Cytometric analysis for drug-induced steatosis in HepG2 cells

Donato, M. Teresa; Martínez-Romero, Alicia; Jiménez, Núria; Negro, Alejandro; Herrera, Guadalupe; Castell, José V.; O’Connor, José-Enrique; Gómez‐Lechón, M. José

doi:10.1016/j.cbi.2009.07.019

Cited by 80 publications

(81 citation statements)

References 35 publications

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“…A similar decrease in mitochondrial capacity was observed in endothelial cells (Dranka et al 2010). This profile indicates that DMNQ can generate an enormous flux of hydrogen peroxide, and generation of high levels of reactive oxygen species (ROS) is characteristic of redox agents compared with other toxicants (Donato et al 2009;Schoonen et al 2005;Xu et al 2008).…”

Section: Dimethoxynaphthoquinone (Dmnq)supporting

confidence: 56%

SEURAT-1 liver gold reference compounds: a mechanism-based review

et al. 2014

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There is an urgent need for the development of alternative methods to replace animal testing for the prediction of repeat dose chemical toxicity. To address this need, the European Commission and Cosmetics Europe have jointly funded a research program for 'Safety Evaluation Ultimately Replacing Animal Testing.' The goal of this program was the development of in vitro cellular systems and associated computational capabilities for the prediction of hepatic, cardiac, renal, neuronal, muscle, and skin toxicities. An essential component of this effort is the choice of appropriate reference compounds that can be used in the development and validation of assays. In this review, we focus on the selection of reference compounds for liver pathologies in the broad categories of cytotoxicity and lipid disorders. Mitochondrial impairment, oxidative stress, and apoptosis are considered under the category of cytotoxicity, while steatosis, cholestasis, and phospholipidosis are considered under the category of lipid dysregulation. We focused on four compound classes capable of initiating such events, i.e., chemically reactive compounds, compounds with specific cellular targets, compounds that modulate lipid regulatory networks, and compounds that disrupt the plasma membrane. We describe the molecular mechanisms of these compounds and the cellular response networks which they elicit. This information will be helpful to both improve our understanding of mode of action and help in the selection of appropriate mechanistic biomarkers, allowing us to progress the development of animal-free models with improved predictivity to the human situation.

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Section: Dimethoxynaphthoquinone (Dmnq)supporting

confidence: 56%

SEURAT-1 liver gold reference compounds: a mechanism-based review

et al. 2014

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“…Cells were stained with 2 0 ,7 0 -dihydrochlorofluorescein diacetate (DHCF-DA, Sigma-Aldrich) as in ref. 20. All samples were analyzed on a CyAnADP Flow Cytometer, using the Summit 4.3 software (Beckman Coulter).…”

Section: Facs Analysismentioning

confidence: 99%

Immunotargeting of Antigen xCT Attenuates Stem-like Cell Behavior and Metastatic Progression in Breast Cancer

Lanzardo

Conti

Rooke³

et al. 2016

Cancer Research

138

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Resistance to therapy and lack of curative treatments for metastatic breast cancer suggest that current therapies may be missing the subpopulation of chemoresistant and radioresistant cancer stem cells (CSC). The ultimate success of any treatment may well rest on CSC eradication, but specific anti-CSC therapies are still limited. A comparison of the transcriptional profiles of murine Her2 þ breast tumor TUBO cells and their derived CSC-enriched tumorspheres has identified xCT, the functional subunit of the cystine/glutamate antiporter system x c À , as a surface protein that is upregulated specifically in tumorspheres. We validated this finding by cytofluorimetric analysis and immunofluorescence in TUBO-derived tumorspheres and in a panel of mouse and human triple negative breast cancer cell-derived tumorspheres. We further show that downregulation of xCT impaired tumorsphere generation and altered CSC intracellular redox balance in vitro, suggesting that xCT plays a functional role in CSC biology. DNA vaccination based immunotargeting of xCT in mice challenged with syngeneic tumorsphere-derived cells delayed established subcutaneous tumor growth and strongly impaired pulmonary metastasis formation by generating anti-xCT antibodies able to alter CSC self-renewal and redox balance. Finally, anti-xCT vaccination increased CSC chemosensitivity to doxorubicin in vivo, indicating that xCT immunotargeting may be an effective adjuvant to chemotherapy. Cancer Res; 76(1); 62-72. Ó2015 AACR.

