Copper excess in liver HepG2 cells interferes with apoptosis and lipid metabolic signaling at the protein level

Abstract

Background/Aims: Copper is an essential trace element that serves as an important catalytic cofactor for cuproenzymes, carrying out major biological functions in growth and development. Although Wilson's disease (WD) is unquestionably caused by mutations in the ATP7B gene and subsequent copper overload, the precise role of copper in inducing pathological changes remains poorly understood. Materials and Methods: Our study aimed to explore, in HepG2 cells exposed to copper, the cell viability and apoptotic cells was tested by MTT and Hoechst 33342 stainning respectively, and the signaling pathways involved in oxidative stress response, apoptosis and lipid metabolism were determined by real time RT-PCR and Western blot analysis.Results: The results demonstrate dose-and time-dependent cell viability and apoptosis in HepG2 cells following treatment with 10μM , 200μM and 500μM of copper sulfate for 8 and 24 h. Copper overload significantly induced the expression of HSPA1A (heat shock 70kDa protein 1A), an oxidative stress-responsive signal gene, and BAG3 (BCL2 associated athanogene3), an anti-apoptotic gene, while expression of HMGCR (3-hydroxy-3-methylglutarylCoA reductase), a lipid biosynthesis and lipid metabolism gene, was inhibited. Conclusion: These findings provide new insights into possible mechanisms accounting for the development of liver apoptosis and steatosis in the early stages of Wilson's disease.

High copper concentrations produce genotoxicity and cytotoxicity in bovine cumulus cells

“…Also, the accumulation of Cu oxide nanoparticles (CuO-NPs) in C6 glioma cells produces a severe loss in cell viability (assessed by MTT reduction and cellular lactate dehydrogenase activity) and cell membrane integrity (Joshi et al 2016). Similar results have been observed when Cu chloride was used (Nzengue et al 2012); also, HepG2 cell exposure to 10, 200, and 500 μM Cu sulfate for 8 or 24 h reduced the viability as compared with untreated cells (Liu et al 2014). In the present study, loss in cellular MTT reduction capacity was observed for all Cu conditions, suggesting that the MTT assay is a more sensitive indicator of Cu-induced cellular viability decrease than the TB-FDA assay.…”

Antimicrobial Effect of Biocompatible Silicon Nanoparticles Activated Using Therapeutic Ultrasound

“…A typical disadvantage of using nanoparticles is their rather high cytotoxicity. In particular, it was found that metal and metal-oxide nanoparticles, which had been implanted into mammalian cells, caused alteration of the normal function of mitochondria, an increase in the membrane permeability, and the generation of reactive oxygen species, which resulted in oxidative stress and cellular damage. ,− However, nanoparticles that are biocompatible and biodegradable (which strongly depends on their physical–chemical properties) can potentially be used for in vivo applications …”