Piceatannol induces caspase-dependent apoptosis by modulating intracellular reactive oxygen species/mitochondrial membrane potential and enhances autophagy in neuroblastoma cells.

The aim of this study was to evaluate the anticancer effects of piceatannol, a natural stilbenoid, on human neuroblastoma cells. In order to accomplish this goal, we performed various cellular assays, including the XTT cell proliferation assay for cell viability, colony formation assay for colony formation capacity, FITC Annexin V and cell death detection kit for apoptosis, matrigel invasion assay for invasion capacity, intracellular reactive oxygen species (ROS) red dye for intracellular ROS levels, TMRM staining method for mitochondrial membrane potential (MMP), and the CYTO-ID autophagy detection kit for autophagy. Furthermore, we analyzed the expression levels of genes associated with apoptosis and autophagy using RT-qPCR. Based on our findings, piceatannol exhibited cytotoxic effects on neuroblastoma cells. Besides, treatment with piceatannol at both 50 and 100 µM concentrations for 72 h decreased colony formation, induced apoptosis and autophagy, inhibited cell invasion, decreased MMP, and increased ROS levels in SH-SY5Y cells. In addition, we observed significant upregulation in the expression levels of CASP8, BECLIN, ATG5, ATG7, and MAPILC3A genes between the two doses. These results suggest that piceatannol enhances autophagic activity and induces caspase-dependent apoptosis, indicating its potential as a therapeutic agent against neuroblastoma cells.

Lysates from the probiotic bacterium Streptococcus thermophilus enhances the survival of T cells and triggers programmed cell death in neuroblastoma cells.

Neuroblastoma is the most common brain solid tumor in infancy. Despite the availability of numerous approaches like immunotherapy, surgery, chemotherapy, and radiotherapy, neuroblastoma frequently develops resistance and recurs. Immunotherapy is one of the most promising approaches and PD-L1 antibody blocking is the phenomena used to inhibit PD-1 receptors to increase and improve cytotoxic T cells toward cancer. Numerous studies underlined the critical role of probiotics on immune system development and modulation in addition to possible role in inducing apoptosis in cancer cells. In this study, a Streptococcus thermophilus strain, isolated from a local yogurt, was used as it is considered a potential probiotic due to its tolerance lower pH, bile acid, antibiotic suitability, and blood hemolysis. Our results showed that S. thermophilus lysates played as an immune checkpoint modulator at 25 µg/ml dose boosting PD-L1 transcripts and protein levels in SH-SY5Y neuroblastoma cell line. Interestingly, co-culture between SH-SY5Y and Jurkat T cells in the presence of blocking PD-L1 antibodies increased Jurkat T-cell viability compering to control without lysate. On the other hand, annexin-V/7-AAD, qPCR and western blot results showed that S. thermophilus lysates at 200 and 400 µg/ml decreased SH-SY5Y cell viability and increased apoptotic marker genes transcription and caspase-3 and caspase-9 protein expression.

Autophagic mechanisms in longevity intervention: role of natural active compounds.

The term 'autophagy' literally translates to 'self-eating' and alterations to autophagy have been identified as one of the several molecular changes that occur with aging in a variety of species. Autophagy and aging, have a complicated and multifaceted relationship that has recently come to light thanks to breakthroughs in our understanding of the various substrates of autophagy on tissue homoeostasis. Several studies have been conducted to reveal the relationship between autophagy and age-related diseases. The present review looks at a few new aspects of autophagy and speculates on how they might be connected to both aging and the onset and progression of disease. Additionally, we go over the most recent preclinical data supporting the use of autophagy modulators as age-related illnesses including cancer, cardiovascular and neurodegenerative diseases, and metabolic dysfunction. It is crucial to discover important targets in the autophagy pathway in order to create innovative therapies that effectively target autophagy. Natural products have pharmacological properties that can be therapeutically advantageous for the treatment of several diseases and they also serve as valuable sources of inspiration for the development of possible new small-molecule drugs. Indeed, recent scientific studies have shown that several natural products including alkaloids, terpenoids, steroids, and phenolics, have the ability to alter a number of important autophagic signalling pathways and exert therapeutic effects, thus, a wide range of potential targets in various stages of autophagy have been discovered. In this review, we summarised the naturally occurring active compounds that may control the autophagic signalling pathways.

