Flow-cytometric analysis of reactive oxygen species in cancer cells under treatment with brassinosteroids

Scope. To elucidate the morphological and biochemicalin vitroeffects exerted by caffeine, taurine, and guarana, alone or in combination, since they are major components in energy drinks (EDs).Methods and Results. On human neuronal SH-SY5Y cells, caffeine (0.125–2 mg/mL), taurine (1–16 mg/mL), and guarana (3.125–50 mg/mL) showed concentration-dependent nonenzymatic antioxidant potential, decreased the basal levels of free radical generation, and reduced both superoxide dismutase (SOD) and catalase (CAT) activities, especially when combined together. However, guarana-treated cells developed signs of neurite degeneration in the form of swellings at various segments in a beaded or pearl chain-like appearance and fragmentation of such neurites at concentrations ranging from 12.5 to 50 mg/mL. Swellings, but not neuritic fragmentation, were detected when cells were treated with 0.5 mg/mL (or higher doses) of caffeine, concentrations that are present in EDs. Cells treated with guarana also showed qualitative signs of apoptosis, including membrane blebbing, cell shrinkage, and cleaved caspase-3 positivity. Flow cytometric analysis confirmed that cells treated with 12.5–50 mg/mL of guarana and its combinations with caffeine and/or taurine underwent apoptosis.Conclusion. Excessive removal of intracellular reactive oxygen species, to nonphysiological levels (or “antioxidative stress”), could be a cause ofin vitrotoxicity induced by these drugs.

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Plasmodium berghei Resists Killing by Reactive Oxygen Species

Infection and Immunity ◽

10.1128/iai.73.10.6704-6710.2005 ◽

2005 ◽

Vol 73 (10) ◽

pp. 6704-6710 ◽

Cited By ~ 27

Author(s):

Peter Sobolewski ◽

Irene Gramaglia ◽

John A. Frangos ◽

Marcos Intaglietta ◽

Henri van der Heyde

Keyword(s):

Reactive Oxygen Species ◽

Plasmodium Berghei ◽

Defense Mechanisms ◽

Flow Cytometric Analysis ◽

Reactive Oxygen ◽

Hemoglobin Concentration ◽

Hemoglobin Level ◽

Disease Development ◽

Oxygen Species ◽

Flow Cytometric

ABSTRACT Reactive oxygen species (ROS) are widely believed to kill malarial parasites. C57BL/6 mice injected with P. berghei inocula incubated with supraphysiological doses of NO (≤150 μM) or with peroxynitrite (220 μM), however, exhibited parasitemia similar to that seen with those given control inocula, and there was no difference in disease development. Only treatment of inocula with NO doses nearing saturation (≥1.2 mM) resulted in no detectable parasitemia in the recipients; flow cytometric analysis with a vital dye (hydroethidine) indicated that 1.5 mM NO lysed the erythrocytes rather than killing the parasites. The hemoglobin level in the inocula was about 8 μM; the hemoglobin was mainly oxyhemoglobin (oxyHb) (96%), which was converted to methemoglobin (>95%) after treatment with 150 μM NO. The concentrations of 150 μM of NO and 220 μM of peroxynitrite were far in excess of the hemoglobin concentration (∼8 μM), and yet no parasite killing was detected. We therefore conclude that hemoglobin protects Plasmodium parasites from ROS, but the parasite likely possesses intrinsic defense mechanisms against ROS.

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Reserpine Induces Apoptosis and Cell Cycle Arrest in Hormone Independent Prostate Cancer Cells through Mitochondrial Membrane Potential Failure

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520618666180209152215 ◽

2019 ◽

Vol 18 (9) ◽

pp. 1313-1322 ◽

Cited By ~ 5

Author(s):

Manjula Devi Ramamoorthy ◽

Ashok Kumar ◽

Mahesh Ayyavu ◽

Kannan Narayanan Dhiraviam

Keyword(s):

Prostate Cancer ◽

Reactive Oxygen Species ◽

Cell Cycle ◽

Membrane Potential ◽

Cancer Cells ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Reactive Oxygen ◽

