scholarly journals Study on the Extraction and Antitumor Activity of Achyranthes Bidentata Polysaccharide

2018 ◽  
Vol 10 (2) ◽  
pp. 23
Author(s):  
Linsong Yang ◽  
Yuhuan Shen ◽  
Fang Wang
Keyword(s):  
Therapeutic Agent ◽  
Annexin V ◽  
Extraction Temperature ◽  
Double Staining ◽  
Dna Ladder ◽  
Achyranthes Bidentata ◽  
Mtt Method ◽  
Anti Tumor Activity ◽  
Optimum Extraction ◽  
Yield And Purity

Achyranthes bidentata polysaccharides (ABPS), a major active ingredient of Achyranthes bidentata, has many kinds of well-known pharmacological activity, such as anti-coagulation, indirect anti-tumor, anti-aging and hypoglycemic effect, and also have certain effects on the immune system. However, the direct anti-tumor activity of ABPS remain unclear. In this paper, ABPS was efficiently isolated from Achyranthes bidentata adopted orthogonal experimental method, and then its anti-tumor effect on LTEP-a-2 cells was investigated. LTEP-a-2 cells were treated with various concentrations of ABPS (0, 0.2, 0.4, 0.6, 0.8 and 1 mg/mL) for 48h. Subsequently, cell viability was analyzed by MTT method and the effects of ABPS on the apoptosis of LTEP-a-2 cells was explored by DNA ladder, PI and Annexin V FITC/PI double staining. The results showed that the optimum extraction conditions of ABPS was that the extraction temperature at 90oC, 1:40(W/V) of solid to liquid, and extraction time 4 h. In addition, the yield and purity of ABP was measured up to 15.52% and 97.21% respectively. Furthermore, ABPS significantly inhibited the proliferation and induced the apoptosis of LTEP-a-2 cells. So ABPS might be utilized as a new potential therapeutic agent against Lung Adenocarcinoma.

Derivatives of Deoxypodophyllotoxin Induce Apoptosis Through Bcl-2/Bax Proteins Expression

2020 ◽  
Vol 20 ◽  
Author(s):  
Ya-Nan Li ◽  
Ni Ning ◽  
Lei Song ◽  
Yun Geng ◽  
Jun-Ting Fan ◽  
...  
Keyword(s):  
Annexin V ◽  
Mtt Method ◽  
Anthriscus Sylvestris ◽  
Anti Tumor Activity ◽  
Derivatives Of ◽  
Toxicity In Vitro ◽  
Ht 29 ◽  
Mcf 7

Background: Deoxypodophyllotoxin, isolated from theTraditional Chinese Medicine Anthriscus sylvestris, is well-known because of its significant antitumor activity with strong toxicity in vitro and in vivo. Objective: In this article, we synthesized a series of deoxypodophyllotoxin derivatives, and evaluated their antitumor effectiveness.Methods:The anti tumor activity of deoxypodophyllotoxin derivatives was investigated by the MTT method. Apoptosis percentage was measured by flow cytometer analysis using Annexin-V-FITC. Results: The derivatives revealed obvious cytotoxicity in the MTT assay by decreasing the number of late cancer cells. The decrease of Bcl-2/Bax could be observed in MCF-7, HepG2, HT-29 andMG-63 using Annexin V-FITC. The ratio of Bcl-2/Bax in the administration group was decreased, which was determined by the ELISA kit. Conclusion: The derivatives of deoxypodophyllotoxin could induce apoptosis in tumor cell lines by influencing Bcl-2/Bax.

Pre-clinical evaluation of PV-10 for in vitro anti-tumor activity in refractory and high-risk adult solid tumors.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e14544-e14544
Author(s):  
Son Tran ◽  
Satbir Thakur ◽  
Mohit Jain ◽  
Chunfen Zhang ◽  
Aru Narendran
Keyword(s):  
High Risk ◽  
Tumor Cell ◽  
Solid Tumors ◽  
Cell Lines ◽  
Therapeutic Agent ◽  
Intratumoral Injection ◽  
Tumor Cell Lines ◽  
Clinical Testing ◽  
Anti Tumor Activity