Piceatannol induces apoptotic cell death through activation of caspase-dependent pathway and upregulation of ROS-mediated mitochondrial dysfunction in pancreatic cancer cells.

BACKGROUND: Piceatannol is a naturally occurring plant-derived phenolic compound (stilbenoid), an analogue of resveratrol. It has been shown that, piceatannol has biological activity properties such as antiproliferative, antioxidative, anti-inflammatory and proapoptotic, in various human cancer studies in vitro and in vivo. OBJECTIVES AND METHODS: In this study, it was aimed to investigate whether piceatannol induces apoptosis through anticancer activity methods (cell viability, colony formation, annexin-V/7-AAD, ROS (Reactive oxygen species), MMP (Mitochondrial membrane potential), wound healing, invasion assay, RT-qPCR (Real-Time Quantitative Polymerase Chain Reaction), western blotting in PANC-1 and MIA PaCa-2 pancreatic cancer (PC) cell lines. RESULTS: According to our results, piceatannol decreased cell viability in a dose and time-dependent manner [the half-maximal inhibitory concentration (IC50): 60 µM in PANC-1 and IC50: 90 µM in MIA PaCa-2 cell line at 48 h (h)] and caused significant changes in the expression of apoptosis-related genes and protein. Piceatannol induced apoptosis in PANC-1 and MIA PaCa-2 cells, accompanied by increased ROS production, decreased MMP, and increased Caspase-3-9 activity. Piceatannol also inhibited colony-forming abilities, invasion, and migration of PC cells. CONCLUSION: Our results show that piceatannol has an anti-cancerogenic effect on PANC-1 and MIA PaCa-2 cells, and exerts this effect by suppressing proliferation and inducing apoptosis. Therefore, piceatannol could be considered to be a potential chemotherapeutic agent candidate for the treatment and prevention of PC.

Tomentosin induces apoptosis in pancreatic cancer cells through increasing reactive oxygen species and decreasing mitochondrial membrane potential.

The aim of this study was to determine possible anticancer effect of tomentosin, a natural sesquiterpene lactone, on pancreatic cancer cells. The cytotoxic effect of tomentosin was determined by XTT analysis. Colony formation and apoptosis analyzes were performed, Reactive oxygen species (ROS) level and change in mitochondrial membrane potential (MMP) were evaluated in control and tomentosin-treated cells. The effect of tomentosin on expression levels of apoptosis-related genes was determined by qRT-PCR and Caspase-3 and Caspase-9 proteins were analyzed by western blot. And, the effect of tomentosin on migration and invasion of cells were evaluated. The IC50 dose of tomentosin was found to be 31.11 µM in PANC-1 cells and 33.93 µM in MIA PaCa-2 cells for 48 h. And, treatment of tomentosin at IC50 dose suppressed the colony forming capacity of cells. While tomentosin increased apoptosis rate and ROS production, an decrease was observed in MMP. Tomentosin affected expression level of apoptosis-related genes and increased Caspase-3 and Caspase-9 protein levels. After tomentosin treatment, cell migration and invasion were suppressed. As a result, this study reveals that tomentosin has anticancer effects on pancreatic cancer cells, and therefore it predicts that tomentosin can be evaluated as an effective agent against pancreatic cancer.

Anticancer Effect and Phytochemical Profile of the Extract from Achillea ketenoglui against Human Colorectal Cancer Cell Lines.

