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Regulation of morphogenesis and neural differentiation of human mesenchymal stem cells using carbon nanotube sheets

In vivo fluorescence imaging of atherosclerotic plaques with activatable cell-penetrating peptides targeting thrombin activity

Diacylglycerol kinase zeta negatively regulates CXCR4-stimulated T lymphocyte firm arrest to ICAM-1 under shear flow

RhoA-induced cytoskeletal tension controls adaptive cellular remodeling to mechanical signaling

Encapsulation of angiogenic monocytes using bio-spraying technology

Synthetic curcumin analog EF31 inhibits the growth of head and neck squamous cell carcinoma xenografts

Systematic analysis of embryonic stem cell differentiation in hydrodynamic environments with controlled embryoid body size

A plant virus substrate induces early upregulation of BMP2 for rapid bone formation

Chromatin accessibility at the HIV LTR promoter sets a threshold for NF-κB mediated viral gene expression

Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining

Preparation, characterization, and surface immobilization of native vesicles obtained by mechanical extrusion of mammalian cells

Resolving brain regions using nanostructure initiator mass spectrometry imaging of phospholipids

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Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining

Content Provider	Royal Society of Chemistry (RSC)
Author	Adam, Vojtech Masarik, Michal Gumulec, Jaromir Kizek, Rene Sztalmachova, Marketa Pavkova-Goldbergova, Monika Hlavna, Marian Cernei, Natalia Sochor, Jiri Zitka, Ondrej Raudenska, Martina Ruttkay-Nedecky, Branislav Krizkova, Sona Babula, Petr
Copyright Year	2012
Abstract	The present paper is focused on zinc(II) treatment effects on prostatic cell lines PC-3 (tumour) and PNT1A (non-tumour). Oxidative status of cells was monitored by evaluation of expression of metallothionein (MT) isoforms 1A and 2A at the mRNA and protein level, glutathione (oxidised and reduced), and intracellular zinc(II) after exposition to zinc(II) treatment at concentrations of 0–150 μM using electrochemical methods, western blotting and fluorescent microscopy. A novel real-time impedance-based growth monitoring system was compared with widely used end-point MTT assay. Impedance-based IC50 for zinc(II) is 55.5 and 150.8 μM for PC-3 and PNT1A, respectively. MTT-determined IC50 are >1.3-fold higher. Impedance-based viability correlates with viable count (r > 0.92; p < 0.03), not with MTT. Two-fold lower intracellular zinc(II) in the tumour PC-3 cell line was found. After zinc(II) treatment >2.6-fold increase of intracellular zinc(II) was observed in non-tumour PNT1A and in tumour PC-3 cells. In PC-3 cells, free and bound zinc(II) levels were enhanced more markedly as compared to PNT1A. PNT1A produced 4.2-fold less MT compared to PC3. PNT1A cells showed a 4.8-fold increase trend (r = 0.94; p = 0.005); PC-3 did show a significant trend at MT1 and MT2 protein levels (r = 0.93; p = 0.02) with nearly ten-fold increase after 100 μM zinc(II) treatment. In terms of redox state, PNT1A had a predominance of reduced GSH forms (GSH : GSSG ratio > 1), when exposed to zinc(II) compared to PC3, where predominance of oxidised forms remains at all concentrations. IC50 differs significantly when determined by MTT and real-time impedance-based assays due to dependence of impedance on cell morphology and adhesion. When real-time growth monitoring, precise electrochemical methods and fluorescent microscopy are performed together, accurate information for metal fluxes, their buffering by thiol compounds and monitoring of the redox state become a powerful tool for understanding the role of oxidative stress in carcinogenesis.
Starting Page	672
Ending Page	684
Page Count	13
File Format	HTM / HTML PDF
ISSN	17579694
Volume Number	4
Issue Number	6
Journal	Integrative Biology
DOI	10.1039/c2ib00157h
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	MTT Zinc PC3 Metallothionein Tonne Messenger RNA Protein Glutathione Electrochemistry Western blot Microscopy MTT assay IC50 Redox Glutathione disulfide Correlation and dependence Electrical impedance Thiol Oxidative stress Carcinogenesis
Content Type	Text
Resource Type	Article
Subject	Medicine Biochemistry Biophysics

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Oxidative metal release from metallothionein via zinc-thiol/disulfide interchange.

Glutamine and kgf each regulate extracellular thiol/disulfide redox and enhance proliferation in caco-2 cells.

Oxidative metal release from metallothionein via zinc-thiol/disulfide interchange.

The effect of tert-butylhydroperoxide on the thiol redox status in human erythrocytes and the protective role of glucose and antioxidants.

Enhanced reactive oxygen species overexpression by CuO nanoparticles in poorly differentiated hepatocellular carcinoma cells

The Glutathione Redox State and Zinc Mobilization from Metallothionein and Other Proteins with Zinc–Sulfur Coordination Sites

Gaussia princeps luciferase: a bioluminescent substrate for oxidative protein folding

Missing links in understanding redox signaling via thiol/disulfide modulation: How is glutathione oxidized in plants?

IC50; MTT assay.

Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining

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