BackgroundDendritic cells (DCs) are unique antigen presenting cells (APC) which play a pivotal role in immunotherapy and induction of an effective immune response against tumors. In the present study, 80% ethanol extract of Phyllanthus amarus was used to generate tumor lysate (TLY) derived from HCT 116 and MCF-7 cancer cell lines via induction of apoptosis. Monocyte-derived DCs were generated ex vivo from the adherent population of peripheral blood mononuclear cells (PBMCs). The generated TLY were used to impulse DCs to investigate its effect on their cellular immune functions including antigen presentation capacity, phagocytic activity, chemotaxis capacity, T-cell proliferation and cytokines release.MethodsThe effect of P. amarus-generated TLY on DCs maturation was evaluated by determination of MHC class I, II and CD 11c expression as well as the co-stimulatory molecules CD 83 and 86 by using flow cytometry. The phagocytic capacity of TLY-pulsed DCs was investigated through FITC-dextran uptake by using flow cytometry. The effect on the cytokines release including IL-12, IL-6 and IL-10 was elucidated by using ELISA. The migration capacity and T cell proliferation activity of pulsed DCs were measured. The relative gene expression levels of cytokines were determined by using qRT-PCR. The major constituents of P. amarus extract were qualitatively and quantitatively analyzed by using validated reversed-phase high performance liquid chromatography (HPLC) methods.ResultsP. amarus-generated TLY significantly up-regulated the expression levels of MHC class I, CD 11 c, CD 83 and 86 in pulsed DCs. The release of interleukin IL-12 and IL-6 was enhanced by TLY-DCs at a ratio of 1 DC: 3 tumor apoptotic bodies (APO), however, the release of IL-10 was suppressed. The migration ability as well as allogeneic T-cell proliferation activities of loaded DCs were significantly enhanced, but their phagocytic capacity was highly attenuated. The gene expression profiles for IL-12 and IL-6 of DCs showed increase in their mRNA gene expression in TLY pulsed DCs versus unloaded and LPS-treated only DCs.ConclusionThe effect of P. amarus-generated TLY on the immune effector mechanisms of DCs verified its potential to induce an in vitro anti-tumor immune response against the recognized tumor antigen.

Background Seagrasses are unique marine flowering plants. Enhalus acoroides (L.f.) Royle (family Hydrocharitaceae), a new record for the Egyptian coast of the Red Sea, was the grass of choice. A comparative study on Enhalus acoroides (L.f.) Royle (leaves, roots and rhizomes) was done to determine the plant organ that shows the highest antiproliferative and antioxidant activities. The total phenolic content was determined using the Folin–Ciocalteu method. The total flavonoid content was estimated by the aluminum chloride assay. Fourier transform infrared (FT-IR) analysis was performed to detect the chemical functional groups in the extract. High-performance liquid chromatography (HPLC) was done for the qualitative and quantitative evaluation of phenolic compounds. UPLC-ESI-TOF–MS was performed for metabolomics profiling of the extract. Antioxidant activity was determined using the DPPH scavenger percentage method. Antiproliferation assay against hepatocellular carcinoma HepG2, human breast cancer cell lines MCF-7, MDA-MB-231 was performed for the three seagrass organs. Mitochondrial membrane potential (ΔΨm) was measured after treatment with three extracts against MCF-7 cell line. Results The highest phenolic content is found in the leaves, while roots exhibited the highest DPPH scavenger percentage. The total concentration of phenolic compounds detected by HPLC was leaves > rhizomes > roots. Also leaves exhibit the highest antiproliferative activity and mitochondrial membrane potential depletion effect against MCF-7 cell line tested. UPLC-ESI-TOF–MS metabolite profiling of leaves detected different secondary and primary metabolites to which the activity was retained. Leaves are a new candidate to be used in the treatment of cancer. Conclusion Enhalus acoroides (L.f.) Royle leaves extract is a new nutraceutical candidate. Further in-depth studies are required on Enhalus acoroides (L.f.) Royle leaves. Graphical abstract

Background: The anticancer effects of Phyllanthus amarus extract on various cancer cells have been investigated, however, effect of its major constituents on HCT116 human colorectal cancer cells has not been reported. Objective: In the present study, we investigated the cytotoxic effect of 80% ethanol extract of P. amarus and its marker constituents (phyllanthin, hypophyllanthin, gallic acid, niranthin, greraniin, phyltetralin, isolintetralin, corilagin and ellagic acid) on HCT116 and their underlying mechanisms of action. Method: Their anti-proliferative and apoptotic effects on HCT 116 were performed using MTT assay and flow cytometric analysis, respectively, while caspases 3/7, 8 and 9 activities were examined using colorimetric method. The expression of cleaved poly ADP ribose polymerase enzyme (PARP) and cytochrome c proteins was investigated by immune-blot technique. Results: HPLC and LC-MS/MS analyses demonstrated the extract contained mainly lignans and polyphenols. The plant samples markedly suppressed the growth and expansion of HCT116 cells concentration- and time-dependently with no toxicity against normal human fibroblast CCD18 Co. P. amarus extract, phyllanthin and gallic acid induced mode of cell death primarily via apoptosis as confirmed by the exteriorization of phosphatidylserine. Caspases 3/7, 8, and 9 activities increased concentration-dependently following 24 h treatment. The expressions of cleaved PARP (Asp 214) and cytochrome c were markedly upregulated. Conclusion: P. amarus extract, phyllanthin and gallic acid exhibited apoptotic effect on HCT116 cells via caspases-dependent pathway.