Estimation of cell membrane alteration after drug treatment by LDH release

Jurišić, Vladimir

doi:10.1182/blood-2002-08-2397

Cited by 27 publications

(12 citation statements)

References 12 publications

(25 reference statements)

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“…The overall description of the differentiating treatments was outlined in Figure , together with some characteristics acquired by the cells and relevant for the discussion of the results (i.e., growth rate, cardiac markers, mitochondrial mass, and mitochondrial F 1 F O ATP synthase assembly). During the treatment, cytotoxicity was verified by a sensitive LDH‐release assay, as widely reported for several cell systems [Jurisic, ; Jurisic et al, ]. Very low treatment cytotoxicity was observed (Fig.…”

Section: Resultssupporting

confidence: 57%

F₁F_OATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation

et al. 2015

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Taking advantage from the peculiar features of the embryonic rat heart-derived myoblast cell line H9c2, the present study is the first to provide evidence for the expression of F1FO ATP synthase and of ATPase Inhibitory Factor 1 (IF1) on the surface of cells of cardiac origin, together documenting that they were affected through cardiac-like differentiation. Subunits of both the catalytic F1 sector of the complex (ATP synthase-β) and of the peripheral stalk, responsible for the correct F1-FO assembly/coupling, (OSCP, b, F6) were detected by immunofluorescence, together with IF1. The expression of ATP synthase-β, ATP synthase-b and F6 were similar for parental and differentiated H9c2, while the levels of OSCP increased noticeably in differentiated cells, where the results of in situ Proximity Ligation Assay were consistent with OSCP interaction within ecto-F1FO complexes. An opposite trend was shown by IF1 whose ectopic expression appeared greater in the parental H9c2. Here, evidence for the IF1 interaction with ecto-F1FO complexes was provided. Functional analyses corroborate both sets of data. i) An F1FO ATP synthase contribution to the exATP production by differentiated cells suggests an augmented expression of holo-F1FO ATP synthase on plasma membrane, in line with the increase of OSCP expression and interaction considered as a requirement for favoring the F1-FO coupling. ii) The absence of exATP generation by the enzyme, and the finding that exATP hydrolysis was largely oligomycin-insensitive, are in line in parental cells with the deficit of OSCP and suggest the occurrence of sub-assemblies together evoking more regulation by IF1.

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Section: Resultssupporting

confidence: 57%

F₁F_OATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation

et al. 2015

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“…Among biochemical parameters, LDH has been previously described as an independent prognostic factor, reflecting metabolic disturbance in tumor cells and tumor burden in patients with different malignancies [7,[13][14][15]18]. Further analyses of NK cell activity was based on estimation association of NK cells with elevated serum LDH.…”

Section: Discussionmentioning

confidence: 99%

“…Determination of serum LDH activity LDH activity in MM patients was determined using Ectachem clinical chemistry slides with a multipoint rate slides test (Jonson & Johnson Clinical Diagnositic, USA) on the Ectachem 259 Analyser as previously reported [14,15].…”

Section: Determination Of B 2 -Microglobulinmentioning

confidence: 99%

Clinical stage-depending decrease of NK cell activity in multiple myeloma patients

et al. 2007

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Natural killer cells, as an important subpopulation of cells of the innate immune system have an essential role in defense of the rise and spread of malignancy. These cells have a CD3-CD16 + CD56+ phenotype and they are functionally defined by their ability to lyses tumor cells. We here show that decrease of NK cell activity was significantly associated with advanced clinical stage, increased lactate dehydrogenase (LDH), percentage infiltration of bone marrow with plasma cells, and beta-2 microglobulin. The patients with higher NK cell activity at presentation after receiving VAD protocol have better cumulative survival in comparison with those with low NK cell activity.

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“…This assay is a colorimetric alternative to the 51 Cr release cytotoxicity assay based on the measurement of lactate dehydrogenase (LDH), a stable cytosolic enzyme that is released upon cell lysis. It allows discrimination between effectors and targets LDH release (Konjevic et al, 1997;Jurisic et al, 1999;Jurisic, 2003;Hernandez-Pigeon et al, 2006). Before the assay, NK92 effector cells were cultured in IL-2-free medium containing 12.5% horse serum and 12.5% FBS for 48 h. The target cells used were K562, HepG2, and A549.…”

Section: Cytotoxicity Assaysmentioning

confidence: 99%

Resveratrol enhances perforin expression and NK cell cytotoxicity through NKG2D‐dependent pathways

Chen

2010

Journal Cellular Physiology

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In a previous report, we showed that the in vivo cytotoxic activity of the natural killer (NK) cells isolated from resveratrol-pretreated rats is significantly enhanced compared with that of the non-pretreated rats; however, the underlying mechanism remains unclear. In the present study, we use cultured NK92 cell line to examine the possible signaling pathways underlying the resveratrol-induced activation. Using cultured K562, HepG2, and A549 cells as targets, we show that resveratrol pretreatment increases NK cell cytotoxicity in a dose-dependent manner. The enhanced cytotoxic effect is accompanied by increases in JNK and ERK-1/2 MAP kinase activity and perforin expression. Moreover, the expression of NKG2D, an upstream signaling molecule of the MAP kinases pathway, is also enhanced. Resveratrol-enhanced perforin expression and cytotoxic activity are effectively inhibited by pretreatment with the inhibitors of JNK (SP600125), ERK-1/2 (PD98059), or by siRNAs against JNK-1 and ERK-2. However, the inhibitors or siRNA to p38 exhibits no effect. Since IL-2 has been shown to induce NKG2D expression and perforin release, we therefore, examined whether IL-2 and resveratrol act in parallel. We show that IL-2 also stimulates perforin expression, however, when treated together with resveratrol, they exhibit no additive effect. The results suggest that in NK92 cells, resveratrol may act via a similar or overlapping pathway as that of IL-2, to enhance perforin expression and cytotoxic activity. Data presented strongly indicate that resveratrol act via NKG2D-dependent JNK and ERK-1/2 pathways.

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Estimation of cell membrane alteration after drug treatment by LDH release

Cited by 27 publications

References 12 publications

F₁F_OATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation

F₁F_OATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation

Clinical stage-depending decrease of NK cell activity in multiple myeloma patients

Resveratrol enhances perforin expression and NK cell cytotoxicity through NKG2D‐dependent pathways

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Estimation of cell membrane alteration after drug treatment by LDH release

Cited by 27 publications

References 12 publications

F1FOATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation

F1FOATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation

Clinical stage-depending decrease of NK cell activity in multiple myeloma patients

Resveratrol enhances perforin expression and NK cell cytotoxicity through NKG2D‐dependent pathways

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F₁F_OATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation

F₁F_OATP Synthase Is Expressed at the Surface of Embryonic Rat Heart-Derived H9c2 Cells and Is Affected by Cardiac-Like Differentiation