in physiological conditions.