The Molecular Basis of Different Approaches for the Study of Cancer Stem Cells and the Advantages and Disadvantages of a Three-Dimensional Culture.

Cancer stem cells (CSCs) are a rare tumor subpopulation with high differentiation, proliferative and tumorigenic potential compared to the remaining tumor population. CSCs were first discovered by Bonnet and Dick in 1997 in acute myeloid leukemia. The identification and isolation of these cells in this pioneering study were carried out through the flow cytometry, exploiting the presence of specific cell surface molecular markers (CD34+/CD38-). In the following years, different strategies and projects have been developed for the study of CSCs, which are basically divided into surface markers assays and functional assays; some of these techniques also allow working with a cellular model that better mimics the tumor architecture. The purpose of this mini review is to summarize and briefly describe all the current methods used for the identification, isolation and enrichment of CSCs, describing, where possible, the molecular basis, the advantages and disadvantages of each technique with a particular focus on those that offer a three-dimensional culture.

Nutritional Value and Preventive Role of Nigella sativa L. and Its Main Component Thymoquinone in Cancer: An Evidenced-Based Review of Preclinical and Clinical Studies.

In recent times, scientific attention has been paid to different foods and their bioactive components for the ability to inhibit the onset and progress of different types of cancer. Nigella sativa extract, powder and seed oil and its main components, thymoquinone and α-hederin, have showed potent anticancer and chemosensitizing effects against various types of cancer, such as liver, colon, breast, renal, cervical, lung, ovarian, pancreatic, prostate and skin tumors, through the modulation of various molecular signaling pathways. Herein, the purpose of this review was to highlight the anticancer activity of Nigella sativa and it constitutes, focusing on different in vitro, in vivo and clinical studies and projects, in order to underline their antiproliferative, proapoptotic, cytotoxic and antimetastatic effects. Particular attention has been also given to the synergistic effect of Nigella sativa and it constitutes with chemotherapeutic drugs, and to the synthesized analogs of thymoquinone that seem to enhance the chemo-sensitizing potential. This review could be a useful step towards new research on N. sativa and cancer, to include this plant in the dietary treatments in support to conventional therapies, for the best achievement of therapeutic goals.

Immunoinflammatory effects of dietary bioactive compounds.

Inflammation is a key mechanism of the immune system that can be elicited by several factors, among them several chemical, physical and biological agents. Once stimulated, the inflammatory response activates a series of signaling pathways and a number of immune cells which promote, in a very coordinated manner, the neutralization of the infectious agent. However, if uncontrolled, the inflammatory status may become chronic leading, potentially, to tissue damage and disease onset. Several risk factors are associated with the development of chronic inflammation and, among these factors, diet plays an essential role. In this chapter the effects of some dietary bioactive compounds, including micronutrients, omega-3 fatty acids, nucleotides and polyphenols, on the immunoinflammatory responses in different cellular, animal and human studies have been summarized.

The roles of strawberry and honey phytochemicals on human health: A possible clue on the molecular mechanisms involved in the prevention of oxidative stress and inflammation.

BACKGROUND: Oxidative stress and inflammation contribute to the etiopathogenesis of several human chronic diseases, such as cancer, diabetes, cardiovascular diseases and metabolic syndrome. Besides classic stimuli, such as reactive oxidant species, endotoxins (i.e., bacteria lipopolysaccharide), cytokines or carcinogens, oxidative stress and inflammation can be triggered by a poor diet and an excess of body fat and energy intake. Strawberry and honey are common rich sources of nutrients and bioactive compounds, widely studied for their roles exerted in health maintenance and disease prevention. PURPOSE: This review aims to summarize and update the effects of strawberry and honey against oxidative stress and inflammation, with emphasis on metabolism and on the main molecular mechanisms involved in these effects. METHODS: A wide range of literature, published in the last 10 years, elucidating the effects of strawberry and honey in preventing oxidative stress and inflammation both in vitro (whole matrix and digested fractions) and in vivo was collected from online electronic databases (PubMed, Scopus and Web of Science) and reviewed. RESULTS: Strawberry and honey polyphenols may potentially prevent the chronic diseases related to oxidative stress and inflammation. Several in vitro and in vivo studies reported the effects of these foods in suppressing the oxidative stress, by decreasing ROS production and oxidative biomarkers, restoring the antioxidant enzyme activities, ameliorating the mitochondrial antioxidant status and functionality, among others, and the inflammatory process, by modulating the mediators of acute and chronic inflammation essential for the onset of several human diseases. These beneficial properties are mediated in part through their ability to target multiple signaling pathways, such as p38 MAPK, AMPK, PI3K/Akt, NF-κB and Nrf2. CONCLUSIONS: Available scientific literature show that strawberry and honey may be effective in preventing oxidative stress and inflammation. The deep evaluation of the factors that affect their metabolism as well as the assessment of the main molecular mechanisms involved are of extreme importance for the possible therapeutic and preventive benefit against the most common human diseases. However, published literature is still scarce so that deeper studies should be performed in order to evaluate the bioavailability of these food matrices and their effects after digestion.

