阪崎肠杆菌培养上清蛋白引起HepG2肝细胞的基因表达变化

doi:10.3969/j.issn.1004-616x.2018.04.003

Abstract

Abstract: OBJECTIVE: To investigate differential gene expression of supernatant proteins from Cronobacter sakazakii on cytopathy of HepG2 cells and to understand molecular interactions in pathopoiesis. METHODS: The CAYE culture medium was used to culture Enterobacter cloacae,and supernatant proteins were isolated and purified by the ammonium sulfate precipitation method. The supernatant proteins were added to the 1×10⁶/mL HepG2 hepatocyte culture at a final concentration of 1.6 μg/mL as the experimental group and distilled water was added as the control group. After 8 h of treatment,total RNA was extracted,the cDNA library was constructed and were sequenced using Illumina. The difference in gene expression between the two groups was analyzed. The function of differentially expressed genes was analyzed by the Gene Ontology (GO) enrichment analysis and Pathway analysis. Differential expression of RPS18 and RFC4 genes were validated using qPCR. RESULTS: High throughput sequencing results showed that there were 5 120 differentially expressed genes:2 155 were up-regulated and 2 965 were down-regulated. GO enrichment analyses showed that the differentially expressed genes were mainly involved in several biological processes,including mitosis,caryomitosis,replication,etc. Pathway analysis showed that the differentially expressed genes were mainly involved in DNA replication,protein transport,and arginine and proline metabolism. The results of qPCR and high-throughput sequencing were consistent. CONCLUSION: The supernatant proteins of Cronobacter sakazakii caused HepG2 liver cell damage which might be related to changes of ribosome and/or DNA replication,and repair with differential expression of RPS18 and RFC4 genes.

Key words: Cronobacter sakazakii, supernatant, HepG2 cell, high-throughput sequencing

CLC Number:

R155.5

YOU Shuping, REN Lisong, MA Long, HE Sun, CHEN Zeliang, WANG Rong, LIU tao. Differential gene expression of supernatant proteins from Cronobacter sakazakii on cytopathy of HepG2 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(4): 258-262.

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Differential gene expression of supernatant proteins from Cronobacter sakazakii on cytopathy of HepG2 cells

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