e14544 Background: PV-10 (10% rose bengal disodium; 4,5,6,7-tetrachloro-2’,4’,5’,7’-tetraiodofluorescein) is a novel therapeutic agent previously shown to have potent anti-tumor activity following intratumoral injection in melanoma and refractory neuroblastoma, and currently is undergoing clinical testing as a single-agent for refractory metastatic neuroendocrine cancer (NCT02693067) and in combination with checkpoint inhibitors for metastatic melanoma (NCT02557321) and metastatic uveal melanoma (NCT00986661). Given the established clinical efficacy of PV-10 in adult melanoma and hepatic cancers via intratumoral injection, there is a need to evaluate the therapeutic potential of PV-10 in high-risk and refractory adult solid tumors via systemic administration. Our study aims to identify the clinical potential of systemically-delivered PV-10 by first generating prerequisite in vitro data for adult malignancies. Methods: Cytotoxicity assays were performed using the Alamar Blue assay to study the effects of PV-10 in vitro 96-hours post-treatment against a panel of adult solid tumor cell lines derived from breast (MCF-7, T-47D, MDA-MB-231), colorectal (HCT-116, LoVo, T-84), head and neck (CAL-27, Detroit-562, FaDu, UM-SCC-1), and testicular (NCC-IT, NTERA-2, TCAM-2) tissues. Light microscopy and Western blotting were used to investigate apoptosis induction and target modulation in tumor cells after PV-10 treatment. Results: In vitro results from our study demonstrate that PV-10 is cytotoxic at pharmacologically relevant concentrations across the indicated cell lines. Specifically, tumor cell lines originating from testicular tissues were highly sensitive to PV-10 treatment (Mean ± SD IC50: 37.5 ± 16.4 µM; n = 3) compared to breast (117.5 ± 71.0 µM; n = 3), colorectal (64.79 µM; n = 3), and head and neck (106.6 ± 29.2 µM; n = 4) cell lines. Western blot analyses showed dose- and time-dependent activation of pro-apoptotic protein markers in caspase-3 and PARP cleavage, indicating drug-induced apoptosis. Conclusions: This study provides the first pre-clinical results of PV-10 as a novel systemically-delivered therapeutic agent for a range of high-risk and refractory adult solid tumors. Data obtained from our in vitro experiments using a broad repertoire of cell lines that represent diverse molecular and phenotypic subtypes of solid tumors in adults can serve as prerequisite pre-clinical data to establish clinical testing in these populations.

scholarly journals Analysis of programmed cell death in mouse fetal oocytes

Reproduction ◽  
2007 ◽  
Vol 134 (2) ◽  
pp. 241-252 ◽  
Author(s):  
A M Lobascio ◽  
F G Klinger ◽  
M L Scaldaferri ◽  
D Farini ◽  
M De Felici
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Specific Inhibitor ◽  
Annexin V ◽  
Death Process ◽  
Parp Cleavage ◽  
Tunel Staining ◽  
Calpain Inhibitor ◽  
Dna Ladder ◽  
Caspase 2

We report a short-term culture system that allowsto define novel characteristic of programmed cell death (PCD) in fetal oocytes and to underscore newaspects of this process. Mouse fetal oocytes culturedin conditions allowingmeiotic prophase I progression underwent apoptotic degeneration waves as revealed by TUNEL staining. TEM observations revealed recurrent atypical apoptotic morphologies characterized by the absence of chromatin margination and nuclear fragmentation; oocytes with autophagic and necrotic features were also observed. Further characterization of oocyte death evidenced DNA ladder, Annexin V binding, PARP cleavage, and usually caspase activation (namely caspase-2). In the aim to modulate the oocyte death process, we found that the addition to the culture medium of the pancaspase inhibitors Z-VAD orcaspase-2-specific inhibitor Z-VDVAD resulted in a partial and transient prevention of this process. Oocyte death was significantly reduced by the antioxidant agent NAC and partly prevented by KL and IGF-I growth factors. Finally, oocyte apoptosis was reduced by calpain inhibitor I and increased by rapamycin after prolonged culture.These results support the notion that fetal oocytes undergo degeneration mostly by apoptosis. This process is, however, often morphologically atypical and encompasses other forms of cell death including caspase-independent apoptosis and autophagia. The observation that oocyte death occurs mainly at certain stages of meiosis and can only be attenuated by typical anti-apoptotic treatments favors the notion that it is controlled at least in part by stage-specific oocyte-autonomous meiotic checkpoints and when activated is little amenable to inhibition being the oocyte able to switch back and forth among different death pathways.

scholarly journals Ultrasound-Assisted Deep Eutectic Solvent Extraction of Anthocyanins from Blueberry Wine Residues: Optimization, Identification, and HepG2 Antitumor Activity

