Dedifferentiation of leaf explants and antileukemia activity of an ethanolic extract of cell cultures of Moringa oleifera

The present study was aimed at developing an efficient protocol for callus induction from the leaves of Moringa oleifera and to investigate its crude extract antileukemia activity on leukemia cells. Several secondary metabolites are present in M. oleifera as the plant serves as reservoirs for various bioactive compounds. Callus cultures of M. oleifera were induced from leaf explants incubated on MS medium supplemented with different concentrations of 2,4-dichloro-phenoxyacetic acid (2,4-D). The crude extracts of the callus were evaluated in vitro for their activity against leukemia cells and hepatocarcinoma. Among the different concentrations, 2,4-D at 0.1 mg/l induced highest frequencies of callus growth index (7.8) when compared with other concentrations. Ethanolic extracts killed about 36% of abnormal cells among primary cells harvested from 3 patients with acute myeloid leukemia (AML) and hepatocarcinoma cells HpG2. These results provide an in vitro evidence and support the traditional use of M. oleifera leaf as a potent source of anticancer. However, more researches are needed at phytochemical and clinical levels to confirm the traditional use of this plant as anticancer.Keywords: Moinga olifera, callus culture, antileukemia, hepatocarcinom

Willow Leaves' Extracts Contain Anti-Tumor Agents Effective against Three Cell Types

Many higher plants contain novel metabolites with antimicrobial, antifungal and antiviral properties. However, in the developed world almost all clinically used chemotherapeutics have been produced by in vitro chemical synthesis. Exceptions, like taxol and vincristine, were structurally complex metabolites that were difficult to synthesize in vitro. Many non-natural, synthetic drugs cause severe side effects that were not acceptable except as treatments of last resort for terminal diseases such as cancer. The metabolites discovered in medicinal plants may avoid the side effect of synthetic drugs, because they must accumulate within living cells. The aim here was to test an aqueous extract from the young developing leaves of willow (Salix safsaf, Salicaceae) trees for activity against human carcinoma cells in vivo and in vitro. In vivo Ehrlich Ascites Carcinoma Cells (EACC) were injected into the intraperitoneal cavity of mice. The willow extract was fed via stomach tube. The (EACC) derived tumor growth was reduced by the willow extract and death was delayed (for 35 days). In vitro the willow extract could kill the majority (75%–80%) of abnormal cells among primary cells harvested from seven patients with acute lymphoblastic leukemia (ALL) and 13 with AML (acute myeloid leukemia). DNA fragmentation patterns within treated cells inferred targeted cell death by apoptosis had occurred. The metabolites within the willow extract may act as tumor inhibitors that promote apoptosis, cause DNA damage, and affect cell membranes and/or denature proteins

Figure 1

<p>The effect of willow young leaves extracts on the viability of ALL cells. 7 ALL patients (acute lymphoblastic leukemia) 0.1 ml of the cell suspension containing 10⁵ cells was added to 5 tubes. To three of the tubes, 0.1 ml of the willow extract was added, while the other two tubes served as negative and positive controls. Culture medium was used instead of the willow extract for the negative control and the willow extract was added to the cells from healthy volunteers as a positive control. The tubes were incubated for 24 h and tested for their viability using the trypan blue.</p

Figure 5

<p>The mode of action of the Salicin and Saligenin to destroy immature White Blood Cells (WBC's) in leukemia patients.</p

Figure 3

<p>The effect of willow young leaves extracts on the viability of Ehrlich asites carcinoma cells (EACC). 0.1 ml of the cell suspension containing 10⁵ cells was added to 4 tubes. To three of the tubes, 0.1 ml of the willow extract was added, while the other tube served as negative control. Culture medium was used instead of the willow extract for the negative control. The tubes were incubated for 24 h and tested for their viability using the trypan blue.</p

Figure 4

<p>Effect of salix water extract on surviving of transplanted animals (EACC). One group fed on normal diet (negative control), second group was fed on normal diet and transplanted to the intraperitoneal (i.p) cavity with EACC at 2×10⁶ cells (0.2 ml) (positive control), third group fed the normal diet and EACC infected then each mouse daily forced to ingest orally via stomach tube about 0.2 ml of an aqueous willow extract (10% w/v), fourth group were treated daily like the third group except with 0.6 ml willow extract.</p

Figure 2

<p>The effect of willow young leaves extracts on the viability of AML cells. 13 AML patients (acute myeloid leukemia) 0.1 ml of the cell suspension containing 10⁵ cells was added to 5 tubes. To three of the tubes, 0.1 ml of the willow extract was added, while the other two tubes served as negative and positive controls. Culture medium was used instead of the willow extract for the negative control and the willow extract was added to the cells from healthy volunteers as a positive control. The tubes were incubated at tested for their viability using the trypan blue.</p

Predicting In Silico Which Mixtures of the Natural Products of Plants Might Most Effectively Kill Human Leukemia Cells?