Potential Health Benefit of Garlic Based on Human Intervention Studies: A Brief Overview.

Garlic is a polyphenolic and organosulfur enriched nutraceutical spice consumed since ancient times. Garlic and its secondary metabolites have shown excellent health-promoting and disease-preventing effects on many human common diseases, such as cancer, cardiovascular and metabolic disorders, blood pressure, and diabetes, through its antioxidant, anti-inflammatory, and lipid-lowering properties, as demonstrated in several in vitro, in vivo, and clinical studies. The present review aims to provide a comprehensive overview on the consumption of garlic, garlic preparation, garlic extract, and garlic extract-derived bioactive constituents on oxidative stress, inflammation, cancer, cardiovascular and metabolic disorders, skin, bone, and other common diseases. Among the 83 human interventional trials considered, the consumption of garlic has been reported to modulate multiple biomarkers of different diseases; in addition, its combination with drugs or other food matrices has been shown to be safe and to prolong their therapeutic effects. The rapid metabolism and poor bioavailability that have limited the therapeutic use of garlic in the last years are also discussed.

Phenolic compounds from Mediterranean foods as nutraceutical tools for the prevention of cancer: The effect of honey polyphenols on colorectal cancer stem-like cells from spheroids.

Chemoresistance and development of relapses are ascribable to a rare cell population of tumour mass: cancer stem cells (CSCs). Targeting CSCs could increase patients' survival rate and it is important to identify molecules that can act on the main pathways of these cells. Natural bioactive compounds, of which Manuka honey (MH) is rich, could be a good opportunity to target them. This work aims to evaluate the effect of MH on CSCs-like from human colorectal carcinoma (HCT-116 cell line) enriched through the in vitro sphere-forming assay. The results showed that MH reduced the volume of the entire culture of spheroids, affecting also their morphological parameters and induced apoptosis and ROS intracellular accumulation in CSCs-like. In addition, MH decreased the mRNA expression of one of ABC transporters (ABCG2) and affected self-renewal ability through the downregulation of the mRNA expression of one of the receptor membranes of Wnt/ß-catenin pathway (Frizzled 7).

Strawberry (Fragaria × ananassa cv. Romina) methanolic extract promotes browning in 3T3-L1 cells.

In recent years, the conversion of white adipocytes to brown-like adipocytes by pharmacological and dietary compounds has gained attention as an effective strategy to fight obesity. Strawberry bioactive compounds present several biological activities including antioxidant, anti-inflammatory, anti-cancer, anti-atherosclerotic and antiadipogenic properties. However, to the best of our knowledge, the possible role of strawberry bioactive compounds in white adipose tissue (WAT) browning has never been explored. Our results demonstrated that a strawberry methanolic extract (SE) significantly reduced 3T3-L1 pre-adipocytes differentiation, and down-regulated the mRNA expression of the adipogenic transcription factors CCAAT/enhancer-binding protein (C/REB- α) and peroxisome proliferation-activated receptor (PPAR-γ). It also down-regulated the mRNA expression of resistin and angiotensinogen, two genes considered as markers of white adipocytes, while increased the mRNA expression of pyruvate dehydrogenase lipoamide kinase isozyme 4 (PDK4) and uncoupling protein 1 (UCP1) which, conversely, are brown adipocyte-specific markers. Likewise, SE stimulated AMP-activated protein kinase (AMPKα), sirtuin 1 (Sirt1) and the peroxisome proliferator activated receptor gamma coactivator 1-alpha (PGC-1α), suggesting a possible increase in mitochondrial biogenesis. It also stimulated oxygen consumption rate and uncoupled respiration. Taken together, all these results suggest that SE induces brown fat-like phenotype in 3T3-L1 cells and may have potential therapeutic implications for treatment and/or prevention of obesity.

Resveratrol inhibits the proliferation of melanoma cells by modulating cell cycle.

The aim of this study was to evaluate the inhibitory effects of resveratrol (RSV) in A375 and A431 melanoma cells, by assessing cell viability (CCK-8 assay), apoptosis through flow cytometer and cell morphology, cell cycle assay by flow cytometer and western blot (Cyclin D1, Rac1 and PCDH9). Our results demonstrated that RSV strongly inhibited A375 cells proliferation, by decreasing cell viability, promoting apoptosis and arresting cell cycle. Besides, to clarify the main factor - duration or concentration of RSV, the double variance analysis of independent factors was operated after Bartlett's test for homogeneity by R project. According to the outcomes obtained from statistical analyses, Cyclin D1 and PCDH9 were strongly affected by RSV duration while Rac1 was not influenced. In conclusion, RSV can inhibit A375 proliferation and trigger apoptosis by influencing cell cycle proteins; for these effects, treatment duration of RSV played more important role than concentration.