Pyroptotic and apoptotic cell death in iNOS and nNOS overexpressing K562 cells: A mechanistic insight

Sadaf, Samreen; Awasthi, Deepika; Singh, Abhishek Kumar; Nagarkoti, Sheela; Kumar, Sachin; Barthwal, Manoj Kumar; Dikshit, Madhu

doi:10.1016/j.bcp.2019.113779

Cited by 10 publications

(8 citation statements)

References 91 publications

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“…Apart from ROS, the mitochondria are a major source of RNS in living cells [ 46 ]. Research has revealed that RNS promote pyroptosis [ 47 ]. It is also probable that autophagy inhibits pyroptosis through clearance of mitochondrial RNS.…”

Section: Autophagy Is a Negative Modulator Of Pyroptosismentioning

confidence: 99%

Autophagy Regulation on Pyroptosis: Mechanism and Medical Implication in Sepsis

Guo

Wang

Cui

2021

Mediators of Inflammation

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Sepsis is defined as a life-threatening disease involving multiple organ dysfunction caused by dysregulated host responses to infection. To date, sepsis remains a dominant cause of death among critically ill patients. Pyroptosis is a unique form of programmed cell death mediated by the gasdermin family of proteins and causes lytic cell death and release of proinflammatory cytokines. Although there might be some positive aspects to pyroptosis, it is regarded as harmful during sepsis and needs to be restricted. Autophagy was originally characterized as a homeostasis-maintaining mechanism in living cells. In the past decade, its function in negatively modulating pyroptosis and inflammation during sepsis has attracted increased attention. Here, we present a comprehensive review of the regulatory effect of autophagy on pyroptosis during sepsis, including the latest advances in our understanding of the mechanism and signaling pathways involved, as well as the potential therapeutic application in sepsis.

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Section: Autophagy Is a Negative Modulator Of Pyroptosismentioning

confidence: 99%

Autophagy Regulation on Pyroptosis: Mechanism and Medical Implication in Sepsis

Guo

Wang

Cui

2021

Mediators of Inflammation

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“…Our group has shown that the overexpression of NOS isoform induced cytostasis and apoptosis in human leukemic K562 cells 28 ; therefore, the role of iNOS on imatinib response was further validated using iNOS stably transfected K562 cells (K562 iNOS ). Stable overexpression of iNOS was confirmed by enhanced iNOS mRNA and protein expression leading to reduced cell cycle progression, wheras expression of BCR–ABL was impervious in these cells (Figs.…”

Section: Resultsmentioning

confidence: 99%

“…Analysis of K562 cells apoptosis was assessed using Annexin V‐FITC/Propidium iodide Kit (BD Biosciences, San Jose, CA, USA). Stained cells were acquired by flow cytometer (FACS Calibur; Becton Dickinson, Franklin Lakes, NJ, USA) and analyzed by using Cell Quest software (Becton Dickinson, Franklin Lakes, NJ, USA) 28 …”

Section: Methodsmentioning

confidence: 99%

“…Mitochondrial membrane potential was measured using JC‐1 (5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetra‐ethyl‐benzimidazocarbo‐cyaniniodide), a cationic dye, which accumulate in mitochondria in a membrane potential‐dependent manner. Fluorescence of 10,000 cells was acquired by FACS Calibur (Becton Dickinson, Franklin Lakes, NJ, USA) and subsequently analyzed using Cell Quest programme (Becton Dickinson, Franklin Lakes, NJ, USA) 28 …”

Section: Methodsmentioning

confidence: 99%

“…Protein (100 μg) was mixed with caspase‐3 substrate acetyl‐Asp‐Glu‐Val‐Asp‐7‐amino‐4 methylcoumarin (150 μM) to assess changes in the caspase‐3 enzyme activity. Fluorescence of cleaved 7‐amino‐4 methylcoumarin (Sigma–Aldrich) was measured using 380 nm (excitation) and 460 nm (emission) wavelengths on a fluorimeter (Omega; BMG Labtech, Ortenberg, Germany) 7,28 …”

Section: Methodsmentioning

confidence: 99%

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Expression of inducible NOS is indispensable for the antiproliferative and proapoptotic effect of imatinib in BCR–ABL positive cells

Singh

Awasthi

Dubey

et al. 2021

Journal of Leukocyte Biology

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Chronic myeloid leukemia (CML) is characterized by constitutive BCR–ABL kinase activity, an aggressive proliferation of immature cells, and reduced differentiation. Targeting tyrosine kinase activity of BCR–ABL with imatinib is an effective therapy for the newly diagnosed CML patients; however, 20%–30% of the patients initially treated with imatinib eventually experience treatment failure. Therefore, early identification of these patients is of high clinical relevance. In the present study, we by undertaking a direct comparison of inducible NOS (iNOS) status in neutrophils from healthy volunteers, newly diagnosed, imatinib responder, and resistant CML patients as well as by conducting in vitro studies in K562 cells demonstrated that inhibition of BCR–ABL by imatinib or siRNA significantly enhanced NO generation and iNOS expression. Indeed, patients exhibiting treatment failure or imatinib resistance were less likely to induce NO generation/iNOS expression. Our findings further demonstrated that imatinib mediated antiproliferative and proapoptotic effect in BCR–ABL+ cells associated with enhanced iNOS expression, and it was significantly prevented in the presence of L‐NAME, 1400W, or iNOS siRNA. Overexpression of iNOS in K562 cells expectedly enhanced imatinib sensitivity on cytostasis and apoptosis, even at lower concentration (0.1 μM) of imatinib. Mechanistically, imatinib or BCR–ABL siRNA following deglutathionylation of NF‐κB, enhanced its binding to iNOS promoter and induced iNOS transcription. Deglutathionylation of procaspase‐3 however associated with increased caspase‐3 activity and cell apoptosis. Taken together, results obtained suggest that monitoring NO/iNOS level could be useful to identify patients likely to be responsive or resistant to imatinib and can be used to personalized alternative therapy.

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