Gene expression change in human dental pulp cells exposed to a low-level toxic concentration of triethylene glycol dimethacrylate: an RNA-seq analysis.
Dental composite resin restoration for defective tooth may lead unpolymerized resin monomers to be leached into dental pulp tissue. The aim of this study was to investigate the early gene expression change over time of human dental pulp cells (HDPCs) treated with a low-level toxic concentration of Triethylene Glycol Dimethacrylate (TEGDMA), a common dental resin monomer, by adopting the novel high-throughput transcriptome analysis of RNA-seq. The low-level toxic concentration of TEGDMA was determined through MTT assays with serially diluted concentrations. After the HDPCs were exposed to TEGDMA for 6, 12, 24 or 48 hr, the total RNA of the samples was prepared for RNA-seq. qRT-PCR for several genes was performed for validation of RNA-seq results. In the treated group, 1280 genes were differentially expressed compared with the control group. Five patterns of time-series gene expression profiles were identified through k-means clustering analysis. Angiogenesis, cell adhesion and migration, extracellular matrix organization, response to extracellular stimulus, inflammatory response and mineralization-related process were major gene ontology terms in functional annotation clustering. HMOX1, OSGIN1, SMN2, SRXN1 AKR1C1, SPP1 and TOMM40L were highly up-regulated genes, and WRAP53 and CCL2 were highly down-regulated genes over time. qRT-PCR for several genes exhibited a high level of agreement with RNA-seq. TEGDMA induced the HDPCs to show massive and dynamic gene expression changes over time. The previously suggested toxic mechanism of TEGDMA was not only verified, but new genes whose functions have yet to be determined were also found.
Department of Conservative Dentistry, School of Dentistry, Kyung Hee University, Seoul, Korea.
Sung-Geun
SG
Cho
Jin-Woo
JW
Lee
Jung Sun
JS
Heo
Sun-Young
SY
Kim
D000180:Acrylic Resins / Q000737:chemistry; D043925:Angiogenesis Inducing Agents / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D003188:Composite Resins / Q000737:chemistry; D003782:Dental Pulp / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D015536:Down-Regulation; D015870:Gene Expression; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D005810:Multigene Family; D011092:Polyethylene Glycols / Q000633:toxicity*; D011109:Polymethacrylic Acids / Q000633:toxicity*; D011140:Polyurethanes / Q000737:chemistry; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017423:Sequence Analysis, RNA; D059467:Transcriptome; D015854:Up-Regulation
D000180:Acrylic Resins; D043925:Angiogenesis Inducing Agents; C029012:Composite Dental Resin; D003188:Composite Resins; D011109:Polymethacrylic Acids; D011140:Polyurethanes; C020946:triethylene glycol dimethacrylate; D011092:Polyethylene Glycols