The Evidence of HeLa Cell Apoptosis Induced with Tetraethylammonium Using Proteomics and Various Analytical Methods

Huang, Lin; Huang, Qingyu; Huang, Heqing

doi:10.1074/jbc.m113.515932

Cited by 12 publications

(9 citation statements)

References 63 publications

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“…The osmolarity of 30 mM TEA in TCM is ~16% higher than control TCM; however, the toxic effects of 30 mM TEA is not due to high osmolarity as the migration and survival of ENCCs in intact explants of E12.5 gut grown in TCM with 19% higher osmolality due to added sucrose did not differ from explants grown in control TCM (data not shown). Our data are consistent with very recent studies showing that exposure to high concentrations of TEA (>2 mM) kills HeLa cells [ 56 ] and cultured hippocampal neurons [ 57 ]. Lower concentrations of TEA (2 mM, 10 mM) or 4-AP (0.1 mM) had no significant effect on ENCC migration ( Fig 4C ), neuritogenesis ( Fig 4D ) or cell death ( Fig 4F ).…”

Section: Resultssupporting

confidence: 93%

“…It is therefore highly likely that the decreased neuritogenesis observed after 9 h exposure to 30 mM TEA or 5 mM 4-AP is due to ENCCs being unhealthy. Other recent studies using HeLa cells and cultured hippocampal neurons have also reported that exposure to high concentrations of TEA for 24 h or more results in cell death [ 56 , 57 ]. Proteomics analyses of HeLa cells exposed to high concentrations of TEA revealed changes in proteins involved in a variety of biological functions including oxidative stress responses, protein synthesis and degradation, metabolism and signal transduction [ 56 ].…”

Section: Discussionmentioning

confidence: 99%

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Ion Channel Expression in the Developing Enteric Nervous System

et al. 2015

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The enteric nervous system arises from neural crest-derived cells (ENCCs) that migrate caudally along the embryonic gut. The expression of ion channels by ENCCs in embryonic mice was investigated using a PCR-based array, RT-PCR and immunohistochemistry. Many ion channels, including chloride, calcium, potassium and sodium channels were already expressed by ENCCs at E11.5. There was an increase in the expression of numerous ion channel genes between E11.5 and E14.5, which coincides with ENCC migration and the first extension of neurites by enteric neurons. Previous studies have shown that a variety of ion channels regulates neurite extension and migration of many cell types. Pharmacological inhibition of a range of chloride or calcium channels had no effect on ENCC migration in cultured explants or neuritogenesis in vitro. The non-selective potassium channel inhibitors, TEA and 4-AP, retarded ENCC migration and neuritogenesis, but only at concentrations that also resulted in cell death. In summary, a large range of ion channels is expressed while ENCCs are colonizing the gut, but we found no evidence that ENCC migration or neuritogenesis requires chloride, calcium or potassium channel activity. Many of the ion channels are likely to be involved in the development of electrical excitability of enteric neurons.

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Section: Resultssupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Ion Channel Expression in the Developing Enteric Nervous System

et al. 2015

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“…We did not observe any significant changes in cell viability after 24h or 48 h incubation with apamin or TRAM-34. At the same time, treatment of the cells with 10 mM TEA slightly lowered the viability of K562 cells after 48 h. The more significant effect of 10 mM TEA on cancer cell viability was reported earlier [46]: about 50% of cervical cancer HeLa cells were dead after treatment with 10 mM TEA for 48 h. In our assay, 10 mM TEA almost completely blocked cell proliferation after 24 h and 48 h of incubation, whereas negative effect on cell viability was observed only after 48 h. The inhibition of proliferation by TEA was previously shown on C6, 9L glioma, cervical carcinoma SiHA and endometrial adenocarcinoma HEC1-A cell lines [47]. In glioma cells, treatment of the cells with TEA significantly increased the number of reactive oxygen species (ROS) and alteration of B-cell lymphoma protein 2/B-cell lymphoma protein-associated X (Bcl-2/Bax) balance, which further resulted in cell apoptosis [48].…”

Section: Discussionsupporting

confidence: 73%

Role of Calcium-Activated Potassium Channels in Proliferation, Migration and Invasion of Human Myeloid Leukemia Cells

