Whole Extracts Research Articles

Dark Sweet Cherry (Prunus avium) Phenolics Enriched in Anthocyanins Induced Apoptosis in MDA-MB-453 Breast Cancer Cells through MAPK-Dependent Signaling and Reduced Invasion via Akt and PLCγ-1 Downregulation

Dark sweet cherries (DSCs) are rich source of phenolics known to exert anticancer and anti-invasive activities. This study elucidated the molecular mechanisms underlying the activity of DSC phenolics against MDA-MB-453 breast cancer cells In Vitro. Cells were treated with DSC phenolics in whole extract (WE), and fractions enriched in anthocyanins (ACN) and proanthocyanidins (PCN) at concentrations that inhibited cell growth by 50%. Results showed that DSC phenolics suppressed Akt and PLCγ-1 activation, and inhibited cell motility and invasion, but only ACN reached significance. The extrinsic and intrinsic apoptotic pathways were also activated by DSC phenolics via caspase-8 cleavage and increased Bax/Bcl-2 ratio, with ACN exhibiting significant activation and stronger PARP-1 cleavage. Furthermore, sustained activation of mitogen-activated protein kinases (MAPKs) ERK1/2 and p38 was observed wherein ERK1/2 (U0126) and p38 (SB203580) inhibitors confirmed crosstalk ERK1/2-Akt and MAPK intrinsic mitochondrial pathways. In conclusion, DSC phenolics inhibited MDA-MB-453 breast cancer cells by targeting cell signaling pathways that induce apoptosis and suppress cell invasion, with ACN showing enhanced chemopreventive activities.

Dark sweet cherry (Prunus avium) phenolics enriched in anthocyanins exhibit enhanced activity against the most aggressive breast cancer subtypes without toxicity to normal breast cells

Abstract This study assessed dark sweet cherry (DSC) phenolics as chemopreventive/chemotherapeutics against aggressive breast cancer subtypes. DSC phenolics in whole extract (WE) and fractions enriched in phenolic acids (F1), flavonols (F3) and proanthocyanidins (F4) inhibited breast cancer cell growth with potency MDA-MB-453 > MDA-MB-231 ∼ BT-474, without toxicity to MCF-10A normal breast cells, except F3 that inhibited BT-474 and MCF-10A cells with similar toxicity. DSC anthocyanins (F2) inhibited breast cancer cell lines with similar potencies without toxicity to MCF-10A cells. Mechanisms for WE, F2 and F4 breast anticancer activity may be mediated by reduction of oxidative stress, Akt/mTOR, p38, and survivin regulation, suggesting antiproliferative and apoptotic mechanisms. F2 significantly downregulated mRNA levels of invasive/metastatic biomarkers Sp1, Sp4, and VCAM-1, and Sp1 protein suggesting an enhanced chemopreventive/chemotherapeutic potential of DSC anthocyanins. Further studies are needed to elucidate underlying mechanisms of DSC phenolics as natural breast cancer chemopreventive compounds.

Dark Sweet Cherry (Prunus Avium L.) Phenolics Induce Apoptosis and Inhibit Invasion on Breast Cancer MDA-MB-453 Through MAPK-dependent and Independent Pathways (P05-007-19)

ObjectivesDark sweet cherry (DSC) contains a rich mixture of phenolics reported to exert anti-cancer and anti-metastasis activities. We aimed to investigate the underlying mechanisms by which DSC phenolics in whole extracts (WE) and fractions enriched in anthocyanins (ACN) or procyanidins (PCN) decrease proliferation and invasion on the HER2 + breast cancer MDA-MB-453 cells. MethodsMDA-MB-453 cells were treated with DSC phenolics in WE, ACN, and PCN obtained by solid phase extraction (SPE) at doses that inhibited cell growth by 50% for 24 h. Protein expression over 24 h was assessed by western blots and invasion and metastasis potential were assessed with cell invasion and cell motility assays. ResultsDSC phenolics in WE, ACN, and PCN targeted intrinsic and extrinsic apoptotic pathways as shown by caspase-8 cleavage and cytochrome c/bax upregulation that culminate in caspase-9, -3, and PARP cleavage. In addition, a sustained activation of mitogen activated protein kinases (MAPKs) pathway was induced through ERK1/2 and p-38 phosphorylation with order of potency ACN ≥ PCN > WE. The crosstalk of MAPK-apoptotic pathways were confirmed with ERK1/2 and p-38 inhibitors that partially abrogated cytochrome/bax upregulation. In addition, DSC phenolics downregulated phosphoinositide 3-kinase (PI3K) signaling pathway, which act as key regulator in cancer proliferation and metastasis, and was accompanied by cell motility inhibition. ERK1/2 and p-38 inhibitors did not reverse cell motility and invasion reduction exerted by DSC phenolics. ConclusionsDSC phenolics target pro-apoptotic intrinsic/extrinsic, and cell growth/invasion mechanisms involving PI3K and MAPK pathways. ACN showed the most consistent induction of apoptotic markers and contributed to MAPK sustained upregulation with potency ACN ≥ PCN > WE. This MAPK upregulation contributed to the intrinsic apoptotic pathway but not to the PI3K/proliferation/invasion pathways. Funding SourcesThis work was supported by the Northwest Cherry Growers.

