Terpenoids from Leaves of Guarea macrophylla Display In Vitro Cytotoxic Activity and Induce Apoptosis In Melanoma Cells

Planta Medica ◽  
2017 ◽  
Vol 83 (16) ◽  
pp. 1289-1296 ◽  
Author(s):  
Geanne Conserva ◽  
Natalia Girola ◽  
Carlos Figueiredo ◽  
Ricardo Azevedo ◽  
Sasha Mousdell ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Tumor Cells ◽  
Nuclear Dna ◽  
Morphological Changes ◽  
Folk Medicine ◽  
Melanoma Cells ◽  
In Vitro Assays ◽  
Apoptotic Pathway ◽  
Isolation And Characterization

Abstract Guarea macrophylla is a Brazilian plant species that has been used in folk medicine to treat a range of diseases. Our ongoing work focuses on the discovery of new bioactive natural products derived from Brazilian flora. The current study describes the identification of cytotoxic compounds from the EtOH extract of leaves from G. macrophylla using bioactivity-guided fractionation. This approach resulted in the isolation and characterization of four compounds: cycloart-23E-ene-3β,25-diol (1), (23S*,24S*)-dihydroxycicloart-25-en-3-one (2), isopimara-7,15-diene-2α,3β-diol (3), and isopimara-7,15-dien-3β-ol (4), in which 2 and 3 are identified as new derivatives. In vitro assays were conducted to evaluate the cytotoxic activity of compounds 1–4 against a panel of cancer cell lines and to determine the possible mechanism(s) related to the activity of the compounds on B16F10Nex2 cells. The most active compound 1 induced cytotoxic effects on tumor cells, with IC50 values of 18.3, 52.1, and 58.9 µM against HL-60, HeLa, and B16F10-Nex2 tumor cells, respectively. Furthermore, it was observed in melanoma cells that compound 1 induced several specific apoptotic hallmarks, such as morphological changes in the cell shape structure, nuclear DNA condensation, specific chromatin fragmentation, and disruption in the mitochondrial membrane potential, which are related to the intrinsic apoptotic pathway.

In vitro cytotoxic activity of chitosan–bullfrog oil microemulsion against melanoma cells

2015 ◽  
Vol 9 (4) ◽  
pp. 172-177 ◽  
Author(s):  
Cínthia Caetano Bonatto ◽  
Graziella Anselmo Joanitti ◽  
Luciano Paulino Silva
Keyword(s):  
Cytotoxic Activity ◽  
Melanoma Cells ◽  
Bullfrog Oil

scholarly journals Pharmacologic marrow purging in murine T cell leukemia

Blood ◽  
1988 ◽  
Vol 71 (6) ◽  
pp. 1656-1661 ◽  
Author(s):  
EA Copelan ◽  
SC Johnson ◽  
MR Grever ◽  
JF Sheridan ◽  
PJ Tutschka
Keyword(s):  
T Cells ◽  
Tumor Cells ◽  
In Vitro Assays ◽  
Cell Leukemia ◽  
Term Survival ◽  
Adenosine Deaminase Activity ◽  
Marrow Purging ◽  
Malignant T Cells

Abstract Deoxycoformycin in combination with deoxyadenosine was used to purge 6C3HED malignant T cells from murine marrow in vitro. Adenosine deaminase activity of 6C3HED cells was ablated by incubation with 10(- 6) mol/L deoxycoformycin (dCF). During a 12-hour incubation with 10(-6) mol/L dCF and 10(-4) mol/L deoxyadenosine, tumor cells sequentially accumulated dATP, became depleted of NAD followed by ATP, then died. More than 5 logs of 6C3HED cells were killed as measured by survival of mice injected with treated tumor cells. Identical incubation of 5 x 10(6) marrow cells did not interfere with rescue of syngeneic lethally irradiated mice. Long-term survival was demonstrated in 12 of 14 mice that received marrow that had been contaminated with 5% 6C3HED cells, incubated with deoxycoformycin and deoxyadenosine, then used to rescue lethally irradiated mice. This murine model provides information not available from in vitro assays and may be useful in the development of strategies to purge malignant T cells from marrow.

scholarly journals Synthesis, Structure and In Vitro Cytotoxic Activity of Novel Cinchona—Chalcone Hybrids with 1,4-Disubstituted- and 1,5-Disubstituted 1,2,3-Triazole Linkers

