In Vitro Evaluation of Hepg2 Cell Proliferation Altered by Reactive Oxygen and Nitrogen Species

Pengcheng Li

doi:10.21767/2572-309x.1000112

Abstract

This study was to investigate cell proliferation regulated by reactive oxygen and nitrogen species and their scavengers. Earlier conclusions are paradoxical on roles that reactive oxygen and nitrogen species and their scavengers enhance or inhibit cancer cell proliferations. This study employed the MTS assay to evaluate proliferations of HepG2 cells treated with various concentrations of two different nitric oxide donors, hydrogen peroxide and their scavengers for 24 hours. The MTS assay revealed that low concentrations of SNP, SNAP and H2O2 can significantly enhance HepG2 cell proliferation, and high concentrations of cPTIO significantly inhibited HepG2 cell proliferation. Additionally, the assay also indicated that 100 μM H2O 2 , 100 μM cPTIO or 100 U/mL catalase did not have evident synergisms with NSP or SNAP. However, due to reactions of the chemicals with reagents of kits of the MTS assay, the data were not inferred that low concentrations of NO and H2O2 enhance proliferation, whereas high concentrations inhibit proliferation, and whether there are synergistic effects with the combination of SNP (or SNAP) and H2O2 , cPTIO or catalase.

Highlights

The hallmark of hepatocellular carcinoma (HCC) is uncontrolled proliferation of tumour cells, which ignore inhibitory signals to cell proliferation, promote growth of blood vessels and invade other organs [1]
Statistical differences of proliferation rates of HepG2 cells treated with various concentrations of SNP, SNAP, H2O2 or cPTIO compared to those of negative controls that were not treated with these chemicals were determined using one-way analysis of variance followed by a Dunnett's Multiple Comparison Test
There was a significant increase in cell proliferation at doses of SNAP between 1 to 100 μM SNAP, but there was no significant difference from 200 to 1000 μM SNAP compared to negative controls (Figure 1B)

Summary

Introduction

The hallmark of hepatocellular carcinoma (HCC) is uncontrolled proliferation of tumour cells, which ignore inhibitory signals to cell proliferation, promote growth of blood vessels and invade other organs [1]. Mounting evidence suggests that reactive oxygen species (ROS) and reactive nitrogen species (RNS) generated through chronic liver inflammation are involved in the process of HCC development [4,5,6]. Epidemiology has revealed that most of HCCs develop in chronic inflammatory conditions of the liver [1]. In these conditions, various activated inflammatory and immune cells are recruited to the site of inflammation and generate ROS and RNS. Local concentrations of ROS and RNS have been shown to be elevated in chronic liver inflammation [7,8,9,10]

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Conclusion