Screening and validation of nickel ion cytotoxicity biomarkers based on transcriptomic and proteomic technology

Abstract

The aim of this study was to screen cytotoxicity biomarkers of nickel ions (Ni2+) using transcriptomic and proteomic approaches combined with molecular biology validation. First, the MTT method was used to evaluate cytotoxicity in L929 cells treated with Ni2+ at different concentrations. Ni2+ at both 100 μM and 200 μM affected cell proliferation. Then, transcriptomic and proteomic technology was used to study the effects of Ni2+ on the expression of genes/proteins in cells. It was found that 1490, 789, 652 and 729 genes (12, 24, 48 and 72 h, respectively) and 177, 2191 and 2095 proteins (12, 24 and 48 h, respectively) were differentially expressed after treatment with 100 μM Ni2+. In total, 1403, 963, 916 and 1230 genes (12, 24, 48 and 72 h, respectively) and 83, 1681 and 2398 proteins (12, 24 and 48 h, respectively) were differentially expressed after treatment with 200 μM Ni2+. Then, 4 target gene/protein biomarkers were filtered by combined screening using gene/proteomic experimental data and biological pathway analyses. Further expression level validation of all these target biomarkers and functional validation of selected gene/protein biomarkers were carried out, and a final gene/protein biomarker (UQCRB) was identified.

A strategy for mechanically integrating robust hydrogel-tissue hybrid to promote the anti-calcification and endothelialization of bioprosthetic heart valve

“…The cytotoxicity of modified BHVs was evaluated by incubating L929 cells with extracts from different samples for 3 days [ 43 , 44 ]. As shown in Figure 4A , more live cells and barely any dead cells were observed in the PVA/rhCOLIII-TA group compared to the other two groups (GLUT-PP and PVA-TA).…”

Combination of gene/protein and metabolite multiomics to reveal biomarkers of nickel ion cytotoxicity and the underlying mechanism

“…In previous studies, our research group utilized transcriptomic sequencing and iTRAQ-based proteomic techniques to investigate the gene and protein expression profiles of L929 cells treated with 100 and 200 μM Ni 2+ for 12, 24 and 48 h. Differentially expressed genes and proteins were identified in each experimental group [ 1 , 25 ]. In this article, we further selected gene/protein pairs that exhibited consistent differential expression (upregulated or downregulated) within the same Ni 2+ -treated group.…”

“…Biomarkers have been applied for toxicity assessment of biomaterials due to their advantages, such as rapid detection, simplicity and short time requirements [ 1–3 ]. However, research on biomarkers for biomaterials is still in its early stages.…”

“…Therefore, the safety of Ni 2+ and its interaction mechanisms with cells have been a subject of high interest among researchers [ 16 , 17 ]. Some researchers have employed traditional molecular biology methods to study the cytotoxicity biomarkers of Ni 2+ [ 18 , 19 ], but as of now, there have only been reports on the use of gene chip/transcriptome sequencing and proteomics for screening [ 1 , 20–22 ].…”

“…Over the years, our research group has employed cDNA microarray/transcriptome sequencing, proteomics and metabolomics technologies to investigate the toxic effects of Ni 2+ on L929 cells [ 23–25 ]. We have published the only paper to date that utilizes the integrated screening of transcriptomics/proteomics data and validates the expression levels and function of target biomarkers to identify cytotoxicity biomarkers of Ni 2+ [ 1 ].…”