Molecules ◽  
2020 ◽  
Vol 25 (22) ◽  
pp. 5456
Author(s):  
Hongkun Xue ◽  
Jiaqi Tan ◽  
Qian Li ◽  
Jintian Tang ◽  
Xu Cai
Keyword(s):  
Solvent Extraction ◽  
Antitumor Activity ◽  
Deep Eutectic Solvent ◽  
Extraction Process ◽  
Ultrasonic Power ◽  
Extraction Temperature ◽  
Brewing Process ◽  
Ultrasound Assisted ◽  
Optimum Extraction ◽  
Better Than

Blueberry wine residues produced during the wine-brewing process contain abundant anthocyanins and other bioactive compounds. To extract anthocyanins from blueberry wine residues more efficiently, a novel procedure of ultrasound-assisted deep eutectic solvent extraction (UADESE) was proposed in this work. The extraction process was optimized by response surface methodology coupled with genetic algorithm. The optimum extraction parameters to achieve the highest yield of anthocyanins (9.32 ± 0.08 mg/g) from blueberry wine residues by UADESE were obtained at water content of 29%, ultrasonic power of 380 W, extraction temperature of 55 °C, and extraction time of 40 min. The AB-8 macroporous resin combined with Sephadex LH-20 techniques was used to purify the crude extract (CE) obtained under optimum extraction conditions and analyze the anthocyanins composition by HPLC-ESI-MS/MS. The cyanidin-3-rutinoside with purity of 92.81% was obtained. The HepG2 antitumor activity of CE was better than that of the purified anthocyanins component. Moreover, CE could increase the intracellular reactive oxygen species levels and the apoptosis, and arrest HepG2 cells in the S phases. These findings provided an effective and feasible method for anthocyanins extraction, and reduced the environmental burden of this waste.

9-Methoxycamptothecin from Nothapodytes foetida Induces Apoptosis in Murine Sarcoma S180 Cells

2011 ◽  
Vol 66 (9-10) ◽  
pp. 471-476
Author(s):  
Na Liao ◽  
Peng Zhang ◽  
Mingzhang Ao ◽  
Jing Wang ◽  
Yueyuan Shi ◽  
...  
Keyword(s):  
Cell Death ◽  
Nmr Spectroscopy ◽  
Mtt Assay ◽  
Propidium Iodide ◽  
Annexin V ◽  
Double Staining ◽  
Nothapodytes Foetida ◽  
Ic50 Value ◽  
Mitochondria Pathway ◽  
Murine Sarcoma

9-Methoxycamptothecin (MCPT) was found to have antitumour activities through topoisomerase inhibition. However, the type of cell death induced in the tumour cells treated with MCPT was not elucidated. In this study, MCPT and camptothecin were isolated from Nothapodytes foetida distributed in Hubei Province, China and identifi ed by NMR spectroscopy. MCPT was tested by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay using camptothecin as reference. Annexin V-FITC/propidium iodide double staining and real-time PCR were also performed. The IC50 value was (0.385 ± 0.08) μM. The apoptosis rates increased from 9.5% to 17.27%, 30.14%, and 66.46% with an increase in MCPT concentrations from 0, 0.19, 0.38, to 0.95 μM, respectively. The ratio of Bax/Bcl-2 also increased from 1 to 1.61, 2.43, and 4.57, respectively. Bax and Bcl-2 are crucial to the mitochondria pathway. The results indicate that the mitochondria pathway may be involed in MCPT-induced murine sarcoma S180 apoptosis.

Imatinib Induces Apoptosis of Myeloma Cells by Inhibiting c-Kit Signaling Transduction.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4738-4738
Author(s):  
Juan Li ◽  
Beihui Huang ◽  
Ying Zhao ◽  
Shaokai Luo
Keyword(s):  
Annexin V ◽  
Growth Index ◽  
Substantial Effect ◽  
Phase Cell ◽  
Imatinib Treatment ◽  
Dependent Manner ◽  
Dna Ladder ◽  
Xtt Assay ◽  
Signaling Transduction ◽  
Myeloma Cells

Abstract Objective To evaluate the mechanism and influence of Imatinib on myeloma cells which express c-Kit receptor. Methods KM3 cells were treated with Imatinib for indicated time and indicated concentration and cell growth index were evaluated by XTT assay. Cell cycle were examined by flow cytometry. Apoptosis were assessed by Annexin V/PI. c-Kit and phosphate c-Kit were examined by Western blot. Results Imatinib inhibited proliferation of KM3 cells in a concentration more than 0.25μmol/L, induced a progressive decline in S phase cell fraction and an increase in G0/G1 cells. Annexin V/PI staining and DNA ladder indicated that Imatinib had a substantial effect on apoptosis of KM3 cells in a dose-dependent manner and induced pro-caspase-3 and PARP to cleave. Imatinib treatment inhibited expression of c-Kit and provoked a decreased of IL-6 induced c-Kit phosphorylation. Conclusion Imatinib inhibites KM3 myeloma cells proliferation and induces apoptosis of KM3 myeloma cells by inhibiting c-Kit signaling transduction.

