Role of calcium ion channels and cytoskeletal proteins in Thorium-232 induced toxicity in normal human liver cells (WRL 68) and its validation in swiss mice

被引：0

作者：

Yadav, Rakhee ^{[1
,2
]}

Das, Sourav Kumar ^{[1
]}

Ali, Manjoor ^{[1
]}

Pandey, Badri N. ^{[1
,2
]}

Kumar, Amit ^{[1
,2
]}

机构：

[1] Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Trombay, Mumbai,400 085, India

[2] Homi Bhabha National Institute, Anushaktinagar, Mumbai,400 094, India

来源：

Chemosphere | 2022年 / 288卷

关键词：

Cell proliferation - Chelation - Calcium - Proteins - Cell membranes - High resolution transmission electron microscopy - Inductively coupled plasma mass spectrometry - Ions - Mammals - Thorium;

D O I：

暂无

中图分类号：

学科分类号：

摘要：

Hepatic disorders reported in humans exposed to Thorium-232 (Th-232) rationalizes the present study investigating the toxicological response of normal human liver cells (WRL 68) and its validation in Swiss mice. Cell count analysis of WRL 68 cells-treated with Th-nitrate (1–200 μM) estimated IC50 of ∼24 μM (at 24 h) and 35 μM (at 48 h). Analysis of cell viability (trypan blue assay) showed the IC50 of ∼172 μM. Phase contrast bright-field microscopy revealed Th-induced morphological changes and cell-released microvesicle-like structures in extracellular space. Th-estimation by ICP-MS (Inductively-coupled plasma mass-spectrometry) showed uptake of Th by cells as a function of concentration and incubation time. Employing DTPA as a chelating agent in cell harvesting solution, cell-internalized/strongly-bound Th was estimated to be ∼42% of total incubated Th. Th-uptake studies in the presence of ion-channel specific inhibitors (e.g. nifedipine, thapsigargin) revealed the role of plasma membrane calcium channels and cytoplasmic calcium in modulating the Th-uptake. Transmission electron microscopy of Th-treated cells showed cell-derived extracellular vesicles, alterations in the shape and size of nucleus and mitochondria as well as cytoplasmic inclusions. The order of Th accumulation in various sub-cellular protein fractions was found to be as cytoskeleton (43%) > cytoplasmic (15%) > chromatin (7%) > nuclear (5%) & membrane (5%). Immunofluorescence analysis of WRL 68 cells showed that Th significantly altered the expression of cytoskeleton proteins (F-actin and keratin), which was further validated in liver tissues of Swiss mice administered with Th-232. Findings herein highlight the role of calcium channels and cytoskeleton in Th-induced toxicity. Keywords: Thorium toxicity; Liver cells; Calcium channels; Sub-cellular targets, Cytoskeleton; Swiss Mice. © 2021 Elsevier Ltd

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