title: Integration of genomic and proteomic data to identify candidate genes in HT-29 cells after incubation with Bifidobacterium bifidum ATCC 29521.
abstract: As the predominant group inhabiting the human gastrointestinal tract, bifidobacteria play a vital role in human nutrition, therapeutics, and health by shaping and maintaining the gut ecosystem, reducing blood cholesterol, and promoting the supply of nutrients. The interaction between bacterial cells and human intestinal epithelial cell lines has been studied for decades in an attempt to understand the mechanisms of action. These studies, however, have been limited by lack of genomic and proteomic database to aid in achieving comprehensive understanding of these mechanisms at molecular levels. Microarray data (GSE: 74119) coupled with isobaric tags for relative and absolute quantitation (iTRAQ) were performed to detect differentially expressed genes and proteins in HT-29 cells after incubation with Bifidobacterium bifidum. Real-time quantitative PCR, gene ontology, and Kyoto Encyclopedia of Genes and Genomes analyses were further conducted for mRNA validation, functional annotation, and pathway identification, respectively. According to the results of microarray, 1,717 differentially expressed genes, including 1,693 upregulated and 24 downregulated genes, were selected and classified by the gene ontology database. The iTRAQ analysis identified 43 differentially expressed proteins, where 29 proteins were upregulated and 14 proteins were downregulated. Eighty-two candidate genes showing consistent differences with microarray and iTRAQ were further validated in HT-29 and Caco-2 cells by real-time quantitative PCR. Nine of the top genes showing interesting results with high confidence were further investigated in vivo in mice intestine samples. Integration of genomic and proteomic data provides an approach to identify candidate genes that are more likely to function in ubiquitin-mediated proteolysis, positive regulation of apoptosis, membrane proteins, and transferase catalysis. These findings might contribute to our understanding of molecular mechanisms regulating the interaction between probiotics and intestinal epithelial cell lines.
authors.affiliation: State Key Laboratory of Food Science and Engineering, Nanchang University, Nanchang, Jiangxi 330047, P. R. China.
authors.forename: Bao-Gui
authors.initials: BG
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Food Science and Engineering, Nanchang University, Nanchang, Jiangxi 330047, P. R. China.
authors.forename: Yaoping
authors.initials: Y
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Food Science and Engineering, Nanchang University, Nanchang, Jiangxi 330047, P. R. China; Jiangxi University of Traditional Chinese Medicine, Nanchang, 330004, P.R. China.
authors.forename: Liang
authors.initials: L
authors.lastname: Qiu
authors.affiliation: Food and Nutritional Science, School of Biological Sciences, The University of Hong Kong, Hong Kong.
authors.forename: Nagendra P
authors.initials: NP
authors.lastname: Shah
authors.affiliation: State Key Laboratory of Food Science and Engineering, Nanchang University, Nanchang, Jiangxi 330047, P. R. China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Food Science and Engineering, Nanchang University, Nanchang, Jiangxi 330047, P. R. China. Electronic address: weihua@ncu.edu.cn.
authors.forename: Hua
authors.initials: H
authors.lastname: Wei
mesh_terms: D000818:Animals; D000069985:Bifidobacterium bifidum / Q000254:growth & development*; D018938:Caco-2 Cells; D030541:Databases, Genetic; D015536:Down-Regulation; D004847:Epithelial Cells / Q000378:metabolism* / Q000382:microbiology; D000069196:Gastrointestinal Microbiome; D063990:Gene Ontology; D019073:HT29 Cells; D006801:Humans; D007422:Intestines / Q000166:cytology / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D019936:Probiotics; D020543:Proteome; D040901:Proteomics; D015203:Reproducibility of Results; D015854:Up-Regulation
chemical_list: D020543:Proteome
keywords: Bifidobacterium bifidum; HT-29 cells; iTRAQ; microarray