Identification of the role of toxin B in the virulence of Clostridioides difficile based on integrated bioinformatics an
- PDF / 5,353,296 Bytes
- 13 Pages / 595.276 x 790.866 pts Page_size
- 3 Downloads / 178 Views
ORIGINAL ARTICLE
Identification of the role of toxin B in the virulence of Clostridioides difficile based on integrated bioinformatics analyses Yan Gao 1 & Weihu Gao 2 & Jingwei Cheng 1 & Liyan Ma 1 & Jianrong Su 1 Received: 8 October 2019 / Revised: 3 March 2020 / Accepted: 13 April 2020 # Springer Nature Switzerland AG 2020
Abstract Purpose Clostridioides difficile toxin B (TcdB) plays a critical role in C. difficile infection (CDI), a common and costly healthcare-associated disease. The aim of the current study was to explore the intracellular and potent systemic effects of TcdB on human colon epithelial cells utilizing Gene Expression Omnibus and bioinformatic methods. Methods Two datasets (GSE63880 and GSE29008) were collected to extract data components of mRNA of TcdB-treated human colon epithelial cells; “limma” package of “R” software was used to screen the differential genes, and “pheatmap” package was applied to construct heat maps for the differential genes; Metascape website was utilized for protein–protein interaction network and Molecular Complex Detection analysis, and Genome Ontology (GO) was used to analyze the selected differential genes. Quantitative real-time PCR (qRT-PCR) and Western blot were performed to validate the expression of hub genes. Results GO terms involved in DNA replication and cell cycle were identified significantly enriched in TcdB-treated human colon epithelial cells. Moreover, the decreased expression of DNA replication–related genes, MCM complex, and CDC45 in C. difficile (TcdA−/TcdB+)-infected Caco-2 cells were validated via qRT-PCR and Western blot assays. Conclusions In conclusion, the integrated analysis of different gene expression datasets allowed us to identify a set of genes and GO terms underlying the mechanisms of CDI induced by TcdB. It would aid in understanding of the molecular mechanisms underlying TcdB-exposed colon epithelial cells and provide the basis for developing diagnosis biomarkers, treatment, and prevention strategies. Keywords Clostridioides difficile . TcdB . Bioinformatics . CDC45 . MCM complex
Introduction Clostridioides difficile, a gram-positive and spore-forming bacterium, has been recognized as the most common and costly healthcare-associated pathogenic bacteria since it was first identified (Hall and O'toole 1935). Specifically, infection with a toxigenic strain of C. difficile leads to a range of clinical symptoms, from diarrhea and cramping in mild diseases to pseudomembranous colitis and even death in severe cases. The emergence of hypervirulent isolates of C. difficile, belonging to the BI/NAP1/027 strain, highlighted the severity
* Jianrong Su [email protected] 1
Department of Clinical Laboratory Diagnostics, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
2
Clinic School of Medicine, Anhui Medical University, Hefei 230032, Anhui, China
of C. difficile infection (CDI). An estimate showed that nearly half a million CDI cases and approximately 29,000 deaths occurred annually in the USA (Lessa et
Data Loading...