BACKGROUND: In the treatment of Colorectal Cancer (CRC), the search for new antineoplastic drugs with fewer side effects and more effectiveness continues. A significant part of these pursuits and efforts focus on medicinal herbs and plant components derived from these plants. A. ketenoglui is one of these medicinal plants, and its anticancer potential has never been studied before. METHODS: The phenolic and flavonoid content, and antioxidant activity of A. ketenoglui extracts were determined. The phytochemical profiling and quantification analysis of major components were performed by HPLC-ESI-Q-TOF-MS. Cytotoxicity, proliferation, apoptosis and cell cycle were evaluated to reveal the anticancer activity of the extract on CRC cells (HCT 116 and HT-29). The determined anticancer activity was confirmed by mRNA (RT-qPCR) and protein (Western blotting) analyzes. RESULTS: A. ketenoglui methanol extract was found to have high phenolic (281.89±0.23) and flavonoid (33.80±0.15) content and antioxidant activity (IC50 40.03±0.38). According to the XTT assay, the extract has strong cytotoxic activity (IC50 350 µM in HCT 116 and IC50 263 µM in HT-29 cell line). The compounds most commonly found in the plant are, in descending order, chlorogenic acid, apigenin, genistin, baicalin, eupatorin, casticin, and luteolin. In flowcytometric analysis, the extract was found to induce greater apoptosis and cell cycle arrest in both cell lines than in both control and positive control (casticin). According to the results of the mRNA expression analysis, the extract treatment upregulated the expression of the critical genes of the cell cycle and apoptosis, such as p53, p21, caspase-3, and caspase-9. In protein expression analysis, an increase in caspase-3 and p53 expression was observed in both cell lines treated with the extract. In addition, caspase-9 expression was increased in HT-29 cells. CONCLUSION: The findings show that A. ketenoglui has an anticancer potential by inducing apoptosis and arresting the cancer cell cycle and may be promising for CRC therapy. This potential of the plant is realized through the synergistic effects of its newly identified components.

The Phytochemical Profile and Biological Activity of Liquidambar orientalis Mill. var. orientalis via NF-κB and Apoptotic Pathways in Human Colorectal Cancer.

Liquidambar orientalis Mill. var. orientalis (Hammamelidaceae) commonly known as oriental sweet gum is a medicinal plant endemic to Turkey, and used for treatment of wounds such as ulcers, gastritis and skin diseases. However, there are only a limited number of studies on the anticancer properties of this plant. The aim of this study is to investigate the cellular and molecular mechanisms of L. orientalis in colorectal cancer cell lines and to reveal the active therapeutic components. Antioxidant, anti-proliferative, anti-invasive and apoptotic effects were investigated for determining biological activities. The phytochemical profile and quantitation assays were carried out by using HPLC-ESI-Q-TOF-MS. The results demonstrated that leaf methanol extract (LM) of L. orientalis has the highest cytotoxic activity in HCT-116 (IC50 27.80 µg/mL) and HT-29 (IC50 43.13 µg/mL) cell lines as compared to the other extracts tested. Regarding chemical composition, quercetin 3-glucoside (61.005 ± 1.527 mg/g extract), chlorogenic acid (31.627 ± 0.970 mg/g extract), pyrogallol (9.950 ± 0.001 mg/g extract), epigallocatechin gallate (9.671 ± 0.004 mg/g extract), apigenin 7-O-glucoside (2.687 ± 0.027 mg/g extract), gallic acid (2.137 ± 0.012 mg/g extract), genistin (1.270 ± 0.027 mg/g extract), luteolin (0.055 ± 0.0005 mg/g extract) and kaempferol (0.506 ± 0.007 mg/g extract) were identified as the major phytochemical compounds. Our results showed that LM extract exhibited In Vitro cytotoxic and apoptotic properties.

Evaluation of Apoptosis Pathway of Geraniol on Ishikawa Cells.

Endometrial cancer is the most common type of cancer in the female reproductive system. Geraniol is acyclic monoterpene alcohol derived from essential oils of aromatic plants. This study aimed to investigate the apoptosis pathway of geraniol on Ishikawa cells. The cytotoxic effects of Geraniol on Ishikawa cells were determined by an MTT test. Ishikawa cells were seeded on cover slips, the IC50 dose was applied, and the cells were incubated with antibodies against Bax, Bcl-2, and TUNEL Assay. mRNA expression analysis of apoptosis-related genes was determined by RT-qPCR with an IC50 dose of Geraniol. The IC50 dose of Geraniol decreased Bcl-2 staining significantly, but it significantly increased Bax staining and TUNEL positive cells. A significant increase in the Bax, caspase3, caspase-8, cytochrome C and Fas genes and a significant decrease in the Bcl-2 gene was observed when the IC50 dose group was compared to the cells in the control group based on their mRNA expression levels.Analysis of expression of genes whose products are involved in apoptosis suggests the involvement of the mitochondrial pathway.