Dark Sweet Cherry phenolics as Dietary Chemopreventive/therapeutic Compounds for Aggressive Breast Cancer Cell Growth with No Toxicity to Normal Breast Cells (FS13-03-19)

ObjectivesDark sweet cherries (DSC) (Prunus avium) contain phenolic compounds with chemopreventive and chemotherapeutic potential. This study aimed to evaluate the effect of DSC extracts in inhibiting the most aggressive breast cancer subtypes without toxicity to normal cells. MethodsDSC phenolic extracts (WE) or its fractions enriched in phenolic acids (F1), anthocyanins (F2), flavonols (F3) or procyanidins (F4) were assessed for their antiproliferative activity against the most aggressive breast cancer BT-474, MDA-MB-453, and MDA-MB-231 cell lines, and MCF-10A noncancer breast epithelial cells using the resazurin assay. Cell levels of oxidative stress were assessed with fluorescein derivative H2DFFDA. Expression of genes linked to cell growth, apoptosis, and metastasis was assessed. ResultsDSC phenolics in whole extract (WE) and fractions enriched in phenolic acids (F1), flavonols (F3) and procyanidins (F4) inhibited MDA-MB-453 with higher potency (lower IC50values) compared to the other cell lines tested (P < 0.05) followed by MDA-MB-231 ∼ BT-474, without toxicity to MCF-10A cells. F3 inhibited BT-474 and MCF-10A cells with similar potency (P > 0.05). F2 inhibited all assessed breast cancer cell lines with similar potencies without toxicity to MCF-10A cells. Mechanisms for WE, F2 and F4 inhibitory activity on MDA-MB-453 cells were mediated by reduction of ROS (down to 28%, 56%, and 58% of control, respectively); which mediate proliferation and other events required for tumor progression. This was accompanied by downregulation of Akt/mTOR, p38, and survivin mRNA, suggesting antiproliferative and apoptotic mechanisms. An enhanced chemopreventive/chemotherapeutic potential of DSC anthocyanins is suggested because mRNA levels of invasive/metastatic biomarkers Sp1, Sp4, and VCAM-1 were downregulated only by F2. ConclusionsThese results demonstrate the chemopreventive/chemotherapeutic potential of DSC phenolics and fractions enriched in anthocyanins and procyanidins as natural compounds to target cell pathways linked to proliferation, apoptosis and invasion in breast cancer. The underlying molecular mechanisms are currently under investigation. Funding SourcesThis work was supported by the Northwest Cherry Growers.

Evaluation of some neuropharmacological effects of Caladium bicolor aiton (araceae) leaf extracts in mice.

Caladium bicolor Aiton (Araceae) is used in ethnomedicine for the treatment of boils, wound ulcers and convulsion. This study investigated the effects of the leaf extracts on some neuropharmacological parameters. The leaves were collected, dried, powdered and then extracted by maceration in methanol to yield the whole extract (WE). Extraction was also done using n-hexane, ethyl acetate and methanol in a Soxhlet apparatus to obtain n-hexane (HE), ethyl acetate (EA) and methanol (ME) extracts. Preliminary phytochemical screening was done using the whole extract. Some neuropharmacological evaluations were carried out using standard methods. Phytochemical screening revealed the presence of carbohydrates, proteins, alkaloids and flavonoids. WE showed varying levels of protection against strychnine-induced convulsion. Each of HE, EA and ME increased latency (P < 0.01) to pentylenetetrazole-induced convulsion and offered varying levels of protection against maximal electroshock-induced seizure. Each of WE, HE and ME significantly increased the duration of stay on the open arm of the elevated plus maze. Both EA and ME at doses of 100 and 200mg/kg, and HE at a dose of 400mg/kg significantly reduced the duration of immobility in forced swim test. It is concluded that the leaf extracts possess anticonvulsant, anxiolytic and antidepressant properties.

Evaluation of pranthocyanidin enriched cranberry extract for carbohydrate hydrolysis enzyme inhibition (647.38)

Proanthocyanidins (PACs), found in cranberries (Vaccinium macrocarpon), are hypothesized to be natural, oral hypoglycemic agents. This study investigates the inhibition potential of cranberry powder in carbohydrate hydrolysis enzymes. The whole extract (WE) was achieved by dissolving one gram of cranberry powder in 100 mL acetone solution (acetone: water: hydrochloric acid, 70:29.9:0.01) at room temperature. The sample was differentiated into two fractions: low molecular weight phenolic enriched fraction (LMWF) and PAC enriched fraction (PACF) using LH‐20. PAC contents were evaluated by 4‐Dimethylaminocinnamaldehyde (DMAC)assay and yielded 1.35, 0.03 and 0.59 mg/mL for WE, LMWF and PACF respectively. The total phenolic contents of were determined to be 4.68, 0.60, and 2.51 mg/mL for WE, LMWF and PACF respectively. Rat α glucosidase assays showed PACF has 3 times lower inhibitory activity and LMWF has almost 2 times higher inhibitory activity, compared to WE. Sucrase inhibition assays showed PACF has lower inhibitory activity and that LMWF has almost 2 times higher inhibitory activity, compared to WE. Lastly, maltase inhibition assays showed both PACF and LMWF have lower inhibitory activity compared to WE. The results suggest cranberry has an inhibitory effect against the action of alpha‐glucosidases, which is mainly due to sucrase inhibition, and is low molecular weight phenolic dependent.Grant Funding Source: This research was supported by US Army Natick Soldier Research Engineering Center