Molecules ◽  
2019 ◽  
Vol 24 (22) ◽  
pp. 4077 ◽  
Author(s):  
Jernei ◽  
Duró ◽  
Dembo ◽  
Lajkó ◽  
Takács ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Malignant Cell ◽  
Conformational Space ◽  
In Vitro Assays ◽  
Inhibitory Effects ◽  
Cyclization Reactions ◽  
Click Reactions ◽  
Metal Catalyzed ◽  
Dft Modelling

By means of copper(I)-and ruthenium(II)-catalyzed click reactions of quinine- and quinidine-derived alkynes with azide-substituted chalcones a systematic series of novel cinchona-chalcone hybrid compounds, containing 1,4-disubstituted- and 1,5-disubstituted 1,2,3-triazole linkers, were synthesized and evaluated for their cytotoxic activity on four human malignant cell lines (PANC-1, COLO-205, A2058 and EBC-1). In most cases, the cyclization reactions were accompanied by the transition-metal-catalyzed epimerization of the C9-stereogenic centre in the cinchona fragment. The results of the in vitro assays disclosed that all the prepared hybrids exhibit marked cytotoxicity in concentrations of low micromolar range, while the C9-epimerized model comprising quinidine- and (E)-1-(4-(3-oxo-3-(3,4,5-trimethoxyphenyl)prop-1-en-1-yl)phenyl) fragments, connected by 1,5-disubstituted 1,2,3-triazole linker, and can be regarded as the most potent lead of which activity is probably associated with a limited conformational space allowing for the adoption of a relatively rigid well-defined conformation identified by DFT modelling. The mechanism of action of this hybrid along with that of a model with markedly decreased activity were approached by comparative cell-cycle analyses in PANC-1 cells. These studies disclosed that the hybrid of enhanced antiproliferative activity exerts significantly more extensive inhibitory effects in subG1, S and G2/M phases than does the less cytotoxic counterpart.

IL-33/ST2 Axis Regulates Vasculogenic Mimicry via ERK1/2-MMP-2/9 Pathway in Melanoma

Dermatology ◽  
2019 ◽  
Vol 235 (3) ◽  
pp. 225-233 ◽  
Author(s):  
Fuhan Yang ◽  
Mingming Wen ◽  
Dayu Pan ◽  
Xian Lin ◽  
Jing Mo ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Vasculogenic Mimicry ◽  
Melanoma Cells ◽  
Tube Formation ◽  
Migration And Invasion ◽  
Rapid Progression ◽  
Inflammatory Factor ◽  
Action Methods ◽  
Significant Health

Background: Melanoma, an extremely malignant form of cancer, poses a significant health risk. Vasculogenic mimicry (VM), blood vessels formed by tumor cells instead of endothelial cells, is an important factor for the rapid progression of melanoma. Interleukin (IL)-33 is an inflammatory factor commonly found in the tumor microenvironment and plays an important role in the progression of many tumors. IL-33 acts on immune cells and tumor cells through its receptor ST2. This study hypothesized that IL-33 directly affects the progression of melanoma. Objectives: This study was designed to investigate the effect of IL-33 on VM of melanoma and its potential mechanism of action. Methods: The expression of ST2 was evaluated in 66 cases of melanoma collected from human patients, and the differences were analyzed. In vitro experiments were conducted to study the effects of the IL-33/ST2 axis on cell migration and invasion and to elucidate possible mechanisms. Results: ST2 expression is associated with that of matrix metalloproteinase (MMP)-2 and VM in melanoma of patients. IL-33 increases the abilities of proliferation, migration and invasion of melanoma cells and VM tube formation through ST2. IL-33 induces the production of MMP-2/9 via ERK1/2 phosphorylation. Conclusion: IL-33 can directly act on melanoma cells and promote its development.

scholarly journals Phosphodiesterase-1 Inhibitory Activity of Two Flavonoids Isolated fromPistacia integerrimaJ. L. Stewart Galls

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Abdur Rauf ◽  
Muhammad Saleem ◽  
Ghias Uddin ◽  
Bina S. Siddiqui ◽  
Haroon Khan ◽  
...  
Keyword(s):  
Inhibitory Activity ◽  
In Silico ◽  
Molecular Mechanisms ◽  
Methanolic Extract ◽  
Chemical Constituents ◽  
Folk Medicine ◽  
Chemical Characterization ◽  
Active Constituents ◽  
Isolation And Characterization