Bioactive Compounds of Rheum ribes L. and its Anticancerogenic Effect via Induction of Apoptosis and miR-200 Family Expression in Human Colorectal Cancer Cells.

Rheum ribes L. is a widespread plant species in the eastern part of our country and is consumed as a vegateble by the folk people. In our study, we investigated whether R. ribes has anti-cancerogenic effect on colorectal cancer cell lines. Six different extracts were obtained from R. ribes. Total phenolic, flavonoid contents, antioxidant activities, and cytotoxic effects of all extracts on colorectal cancer cells were determined. Differences in expression levels of the miR-200 family and target genes were assessed by real-time qPCR in cells. Antiproliferative effect was determined by TUNEL, Annexin V, cell cycle, invasion analysis; and levels of BCL-2, ZEB1, GATA4 and FAS/CD95 proteins were determined by ELISA. According to the results of miRNA expression analysis, the only dose group with a significant increase (p < 0,05) in the expression level of all of miRNAs in both cell lines was RM (Root methanol) extract alone. Our results showed that, RM caused a significant increase in the expressions of miR-200a/b/c and miR-141, and it suppressed BCL-2, ZEB1, GATA4 expressions, with this way. Thus, it has been suggested that the treatment of RM alone, or combined treatment of RM with 5-FU result in a better response than cancer cells treated with 5-FU alone.

Homoarginine, ß-ODAP, and asparagine contents of grass pea landraces cultivated in Turkey.

The seeds of grass pea (Lathyrus sativus L.), a drought tolerant crop, were analysed for quantitative determination of the free amino acids ß-N-oxalyl-l-α,ß-diaminopropionic acid (ß-ODAP), homoarginine and asparagine by a simple and fast capillary electrophoretic method. In boric acid (80mM, pH 8.0) running buffer system, not only were α and ß-ODAP successfully separated, but also an efficient sample stacking was achieved during hydrodynamic sample introduction. The validated method was used for quantification of ß-ODAP, homoarginine and asparagine in seed extracts of 52 Lathyrus local landraces from various regions of Turkey and one released cultivar. The concentration ranges of amino acids were found as 0.21-1.27% (w/w) for homoarginine, 0.10-0.87% (w/w) for ß-ODAP and 0.006-0.47% (w/w) for asparagine. A positive correlation between homoarginine and ß-ODAP quantities in seeds of 53 Lathyrus local landraces was shown to exist (r(2)=0.649).

Biodiversity in morphological properties and nutritional values of forage grass species.

This study was conducted under humid and warm climatic conditions in 19 Ondokuz Mayis University, Kurupelit campus area (1300 ha) during March and July in 2004. The study aimed to evaluate the forage grass species, which have been protected for over 30 years, and to observe the biological diversity by determining their morphological characters and forage qualities. Some morphological characters and chemical properties of 20 grass species were examined. Statistically significant differences were determined regarding all morphological characters and chemical properties within species and individual species. In examined grass species, number of tiller per plant and main stem length ranged from 5.5 to 40.5 and 39.38 to 96.18 cm, respectively. Root crown diameter, dry matter of root, dry forage weight changed between 6.24 and 21.60 mm, 0.27 and 20.33 g plant(-1), 0.80 and 46.76 g plant(-1), respectively. Protein content of the samples varied from 4.37 to 9.42%. Trace elements content of the samples such as Ca (0.08-0.79%), Mg (0.06-0.17%) and K (0.76-3.03%) were also determined. Potassium contents of dry grass forages were excessively high, however their phosphor contents were generally insufficient, calcium and magnesium contents were fairly insufficient and K/Ca+Mg ratios were over the critic value (2.20) except for two species. In general, Fe, Zn and Mn in dry grass forage samples were found adequate.