NPI-2358, a Novel Vascular Disrupting Agent Blocks Growth and Angiogenesis in Multiple Myeloma Cells.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3842-3842
Author(s):  
Dharminder Chauhan ◽  
Ajita V. Singh ◽  
Madhavi Bandi ◽  
Noopur Raje ◽  
Robert L Schlossman ◽  
...  
Keyword(s):  
Cell Lines ◽  
Annexin V ◽  
Vascular Endothelial ◽  
Caspase 9 ◽  
Migration Assays ◽  
Induced Apoptosis ◽  
Rpmi 8226 ◽  
Anti Tumor Activity

Abstract Abstract 3842 Poster Board III-778 Background and Rationale Vascular disrupting agents (VDAs) act via selectively disrupting established tumor vasculature and have shown remarkable clinical success as anti-cancer therapies. NPI-2358 is a novel VDA with a distinct structure and mechanism of action from other available VDAs. NPI-2358 binds to the colchicine-binding site of beta-tubulin preventing polymerization and disrupting the cytoplasmic microtubule network, thereby causing loss of vascular endothelial cytoskeletal function, and inducing cytotoxicity in cancer cells. Here, we examined the anti-angiogenic and anti-tumor activity of NPI-2358 in multiple myeloma (MM) cells using both in vitro and in vivo model systems. Material and Methods We utilized MM.1S, MM.1R, RPMI-8226, U266, and INA-6 human MM cell lines, as well as purified tumor cells from MM patients relapsing after prior anti-MM therapies. Cell viability/apoptosis assays were performed using MTT, trypan blue exclusion, and Annexin V/PI staining. Angiogenesis was measured in vitro using Matrigel capillary-like tube structure formation assays: Since human vascular endothelial cells (HUVECs) plated onto Matrigel differentiate and form capillary-like tube structures similar to in vivo neovascularization, this assay measures anti-angiogenic effects of drugs/agents. Migration assays were performed using transwell insert assays. Immunoblot analysis was performed using antibodies to caspase-8, caspase-9, caspase-3, PARP, Bcl-2, Bax, pJNK and GAPDH. Statistical significance was determined using a Student t test. Results Treatment of MM.1S, RPMI-8226, MM.1R, INA-6, and KMS-12BM with NPI-2358 for 24h induces a dose-dependent significant (P < 0.005) decrease in viability of all cell lines (IC50 range: 5-8 nM; n=3). To determine whether NPI-2358-induced decrease in viability is due to apoptosis, MM cell lines were treated with NPI-2358 for 24h; harvested, and analyzed for apoptosis using Annexin V/PI staining. A significant increase in NPI-2358-induced apoptosis was observed in all MM cell lines (% Annexin V+/PI- apoptotic cells: MM.1S, 48 ± 2.3%; MM.1R, 46.6 ± 3.1%; RPMI-8226, 61.7 ± 4.5%; and INA-6, 59.9 ± 3.2%; P < 0.05; n=3). Importantly, NPI-2358 decreased viability of freshly isolated MM cells from patients (IC50 range: 3-7 nM; P < 0.005), without affecting the viability of normal peripheral blood mononuclear cells, suggesting specific anti-MM activity and a favorable therapeutic index for NPI-2358. Examination of in vitro angiogenesis using capillary-like tube structure formation assay showed that even low doses of NPI-2358 (7 nM treatment for 12h; IC50: 20 nM at 24h) significantly decreased tubule formation in HUVECs (70-80% decrease; P < 0.05). Transwell insert assays showed a marked reduction in serum-dependent migration of NPI-2358-treated MM cells (42 ± 2.1% inhibition in NPI-2358-treated vs. control; P < 0.05). NPI-2358 at the concentrations tested (5 nM for 12h) in the migration assays did not affect survival of MM cells (> 95% viable cells). A similar anti-migration activity of NPI-2358 was noted against HUVEC cells (48 ± 1.7% decrease in migration; P < 0.05). Mechanistic studies showed that NPI-2358-induced apoptosis was associated with activation of caspase-8, caspase-9, caspase-3 and PARP. Importantly, treatment of MM.1S cells with NPI-2358 (5 nM) triggered phosphorylation of c-Jun amino-terminal kinase (JNK), a classical stress response protein, without affecting Bcl-2 family members Bax and Bcl-2. Blockade of JNK using dominant negative strategy markedly abrogated NPI-2358-induced apoptosis. Conclusion Our preclinical data provide evidence for remarkable anti-angiogenic and anti-tumor activity of NPI-2358 against MM cells, without significant toxicity in normal cells. Ongoing studies are examining in vivo anti-MM activity of NPI-2358 in animal models. Importantly, a Phase-1 study of NPI-2358 as a single agent in patients with advanced malignancies (lung, prostrate and colon cancer) has already established a favorable pharmacokinetic, pharmacodynamic and safety profile; and, a Phase-2 study of the combination of NPI-2358 and docetaxel in non-small cell lung cancer showed encouraging safety, pharmacokinetic and activity data. These findings, coupled with our preclinical studies, provide the framework for the development of NPI-2358-based novel therapies to improve patient outcome in MM. Disclosures: Chauhan: Nereus Pharmaceuticals, Inc: Consultancy. Lloyd:Nereus Pharmaceuticals, In: Employment. Palladino:Nereus Pharmaceuticals, Inc: Employment. Anderson:Nereus Pharmaceuticals, Inc: Consultancy.