Oxygen Species ◽

Prostate Cancer Cells

Background: Reserpine, an indole alkaloid commonly used for hypertension, is found in the roots of Rauwolfia serpentina. Although the root extract has been used for the treatment of cancer, the molecular mechanism of its anti-cancer activity on hormonal independent prostate cancer remains elusive. Methods: we evaluated the cytotoxicity of reserpine and other indole alkaloids, yohimbine and ajmaline on Prostate Cancer cells (PC3) using MTT assay. We investigated the mechanism of apoptosis using a combination of techniques including acridine orange/ethidium bromide staining, high content imaging of Annexin V-FITC staining, flow cytometric quantification of the mitochondrial membrane potential and Reactive Oxygen Species (ROS) and cell cycle analysis. Results: Our results indicate that reserpine inhibits DNA synthesis by arresting the cells at the G2 phase and showed all standard sequential features of apoptosis including, destabilization of mitochondrial membrane potential, reduced production of reactive oxygen species and DNA ladder formation. Our in silico analysis further confirmed that indeed reserpine docks to the catalytic cleft of anti-apoptotic proteins substantiating our results. Conclusion: Collectively, our findings suggest that reserpine can be a novel therapeutic agent for the treatment of androgen-independent prostate cancer.

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ent-Kaurane Diterpenoids from Croton tonkinensis Induce Apoptosis in Colorectal Cancer Cells through the Phosphorylation of JNK Mediated by Reactive Oxygen Species and Dual-Specificity JNK Kinase MKK4

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520614666140127111407 ◽

2014 ◽

Vol 14 (7) ◽

pp. 1051-1061 ◽

Cited By ~ 8

Author(s):

Phuong Thuong ◽

Nguyen Khoi ◽

Saho Ohta ◽

Shinichiro Shiota ◽

Hironori Kanta ◽

...

Keyword(s):

Colorectal Cancer ◽

Reactive Oxygen Species ◽

Cancer Cells ◽

Reactive Oxygen ◽

Oxygen Species ◽

Dual Specificity ◽

Colorectal Cancer Cells

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Flow Cytometric Detection of Reactive Oxygen Species

BIO-PROTOCOL ◽

10.21769/bioprotoc.431 ◽

2013 ◽

Vol 3 (8) ◽

Cited By ~ 2

Author(s):

Hsin-Yi Chang ◽

Hsuan-Cheng Huang ◽

Tsui-Chin Huang ◽

Pan-Chyr Yang ◽

Yi-Ching Wang ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Reactive Oxygen ◽

Oxygen Species ◽

Flow Cytometric

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Photo-Responsive Supramolecular Micelles for Controlled Drug Release and Improved Chemotherapy

International Journal of Molecular Sciences ◽

10.3390/ijms22010154 ◽

2020 ◽

Vol 22 (1) ◽

pp. 154

Author(s):

Fasih Bintang Ilhami ◽

Kai-Chen Peng ◽

Yi-Shiuan Chang ◽

Yihalem Abebe Alemayehu ◽

Hsieh-Chih Tsai ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Drug Release ◽

Visible Light ◽

Cancer Cells ◽

Laser Irradiation ◽

Reactive Oxygen ◽

Controlled Drug Release ◽

Grand Challenge ◽

Supramolecular System ◽

Oxygen Species

Development of stimuli-responsive supramolecular micelles that enable high levels of well-controlled drug release in cancer cells remains a grand challenge. Here, we encapsulated the antitumor drug doxorubicin (DOX) and pro-photosensitizer 5-aminolevulinic acid (5-ALA) within adenine-functionalized supramolecular micelles (A-PPG), in order to achieve effective drug delivery combined with photo-chemotherapy. The resulting DOX/5-ALA-loaded micelles exhibited excellent light and pH-responsive behavior in aqueous solution and high drug-entrapment stability in serum-rich media. A short duration (1–2 min) of laser irradiation with visible light induced the dissociation of the DOX/5-ALA complexes within the micelles, which disrupted micellular stability and resulted in rapid, immediate release of the physically entrapped drug from the micelles. In addition, in vitro assays of cellular reactive oxygen species generation and cellular internalization confirmed the drug-loaded micelles exhibited significantly enhanced cellular uptake after visible light irradiation, and that the light-triggered disassembly of micellar structures rapidly increased the production of reactive oxygen species within the cells. Importantly, flow cytometric analysis demonstrated that laser irradiation of cancer cells incubated with DOX/5-ALA-loaded A-PPG micelles effectively induced apoptotic cell death via endocytosis. Thus, this newly developed supramolecular system may offer a potential route towards improving the efficacy of synergistic chemotherapeutic approaches for cancer.

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