Research background. Kombucha is consumed worldwide for its beneficial health effects. Kombucha teas fermented with various herbal infusions have become very important nowadays. Although black tea is used for kombucha fermentation, kombucha teas fermented with different herbal infusions have gained great importance. In this study, three different traditional medicinal plants, namely hops (Humulus lupulus L.), madimak (Polygonum cognatum), and hawthorn (Crataegus monogyna) were used for the fermentation of kombucha beverages, and the bioactivity of these beverages was investigated extensively. Experimental approach. The microbiological profile, bacterial cellulose formation, antibacterial and antiproliferative activity, antioxidant activities, sensory properties, total phenolic content, and flavonoid content of kombucha beverages were investigated. Liquid chromatography-coupled mass spectrometry analysis (LC-MS / MS) was used to identify and quantify specific polyphenolic compounds in the samples. Results and conclusions. According to the results, the hawthorn-flavoured kombucha, which has lower free radical scavenging activity than the other samples, came into prominence in terms of sensory properties. All kombucha beverages examined showed a strong cytotoxic effect on Mahlavu and HCT116 cell lines, but only the madimak-flavoured kombucha sample, which had a higher total phenolic/flavonoid content, demonstrated antibacterial activity against all microorganisms used in the study. Novelty and scientific contribution. Considering the results of this study, madimak could be an effective herb for the development of new kombucha beverages, although its sensory properties still need to be improved. This study contributes to science in terms of producing new fermented beverages with improved beneficial health effects.
Epidermal growth factor receptor (EGFR) has critical roles in epithelial cell physiology. Over-expression and over-activation of EGFR have been implicated in diverse cancers, including triple-negative breast cancers (TNBCs), prompting anti-EGFR therapies. Therefore, developing potent therapies and addressing the inevitable drug resistance mechanisms necessitates deciphering of EGFR related networks. Here, we describe Sorting Nexin 3 (SNX3), a member of the recycling retromer complex, as a critical player in the epidermal growth factor (EGF) stimulated EGFR network in TNBCs. We show that SNX3 is an immediate and sustained target of EGF stimulation initially at the protein level and later at the transcriptional level, causing increased SNX3 abundance. Using a proximity labeling approach, we observed increased interaction of SNX3 and EGFR upon EGF stimulation. We also detected colocalization of SNX3 with early endosomes and endocytosed EGF. Moreover, we show that EGFR protein levels are sensitive to SNX3 loss. Transient RNAi models of SNX3 downregulation have a temporary reduction in EGFR levels. In contrast, long-term silencing forces cells to recover and overexpress EGFR mRNA and protein, resulting in increased proliferation, colony formation, migration, invasion in TNBC cells, and increased tumor growth and metastasis in syngeneic models. Consistent with these results, low SNX3 and high EGFR mRNA levels correlate with poor relapse-free survival in breast cancer patients. Overall, our results suggest that SNX3 is a critical player in the EGFR network in TNBCs with implications for other cancers dependent on EGFR activity.
Sorting Nexin 3 (SNX3) gene maps to chromosome 6, minus strand and has 4 exons and 3 introns. There are 3 alternatively spliced isoforms (transcripts). SNX3 is a member of the sorting nexin family. Members of this family generally have BAR domains and phosphoinositide binding regions called the phox (PX) domain, and are involved in intracellular trafficking. Unlike other sorting nexins, SNX3 does not contain a BAR domain. SNX3 protein interacts with phosphatidylinositol-3phosphates, and is involved in protein trafficking through its role in the retromer complex.
ERBB family member epidermal receptor tyrosine kinase (EGFR) is composed of 28 exons and 27 introns. EGFR codes for 11 transcripts and 8 of them are protein coding. EGFR is a transmembrane glycoprotein that can be activated by several different ligands such as epidermal growth factor (EGF), transforming growth factor-alpha (TGFA), heparin-binding EGF-like growth factor (HBEGF), betacellulin (BTC), amphiregulin (AREG), epiregulin (EREG), and epigen (EPGN) (Singh, 2016). Ligand binding induces the dimerization of EGFR and autophosphorylation followed by a cascade of downstream phosphorylation events (Capuani et al., 2015). EGFR activation plays a key role in cell survival, proliferation, migration and differentiation (Purba, 2017).
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