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“…Considering the absence of changes in HDL-c in the short term, we monitored the animals for 13 weeks and found that the treated groups showed a significant increase in TC, LDL-c, and HDL-c. By promoting cholesterol accumulation and inhibiting lipid metabolism, AM can decrease the abundance of LDL receptors in hepatic cells at both mRNA and protein levels [12][13][14]. The decreased LDL receptor levels disturb the respiratory electron transport chain, prevent the transformation of TG to VLDL, and inhibit both PPARα activity and β-oxidization [15]; this results in changes in the mitochondrial membrane potential due to the accumulation of TG and lipid droplets in hepatic cells, and this accumulation may lead to non-alcoholic fatty liver disease [12][13][14][15][16][17][18][19][20][21].…”

Section: Discussionmentioning

confidence: 99%

Dronedarone and Amiodarone Induce Dyslipidemia and Thyroid Dysfunction in Rats

Jiang

Chen

et al. 2016

Cell Physiol Biochem

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Background/Aims: Amiodarone, a thyroid hormone-like molecule, can induce dyslipidemia and thyroid dysfunction. However, the effects of dronedarone on lipid metabolism and of both dronedarone and amiodarone on thyroid function and lipid metabolism remain unknown. Methods: Fifty male Sprague-Dawley rats were randomly divided into 5 groups (10 in each group): normal control (NC), amiodarone-treated (AMT), dronedarone-treated (DRT), rats treated with amiodarone combined with polyene phosphatidylcholine (AC), and rats treated with dronedarone combined with polyene phosphatidylcholine (DC). Rats were given amiodarone (120 mg/kg/d), dronedarone (120 mg/kg/d), and polyene phosphatidylcholine (200 mg/kg/d) for 13 weeks. At the end of weeks 4, 8, 12, and 13, plasma-free triiodothyronine (FT3), free thyroxine (FT4), triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), and high-density lipoprotein cholesterol (HDL-c) were determined. At the end of this protocol, rats were sacrificed and the thyroid glands were isolated, weighed, and examined histopathologically. The protein expression of Bcl-2 was measured by immunochemical staining. The mRNA expression of thyroglobulin (Tg), type-1 deiodinase (D1), and thyroid peroxidase (TPO) were detected by polymerase chain reaction (PCR). Results: Compared with the NC group, FT3 and FT4 levels in the DRT and DC groups significantly increased at week 4 but declined thereafter. The AMT and AC groups had lower FT3 levels but comparable FT4 levels. The levels of TG, LDL-c, and HDL-c in the NC group were lower than those in the other groups whereas the LDL-c/HDL-c ratio was lowest in the AMT group. Bcl-2 expression significantly increased in the DRT group. The mRNA expression of Tg increased whereas the mRNA expression of D1 decreased. Dronedarone induced hyperthyroidism at the early stage and hypothyroidism at the late stage whereas amiodarone only caused hypothyroidism. Conclusion: Both dronedarone and amiodarone can induce dyslipidemia and increase the levels of TC, LDL-c, and HDL-c, and these effects may be associated with thyroid dysfunction.

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Cytometric analysis for drug-induced steatosis in HepG2 cells

Cited by 80 publications

References 35 publications

SEURAT-1 liver gold reference compounds: a mechanism-based review

SEURAT-1 liver gold reference compounds: a mechanism-based review

Immunotargeting of Antigen xCT Attenuates Stem-like Cell Behavior and Metastatic Progression in Breast Cancer

Dronedarone and Amiodarone Induce Dyslipidemia and Thyroid Dysfunction in Rats

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