Pistacia integerrimais one of twenty species among the genusPistacia. Long horn-shaped galls that develop on this plant are harvested and used in Ayurveda and Indian traditional medicine to make “karkatshringi”, a herbal medicine used for the treatment of asthma and different disorders of respiratory tract. However, until now, the molecular mechanisms of action of “karkatshringi” and its chemical characterization are partially known. This study deals with the isolation and characterization of the active constituents from the methanolic extract ofP. integerrimagalls and it was also oriented to evaluatein vitroandin silicotheir potential enzymatic inhibitory activity against phosphodiesterase-1 (PDE1), a well-known enzyme involved in airway smooth muscle activity and airway inflammation. Our results showed that the methanolic extract ofP. integerrimagalls and some of its active constituents [naringenin (1) and 3,5,7,4′-tetrahydroxy-flavanone (2)] are ablein vitroto inhibit PDE1 activity (59.20 ± 4.95%, 75.90 ± 5.90%, and 65.25 ± 5.25%, resp.) and demonstratein silicoan interesting interaction with this enzymatic site. Taken together, our results add new knowledge of chemical constituents responsible for the biological activity ofP. integerrimaand contextually legitimate the use of this plant in folk medicine.

In Vitro Expansion of Blood γδ T Cells in Patients with Myeloma/Lymphoma and Anti-Tumor Cytotoxicity of the Expanded γδ T Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3895-3895
Author(s):  
Anri Saito ◽  
Miwako Narita ◽  
Norihiro Watanabe ◽  
Nozomi Tochiki ◽  
Yumi Hiroi ◽  
...  
Keyword(s):  
T Cells ◽  
Cytotoxic Activity ◽  
Tumor Cells ◽  
Γδ T Cells ◽  
Cellular Immunotherapy ◽  
Target Cells ◽  
Type I ◽  
Type I Ifn ◽  
Lymphoma Cells

Abstract In order to establish an efficient anti-tumor cellular immunotherapy using blood In order to establish an efficient anti-tumor cellular immunotherapy using blood γδ T cells, we investigated the in vitro expansion of γδ T cells in the patients with myeloma and lymphoma by the culture of PB-MNC with bisphosphonate and a low dose of IL-2 and we demonstrated the cytotoxic activity of the expanded γδ T cells against myeloma/lymphoma cells. Simultaneously we explored the potent methods for enhancing the anti-tumor cytotoxic activity of γδ T cells by both directions of activating the expanded γδ T cells and making target tumor cells sensitive to γδ T cells. For the activation of γδ T cells, expanded γδ T cells were exposed with type I IFN, monocyte-derived dendritic cells (mo-DC), or plasmacytoid dendritic cell like cell line PMDC05 (leukemia cell line established from CD4+ CD56+ acute leukemia in our laboratory) for 2 days. For the enhancement of sensitivity of target tumor cell to γδ T cells, we aimed to increase the content of IPP (the potent pyrophosphate antigen for γδ T cells) in tumor cells by decreasing the metabolic downstream of IPP. For decreasing the downstream of IPP, we tried to suppress FPP synthetase, which is involved in downstream metabolism of IPP, by using nitrogen-containing bisphosphonate. In addition, the expression of stress-induced molecules such as MICA/B on target tumor cells was evaluated in association with the level of cytotoxicity of γδ T cells against the tumor cells. Compared with normal control, the patients with myeloma (n=8) demonstrated decreased percentage and counts of PB γδ T cells. Patients with lymphoma (n=7) showed a wide range of values in PB γδ T cells, covering a normal range. Amplification rate of PB γδ T cells by culture with zoledronate and IL-2 varied markedly from patient to patient up to 120 times in myeloma and 90 times in lymphoma. Expanded γδ T cells generated in patients with myeloma/lymphoma were demonstrated to possess the cytotoxic activity against myeloma/lymphoma cells by 51Cr-release assay and CFSE-labeled target cell. The cytotoxic activity of expanded γδ T cells was enhanced by the exposure of γδ T cells with type I IFN (IFN-α and IFN-β). The activation of γδ T cells, which was evaluated by the elevation of CD69 expression, was observed by the exposure of γδ T cells with type I IFN, mo-DC, or PMDC05 for 2 days. The sensitivity of target myeloma/lymphoma cells to γδ T cells was enhanced by the exposure of the target cells to bisphosphonate such as zoledronate. The expression level of MICA/B on target tumor cells was demonstrated to be associated with the potency of cytotoxicity of γδ T cells against the tumor cells. The present study demonstrated that γδ T cells expanded from myeloma/lymphoma patient’s blood are cytotoxic to myeloma/lymphoma cells. There are two methods practically available for enhancing the cytotoxic activity of expanded γδ T cells against myeloma/lymphoma cells, one of which is activating γδ T cells and the other is elevating the sensitivity of target cells by using bisphosphonate.