Vasileva¹,

Sudarikova²,

Khairullina³

et al. 2023

Preprint

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Calcium-activated potassium channels (KCa) are the important participants in calcium signaling pathways due to their ability to be activated by increase of the intracellular free calcium concentration. KCa channels are involved in the regulation of various processes in the cells under normal, as well as pathophysiological conditions, including oncotransformation. Previously, with the use of patch-clamp method, we registered the KCa activity in the plasma membrane of human myeloid leukemia K562 cell line. Here, we performed the molecular and functional identification of KCa channels and have uncovered their role in proliferation, migration and invasion of K562 cells. Using a combined approach, we identified the functional activity of SK2, SK3 and IK channels in the plasma membrane of the cells. Selective SK and IK channel inhibitors, apamin and TRAM-34, reduced the proliferative, migratory and invasive capabilities of human myeloid leukemia cells. At the same time, the viability of K562 cells was not affected by KCa channel inhibitors. Our data imply that SK/IK channel inhibitors could be used to slow down the proliferation and spreading of leukemia cells that express functionally active KCa channels in the plasma membrane.

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“…Additionally, the addition of 2.0 mM glycyrrhizin did not have an additive effect with cisplatin in suppressing NCI-H23 cell growth. Cisplatin can induce apoptosis at 2 μg/ml in several cancer cell lines [24,28], and the IC50 of cisplatin for A549 is approximately 12 μM, i.e., 3.6 μg/ml [29]. Therefore, 2.5 μg/ml and 4 μg/ml of cisplatin were used in this study.…”

Section: Resultsmentioning

confidence: 99%

Glycyrrhizin Suppresses Lung Adenocarcinoma Cell Growth Through Inhibition of Thromboxane Synthase

Huang¹,

Chu²,

Jiang

et al. 2014

Cell Physiol Biochem

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Background/Aims: The effects of glycyrrhizin treatment in lung cancer remain undetermined, despite extensive studies of the anti-tumor activities of glycyrrhizin. Methods: Lung adenocarcinoma A549 and NCI-H23 cell lines were used in this study. Cell growth was examined by MTS assays, while apoptosis and cell cycle were determined by flow cytometric analysis. Both real-time PCR and western blotting were used to examine the expression levels of thromboxane synthase (TxAS), and TxAS activity was measured using EIA detection of the biosynthesis of TxA2. TxAS was overexpressed in NCI-H23 cells by transfection with TxAS cDNA, while TxAS was inhibited by transfection with TxAS siRNA in A549 cells. For the mouse model of lung adenocarcinoma, the effects of glycyrrhizin on tumor growth were analyzed by western blot evaluation of TxAS, PTEN and survivin. TxAS activity was determined by EIA assay. Results: Glycyrrhizin suppressed cell growth in A549 cells, but not in NCI-H23 cells, by induction of apoptosis. TxAS was overexpressed in A549 cells, but the TxAS levels in NCI-H23 cells were minimal. Moreover, TxAS expression and activity were suppressed by glycyrrhizin. Glycyrrhizin had no additive effects with TxAS siRNA knockdown in suppressing A549 cell growth, whereas it completely suppressed cell growth of NCI-H23 cells transfected with TxAS cDNA. These results were further confirmed by the in vivo study. Conclusion: Our study suggests that the anti-tumor effect of glycyrrhizin in lung adenocarcinoma is, at least in part, TxAS-dependent. Therefore, glycyrrhizin is a promising anti-cancer agent for the treatment of lung adenocarcinoma.

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The Evidence of HeLa Cell Apoptosis Induced with Tetraethylammonium Using Proteomics and Various Analytical Methods

Cited by 12 publications

References 63 publications

Ion Channel Expression in the Developing Enteric Nervous System

Ion Channel Expression in the Developing Enteric Nervous System

Role of Calcium-Activated Potassium Channels in Proliferation, Migration and Invasion of Human Myeloid Leukemia Cells

Glycyrrhizin Suppresses Lung Adenocarcinoma Cell Growth Through Inhibition of Thromboxane Synthase

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