The aim of the analysis of just 13 natural products of plants was to predict the most likely effective artificial mixtures of 2-3 most effective natural products on leukemia cells from over 364 possible mixtures. The natural product selected included resveratrol, honokiol, chrysin, limonene, cholecalciferol, cerulenin, aloe emodin, and salicin and had over 600 potential protein targets. Target profiling used the Ontomine set of tools for literature searches of potential binding proteins, binding constant predictions, binding site predictions, and pathway network pattern analysis. The analyses indicated that 6 of the 13 natural products predicted binding proteins which were important targets for established cancer treatments. Improvements in effectiveness were predicted for artificial combinations of 2 or 3 natural products. That effect might be attributed to drug synergism rather than increased numbers of binding proteins bound (dose effects). Among natural products, the combinations of aloe emodin with mevinolin and honokiol were predicted to be the most effective combination for AML-related predicted binding proteins. Therefore, plant extracts may in future provide more effective medicines than the single purified natural products of modern medicine, in some cases

Active principle from Moringa oleifera Lam leaves effective against two leukemias and a hepatocarcinoma

Medicinal plants are important elements of indigenous medical system that have persisted in developing countries. Many of the pharmacological principles currently used as anticancer agents were first isolated from plants. However, some important anticancer agents are still extracted from plants because they cannot be synthesized chemically on a commercial scale due to their complex structures that often contain several chiral centers. The aim of this study was to test different extracts from the leaves of Moringa or drumstick tree (Moringa oleifera) for activity against leukemia and hepatocarcinoma cells in vitro. The extracts could kill majority (70 - 86%) of the abnormal cells among primary cells harvested from 10 patients with acute lymphoblastic leukemia (ALL) and 15 with acute myeloid leukemia (AML) as well as a culture of hepatocarcinoma cells (75% death), but most significantly by the hot water and ethanol extracts. In conclusion, M. oleifera may have potential for use as source of natural treatment for diseases such as cancer.Key words: Moringa oleifera, anti-cancer, acute lymphoblastic leukemia, acute myeloid leukemia, hepatocarcinoma

Biochemical and molecular investigation of in vitro antioxidant and anticancer activity spectrum of crude extracts of willow leaves salix safsaf

Organic fractions and extracts of willow (Salix safsaf) leaves, produced by sequential solvent extraction as well as infusion and decoction, exhibited anticancer potencies in four cancerous cell lines, including breast (MCF-7), colorectal (HCT-116), cervical (HeLa) and liver (HepG2). Results of the MTT assay revealed that chloroform (CHCl3) and ethyl acetate (EtOAc)-soluble fractions exhibited specific anticancer activities as marginal toxicities were observed against two non-cancerous control cell lines (BJ-1 and MCF-12). Ultra-high-resolution mass spectrometry Q-Exactive™ HF Hybrid Quadrupole-Orbitrap™ coupled with liquid chromatography (UHPLC) indicated that both extracts are enriched in features belonging to major phenolic and purine derivatives. Fluorescence-activated cell sorter analysis (FACS), employing annexin V-FITC/PI double staining indicated that the observed cytotoxic potency was mediated via apoptosis. FACS analysis, monitoring the increase in fluorescence signal, associated with oxidation of DCFH to DCF, indicated that the mechanism of apoptosis is independent of reactive oxygen species (ROS). Results of immunoblotting and RT-qPCR assays showed that treatment with organic fractions under investigation resulted in significant up-regulation of pro-apoptotic protein and mRNA markers for Caspase-3, p53 and Bax, whereas it resulted in a significant reduction in amounts of both protein and mRNA of the anti-apoptotic marker Bcl-2. FACS analysis also indicated that pre-treatment and co-treatment of human amniotic epithelial (WISH) cells exposed to the ROS H2O2 with EtOAc fraction provide a cytoprotective and antioxidant capacity against generated oxidative stress. In conclusion, our findings highlight the importance of natural phenolic and flavonoid compounds with unparalleled and unique antioxidant and anticancer properties