scholarly journals RBP-L, a transcription factor related to RBP-Jkappa.

1997 ◽  
Vol 17 (5) ◽  
pp. 2679-2687 ◽  
Author(s):  
S Minoguchi ◽  
Y Taniguchi ◽  
H Kato ◽  
T Okazaki ◽  
L J Strobl ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Notch Receptor ◽  
In Vitro Assays ◽  
Functional Interaction ◽  
Protein Protein Interaction ◽  
Isolation And Characterization ◽  
Signalling Molecule

RBP-Jkappa is a sequence-specific DNA binding protein which plays a central role in signalling downstream of the Notch receptor by physically interacting with its intracellular region. Although at least four Notch genes exist in mammals, it is unknown whether each Notch requires a specific downstream signalling molecule. Here we report isolation and characterization of a mouse RBP-Jkappa-related gene named RBP-L that is expressed almost exclusively in lung, in contrast to the ubiquitous expression of RBP-Jkappa. For simplicity, we propose to call RBP-Jkappa RBP-J. The RBP-L protein bound to a DNA sequence almost identical to that of RBP-J. Surprisingly, RBP-L did not interact with any of the known four mouse Notch proteins. Although we found that RBP-L and EBNA-2 cooperated in transcriptional activation, they did not show significantly strong protein-protein interaction that can be detected by several in vivo and in vitro assays. This is again in contrast to physical association of RBP-J with EBNA-2. Several models to explain functional interaction between RBP-L and EBNA-2 are discussed.

scholarly journals Evaluation of the cytotoxic potential of extracts from the genus Passiflora cultived in Brazil against cancer cells

2018 ◽  
Author(s):  
Ricardo Guimarães Amaral ◽  
Silvana Vieira Floresta Gomes ◽  
Ângelo Roberto Antoniolli ◽  
Maria Claudia dos Santos Luciano ◽  
Cláudia do Ó Pessoa ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Cytotoxic Activity ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Cytotoxic Potential ◽  
Leaf Extracts ◽  
High Cytotoxic Activity ◽  
Apoptosis And Necrosis

AbstractThis work aimed to evaluate the cytotoxic potential against cancer cells of Passiflora genus plant species cultivated in Brazil and identify the mechanism of cytotoxicity induced by the most promising extract. Leaf extracts from 14 Passiflora (P.) species were obtained ASE and in vitro cytotoxicity evaluated against cancer cell lines using MTT assay at a single concentration of 50 μg/mL. Additionally, the IC50 of the P. alata (ELPA) leaf extracts was determined against both tumor (HCT-116, SF-295, OVACAR-8, and HL-60), and non-tumor cells (PBMC). The ELPA flavonoids were identified by HPLC-DAD and UHPLC-MS/MS. The morphological analyses used light and fluorescence microscopy, and cell cycle and DNA fragmentation analyses used flow cytometry to determine the mechanism of cell death induced by ELPA in HL-60. Among the Passiflora leaf extracts evaluated; ELPA stood out with high cytotoxic activity, followed by P. capsularis and P. quadrangulares with varying high and low cytotoxic activity. ELPA presented high cytotoxic potency in HL-60 (IC50 19.37 μg/mL), yet without cytotoxic activity against PBMC, suggesting selectivity for tumor cells. The cytotoxic activity of ELPA may well be linked to the presence of ten identified flavonoids. Cells treated with ELPA presented the hallmarks typical of apoptosis and necrosis, with cell cycle arrest in the G2/M phase. Conclusion: From among the studied species, ELPA presented greater cytotoxic activity, possibly a consequence of synergistic flavonoid action which induces cell death by apoptosis and necrosis.

