Transcriptomic investigation of the effects of TDCPP on PC12 and GC2 cells with experimental validation
Xi Zhang1, Qifu Zhang1, Yaohui Shan1
1Institute of Toxicology, College of Preventive Medicine, Third Military Medical University (Army Medical University), Chongqing 400038, China.
Abstract:
TDCPP is a flame retardant which has nervous and reproductive toxicity. Although there is a close association between nervous and reproductive system, the exact toxic mechanism of TDCPP in these systems is still seldom, especially in a genome scale. In this study, we explored the transcriptomic landscape of TDCPP in PC12 and GC2 cells using RNAseq method. A total of 465 co-differential expressed genes were found. These genes were mainly enriched in extra-cellular matrix, cell adhesion, cell cycle arrest, oxidoreductase activity GO terms, and PI3K/AKT, focal adhesion, ECM-receptor interaction KEGG pathways. Hub genes (ANXA1, COL27A1, GAS6, GNB4 and THBS1) were extracted using STRING and confirmed by qPCR experiment. Vimentin, HSPA5 and Caspase3 were proved to be responsible to TDCPP in GC2 and PC12 cells. Knockdown assay in PC12 cells showed that these hub genes could also affect the protein expression of vimentin, HSPA5 and Caspase3. In summary, TDCPP might exert its toxic effect through disturbing focal adhesion, ECM-receptor interaction and PI3K/Akt pathways. One of the mechanisms could be influence on the cytoskeleton (vimentin), ER stress (HSPA5) and apoptosis (Caspase3). The sequence data in this study might be a useful resource for future TDCPP related researches.
Insights
Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) exposure impacts nervous and reproductive systems. This study reveals TDCPP disrupts cellular pathways, affecting cytoskeleton, ER stress, and apoptosis, offering insights into its toxicity mechanisms.
Area of Science:
- Toxicology
- Genomics
- Cell Biology
Background:
- Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) is a flame retardant with known nervous and reproductive toxicity.
- The precise molecular mechanisms underlying TDCPP's toxicity, particularly at the genomic level, remain largely unexplored.
Purpose of the Study:
- To investigate the transcriptomic effects of TDCPP exposure in neuronal (PC12) and reproductive (GC2) cell lines.
- To identify key genes and pathways involved in TDCPP-induced toxicity.
Main Methods:
- RNA sequencing (RNAseq) was employed to analyze gene expression profiles in PC12 and GC2 cells treated with TDCPP.
- Bioinformatic analyses, including gene ontology (GO) and KEGG pathway enrichment, were performed.
- Hub genes were identified using STRING and validated by quantitative PCR (qPCR).
- Knockdown assays were conducted to confirm the role of identified genes.
Main Results:
- A total of 465 differentially expressed genes were identified.
- Enriched GO terms included extracellular matrix, cell adhesion, cell cycle arrest, and oxidoreductase activity.
- KEGG pathways implicated were PI3K/AKT, focal adhesion, and ECM-receptor interaction.
- Key genes such as ANXA1, COL27A1, GAS6, GNB4, and THBS1 were identified as potential mediators of TDCPP toxicity.
- Vimentin, HSPA5, and Caspase3 were confirmed as critical effectors in both cell types.
Conclusions:
- TDCPP toxicity may involve the disruption of focal adhesion, ECM-receptor interaction, and PI3K/Akt signaling pathways.
- Mechanisms include impacts on the cytoskeleton (vimentin), endoplasmic reticulum stress (HSPA5), and apoptosis (Caspase3).
- The generated transcriptomic data provides a valuable resource for future research on TDCPP toxicity.
