MALAT-1/p53/miR-155/miR-146a ceRNA circuit tuned by methoxylated quercitin glycoside alters immunogenic and oncogenic profiles of breast cancer.

Triple-Negative Breast Cancer (TNBC) is one of the most aggressive and hot BC subtypes. Our research group has recently shed the light on the utility of natural compounds as effective immunotherapeutic agents. The aim of this study is to investigate the role of a methoxylated quercetin glycoside (MQG) isolated from Cleome droserifolia in harnessing TNBC progression and tuning the tumor microenvironment and natural killer cells cytotoxicity. Results showed that MQG showed the highest potency (IC50 = 12 µM) in repressing cellular proliferation, colony-forming ability, migration, and invasion capacities. Mechanistically, MQG was found to modulate a circuit of competing endogenous RNAs where it was found to reduce the oncogenic MALAT-1 lncRNA and induce TP53 and its downstream miRNAs; miR-155 and miR-146a. Accordingly, this leads to alteration in several downstream signaling pathways such as nitric oxide synthesizing machinery, natural killer cells' cytotoxicity through inducing the expression of its activating ligands such as MICA/B, ULBP2, CD155, and ICAM-1 and trimming of the immune-suppressive cytokines such as TNF-α and IL-10. In conclusion, this study shows that MQG act as a compelling anti-cancer agent repressing TNBC hallmarks, activating immune cell recognition, and alleviating the immune-suppressive tumor microenvironment experienced by TNBC patients.

Niazimicin: A thiocarbamate glycoside from Moringa oleifera Lam. seeds with a novel neuroprotective activity.

Moringa oleifera (MO) known as the miracle tree is a famous nutritional source in many countries. In this study, the neuroprotective activity of MO seeds was investigated. Fractions of the 70% ethanol seed extract of MO were injected at a dose of 250 mg kg-1  day-1 to albino rats for 15 days, after-which induction of dementia was done using 100 mg/kg AlCl3 over 30 days. Results revealed that all fractions ameliorated the effects of AlCl3 where methylene chloride and ethyl acetate fractions, containing the major bioactive compound niazimicin (NZ), showed the best activities. Biological investigations proved NZ to be a highly potent neuroprotective drug lead as a first report, by causing a decrease in the levels of malondialdehyde, cholinesterase, nitric oxide (NO) and amyloid ß by 47%, 34%, 53% and 59%, respectively, and increasing glutathione levels by 54%. Molecular docking studies suggested NZ neuroprotective effects to be mediated by inhibition of caspase-3 and inducible nitric oxide synthase enzymes. PRACTICAL APPLICATIONS: The current findings present the neuroprotective effect of Moringa oleifera seeds consumed as a food supplement and in daily diet. In addition, niazimicin is a promising lead for the development of novel agents against Alzheimer's disease as seen by the reported results.

Targeting hydrogen sulphide signaling in breast cancer.

INTRODUCTION: Hydrogen sulphide (H2S) has been established as a key member of the gasotransmitters family that recently showed a pivotal role in various pathological conditions including cancer. OBJECTIVES: This study investigated the role of H2S in breast cancer (BC) pathogenesis, on BC immune recognition capacity and the consequence of targeting H2S using non-coding RNAs. METHODS: Eighty BC patients have been recruited for the study. BC cell lines were cultured and transfected using validated oligonucleotide delivery system. Gene and protein expression analysis was performed using qRT-PCR, western blot and flow-cytometry. In-vitro analysis for BC hallmarks was performed using MTT, BrdU, Modified Boyden chamber, migration and colony forming assays. H2S and nitric oxide (NO) levels were measured spectrophotometrically. Primary natural killer cells (NK cells) and T cell isolation and chimeric antigen receptor transduction (CAR T cells) were performed using appropriate kits. NK and T cells cytotoxicity was measured. Finally, computational target prediction analysis and binding confirmation analyses were performed using different software and dual luciferase assay kit, respectively. RESULTS: The H2S synthesizing enzymes, cystathionine ß-synthase (CBS) and cystathionine γ-lyase (CSE), exhibited elevated levels in the clinical samples that correlated with tumor proliferation index. Knock-down of CBS and CSE in the HER2+ BC and triple negative BC (TNBC) cells resulted in significant attenuation of BC malignancy. In addition to increased susceptibility of HER2+ BC and TNBC to the cytotoxic activity of HER2 targeting CAR T cells and NK cells, respectively. Transcriptomic and phosphoprotein analysis revealed that H2S signaling is mediated through Akt in MCF7, STAT3 in MDA-MB-231 and miR-155/ NOS2/NO signaling in both cell lines. Lastly, miR-4317 was found to function as an upstream regulator of CBS and CSE synergistically abrogates the malignancy of BC cells. CONCLUSION: These findings demonstrate the potential role of H2S signaling in BC pathogenesis and the potential of its targeting for disease mitigation.

An acetylated derivative of vitexin halts MDA-MB-231 cellular progression and improves its immunogenic profile through tuning miR- 20a-MICA/B axis.

The activating immune ligands, MICA/B, act as a "kill me" signal through the NKG2D receptor expressed on natural killer (NK) cells. Recently, the oncogenic miR-20a was found to mediate immune escape through repressing MICA/B levels in breast cancer (BC) cells. However, targeting miR-20a-MICA/B using natural compounds has rarely been investigated. Our group has successfully isolated 3'-O-acetylvitexin that showed cytotoxic effects against colon cancer cells but has never been evaluated in BC. Our aim is to investigate the effects of 3'-O-acetylvitexin on BC cell lines and to further elucidate its molecular mechanism of action.The results showed that 3'-O-acetylvitex depicted a more pronounced dose-dependent repression of TNBC cellular viability, colonogenicity and migration capacity than Vitexin. 3'-O-acetylvitexin treatment resulted in a marked dose-dependent repression of miR-20a with a concomitant dose-dependent increase in MICA/B expression. In conclusion, 3'-O-acetylvitexin might act as a promising therapeutic agent for TNBC patients.