Growth Inhibition of Murine Colonic Adenocarcinoma by Tumor Immune but not by IL-2-Activated or Alloactivated Lymphocytes

1987 ◽  
Vol 73 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Monica Rodolfo ◽  
Giorgio Parmiani
Keyword(s):  
Tumor Growth ◽  
Tumor Cells ◽  
Interleukin 2 ◽  
In Vitro Assays ◽  
Growing Cell ◽  
Spleen Lymphocytes ◽  
Antigenic Profile ◽  
Colonic Adenocarcinomas

The antigenic profile of C-26 and C-51 BALB/c colonic adenocarcinomas was examined by in vivo and in vitro assays. Mice immunized with irradiated C-26 or C-51 tumor cells from freshly excised tumor nodules or from in vitro-growing cell lines were able to reject a challenge of both tumors. Spleen lymphocytes of immune but not of normal mice were effective in cross-inhibiting tumor growth in vivo in a Winn assay. Tissue-associated antigens common to C-26 and C-51 and to their metastases but not to other syngeneic neoplasms were detected in vitro by cytotoxic T lymphocytes obtained after 5 days of a secondary culture of immune lymphocytes and irradiated tumor cells. Activated lymphocytes were obtained by exposure of spleen cells to interleukin 2 or by allostimulation. Such lymphocytes, although cytotoxic in vitro on C-26 and C-51 carcinomas, were unable to significantly reduce in vivo tumor growth in the Winn assay.

BXQ-350 to target to the lysosome and kill glioblastoma (GBM) cells via activation of apoptotic caspases in vitro.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. 3639-3639
Author(s):  
Laura Felix ◽  
Timothy J Stephens ◽  
Nikhil J Wilkins
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Tumor Cells ◽  
Lysosomal Membrane ◽  
Sphingolipid Metabolism ◽  
Activity Levels ◽  
Caspase 9 ◽  
Apoptotic Pathway ◽  
Caspase 7

3639 Background: Apoptosis is a programmed cell death mechanism where cells respond to internal or external stimuli by initiating a cascade of events and enzymes leading to cell death. One of the hallmarks of cancer is the ability of tumor cells to resist these apoptotic stimuli. This allows tumor cells to have aberrant metabolisms, such as sphingolipid metabolism in tumor cell lysosomes, or mutations which would normally commit cells to death. Saposin C, the protein component of BXQ-350, Bexion Pharmaceuticals’ proprietary biotherapeutic, is involved in normal lysosomal sphingolipid metabolism. Removing resistance, shortcutting steps leading to apoptosis, or correcting sphingolipid metabolism can result in the death of these tumor cells. Methods: The GBM cell line Gli36ΔEGFR was plated in 96 well plates at a density of 1x104 cells per well in Dulbecco’s Modified Eagle Media with 10% FBS overnight at 37oC for caspase and cytotoxicity assays. Cells were treated with 9uM to 30uM BXQ-350 in triplicate and incubated for 24 hours at 37oC. Promega’s Caspase-Glo 9 or Caspase-Glo 3/7 reagent was added to appropriate wells and the plates were incubated at room temperature in the dark for 3 hours then luminescence was read. The parallel cytotoxic assay was run under the same conditions except Roche’s MTT labeling reagent was added to the appropriate wells after 24 hours and incubated at 370C for 4 hours. Solubilization solution was added to each well and the plate was incubated at 37oC overnight then absorbance was read. The GBM cell line U87 MG was used to determine lysosomal targeting. U87 MG cells were treated with 10uM BXQ-350 and incubated at 37oC overnight. They were stained with anti-SapC (RFP) and anti-LAMP1 (GFP) antibodies and images were taken. Results: BXQ-350 mediated cell death is correlated with a rise in Caspase 3, Caspase 7 and Caspase 9 activity. The caspase activity levels did not rise until after BXQ-350 passed its IC50 and stayed elevated. Caspases 3/7 levels showed higher activity compared to untreated than Caspase 9. In addition to this, BXQ-350 was seen to colocalize to LAMP1, a lysosomal membrane protein. Conclusions: BXQ-350 tracks to the lysosomal membrane where it initiates the cascade of enzymes necessary to cause apoptosis. Caspases 3/7 are the effector caspases and are necessary for the completion of the apoptotic pathway. The higher activity levels of these caspases show the cells are committed to cell death not allowing these cells to subvert apoptosis. This removes one of the major barriers to fighting cancer.

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