scholarly journals Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60

1999 ◽  
Vol 19 (3-4) ◽  
pp. 139-151 ◽  
Author(s):  
Marina Leite ◽  
Margarida Quinta‐Costa ◽  
Pedro Simas Leite ◽  
José Eduardo Guimarães
Keyword(s):  
Flow Cytometry ◽  
Cell Death ◽  
Fluorescence Microscopy ◽  
Trypan Blue ◽  
Critical Evaluation ◽  
Annexin V ◽  
Tunel Assay ◽  
Apoptotic Cells ◽  
Dna Ladder ◽  
Apoptosis And Necrosis

The reliability of eight distinct methods (Giemsa staining, trypan blue exclusion, acridine orange/ethidium bromide (AO/EB) double staining for fluorescence microscopy and flow cytometry, propidium iodide (PI) staining, annexin V assay, TUNEL assay and DNA ladder) for detection and quantification of cell death (apoptosis and necrosis) was evaluated and compared. Each of these methods detects different morphological or biochemical features of these two processes. The comparative analysis of the 8 techniques revealed that AO/EB (read in fluorescence microscopy) provides a reliable method to measure cells in different compartments (or pathways) of cell death though it is very time consuming. PI staining and TUNEL assay were also sensitive in detecting very early signs of apoptosis, but do not allow precise quantification of apoptotic cells. These three methods were concordant in relation to induction of apoptosis and necrosis in HL60 cells with the various UV irradiation time periods tested. Both AO/EB (read by flow cytometry) and annexin V‐FITC/PI failed to detect the same number of early apoptotic cells as the other three methods. Trypan blue is valueless for this purpose. Giemsa and DNA ladder might be useful as confirmatory tests in some situations.

Study on Apoptosis of Human Promyelocytic Leukemia HL-60 Cells Induced by Fucosterol via Death Receptor Pathway

2013 ◽  
Vol 790 ◽  
pp. 607-610 ◽  
Author(s):  
Yu Bin Ji ◽  
Dong Xue Song ◽  
Chen Feng Ji
Keyword(s):  
Laser Scanning ◽  
Caspase 3 ◽  
Death Receptor ◽  
Promyelocytic Leukemia ◽  
Caspase 8 ◽  
Laser Scanning Confocal Microscope ◽  
Death Receptor Pathway ◽  
Mtt Method ◽  
Induction Of Apoptosis ◽  
Anti Tumor Activity

The purpose of this study is to investigate the effect of fucosterol on the induction of apoptosis and the molecular mechanism involved in Human promyelocytic leukemia HL-60 Cells. HL-60 Cells were treated with different concentrations of fucosterol at different time. MTT method was used to study fucosterol anti-tumor activity. Morphology observation was performed to determine the effects of fucosterol on apoptosis of HL-60 cells. Flow cytometry (FCM) was used to detect the cell cycle. Laser scanning confocal microscope (LSCM) was used to analyze the expressions of Fas, FasL, Fadd and Caspase-8. Caspase activity kits were used to determine the activity of Caspase-8 and Caspase-3. The results showed fucosterol could inhibit the growth of HL-60 cells, and the apoptosis morphology for 48 h treatment was obvious, which showed cell protuberance, cytoplasm concentrated and apoptotic body. Fucosterol treatment for 24 h increased the protein expression of Fas, FasL, Fadd and Caspase-8. It also showed that the activity of Caspase-3 and Caspase-8 has increased significantly. In conclusion, Fucosterol could induce HL-60 cells apoptosis via death receptor pathway.

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