Upregulation of MC4R and PPAR-α expression mediates the anti-obesity activity of Moringa oleifera Lam. in high-fat diet-induced obesity in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: various extracts of Moringa oleifera Lam. leaves, were reported to possess antiobesity effect in experimental animals models, yet its active doses and mechanism of action are still unclear. MATERIALS AND METHODS: The metabolic profiling of 70% ethanol extract of M. oleifera (MO) leaves was performed using HPLC-MS/MS analysis. The antiobesity activity of MO was tested in high fat diet induced obesity in rats at 200 and 400 mg/kg body weight orally for 1 month. Total cholesterol (TC), high density lipoproteins (HDL-C), low density lipoprotein-cholesterol (LDL-C), triglycerides (TGs), insulin resistance, insulin sensitivity, and adipose tissue index were monitored. In addition, fatty acid synthase (FAS) and HMG-CoA reductase mRNA from liver tissue, Peroxisome Proliferator-Activated Receptor alpha (PPARα) and Melanocortin-4 receptor (MC4R) RNA from adipose tissue were quantified using qRT-PCR. MO hard gelatin capsules (400 mg/capsule) were formulated and standardized using HPLC-RP analysis and tested on fifteen female participants, aged 45-55 with a BMI of 29-34 kg/m2. RESULTS: Thirteen metabolites were tentatively identified using HPLC-MS/MS analysis including flavonols, flavones and a phenolic acid. MO 400 showed a prominent effect on reducing the rats' final weights, % weight increase and adiposity index (P < 0.05). Glucose, insulin and HOMA-IR were significantly reduced and R-QUICKI was significantly increased by MO 400 (P < 0.001). Mean tissue level of leptin and vaspin were significantly reduced, adiponectin, omentin and GLUT-4 expression were increased significantly by MO 400 (P < 0.01). MO 400 significantly suppressed FAS and HMG-CoA reductase and increased mRNA expression of MC4R and PPAR-α (P < 0.01). Eight weeks administration of MO hard gelatin capsules to obese patients showed significant reduction of the average BMI, TC and LDL compared to the baseline values (p < 0.05). CONCLUSION: Our results presented a scientific evidence for the traditional use of M. oleifera leaves as antiobesity herbal medicine.

Artichoke edible parts are hepatoprotective as commercial leaf preparation

ABSTRACT Chemical profile analyses of artichoke (Cynara scolymus L., Asteraceae) edible parts (fleshy receptacle, inner bracts) as well as roots are compared with the commercially usable leaf extract using HPLC-DAD-ESI-MS via chlorogenicacid as a marker. Overall polyphenolic constituents demonstrated by means of LC/MS profiling. The nutritional values and inulin contents of different assessed parts were investigated. The present study was designed to determine the effect of artichoke: leaves, bracts, receptacles and roots alcoholic extracts against CCl4-induced acute hepatotoxicity and hyperlipidemia in rats by means of histopathological and biochemical parameters. Serum liver enzymes levels of aspartate amino transferase, alanine amino transferase, alkaline phosphatase and lipid peroxidase content (malondialdehyde MDA) were estimated. Blood glutathione, total cholesterol, triacylglycerides and high density lipid level were estimated in plasma. The ethanol extract of roots, leaves, bracts and receptacles were standardized to (0.82 ± 0.02, 1.6 ± 0.06, 2.02 ± 0.16 and 2.4 ± 0.27 mg chlorogenic acid/100 mg extract), respectively. The receptacle showed the highest content of polyphenols and exhibits the highest antioxidant activity. HPLC analysis of inulin in the receptacles of globe artichoke revealed high content of inulin (41.47 mg/g) dry extract. All artichoke parts contain comparable vitamins and minerals. Artichokes receptacles extract when taken in dose of (500 mg/kg/day) reduce the lesion caused by CCl4 alone more than groups receiving silymarin. Bracts and leaves extract exert nearly the same effect.

Cytotoxic activity of phenolic constituents from Echinochloa crus-galli against four human cancer cell lines

Abstract Echinochloa crus-galli (L.) P. Beauv., Poaceae, grains are used as a feed for birds and millet for humans. The sulforhodamine B assay was used to assess its cytotoxicity against four human cancer cell lines. The ethanolic extract (70%) proved to be most active against HCT-116 and HELA cell lines (IC50 = 11.2 ± 0.11 and 12.0 ± 0.11 µg/ml, respectively). On the other hand, the chloroform and ethyl acetate fractions exhibited their highest activities against HCT-116 cell lines. The chloroform and ethyl acetate fractions were subjected to several chromatographic separations to render pure phenolic compounds (1-8). Compounds 1-8 were identified as: 5,7-dihydroxy-3′,4′,5′-trimethoxy flavone, 5,7,4′-trihydroxy-3′,5′-dimethoxy flavone (tricin), quercetin, flavone, apigenin-8-C-sophoroside, 2-methoxy-4-hydroxy cinnamic acid, p-coumaric acid and quercetin-3-O-glucoside. All the isolated phenolic compounds exhibited various significant activities against the four human carcinoma where the methoxylated flavones (1 and 2) were the most active, in a way comparable to the anticancer drug Doxorubicin®. Thus, these methoxylated flavonoids may be considered as lead compounds for the treatment of cancer, which supports previous claims of E. crus-galli traditional use. This is the first report of the occurrence of these phenolic compounds in E. crus-galli.