MicroRNA expression profile of HCT-8 cells in the early phase of Cryptosporidium parvum infection

Abstract

BackgroundCryptosporidium parvum is an important zoonotic parasitic disease worldwide, but the molecular mechanisms of the host–parasite interaction are not fully understood. Noncoding microRNAs (miRNAs) are considered key regulators of parasitic diseases. Therefore, we used microarray, qPCR, and bioinformatic analyses to investigate the intestinal epithelial miRNA expression profile after Cryptosporidium parvum infection.ResultsTwenty miRNAs were differentially expressed after infection (four upregulated and 16 downregulated). Further analysis of the differentially expressed miRNAs revealed that many important cellular responses were triggered by Cryptosporidium parvum infection, including cell apoptosis and the inflammatory and immune responses.ConclusionsThis study demonstrates for the first time that the miRNA expression profile of human intestinal epithelium cells is altered by C. parvum infection. This dysregulation of miRNA expression may contribute to the regulation of host biological processes in response to C. parvum infection, including cell apoptosis and the immune responses. These results provide new insight into the regulatory mechanisms of host miRNAs during cryptosporidiosis, which may offer potential targets for future C. parvum control strategies.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-5410-6) contains supplementary material, which is available to authorized users.

Cryptosporidium parvum upregulates miR-942-5p expression in HCT-8 cells via TLR2/TLR4-NF-κB signaling

“…Our previous study found that miR-942-5p was strongly upregulated in HCT-8 cells following C. parvum infection, compared with uninfected cells [18]. In this study, qPCR assays revealed that the transcription of the miR-942 gene was significantly decreased after blocking either TLR2 or TLR4 (F (2, 23) = 58.32, P = 10 −9 by one-way ANOVA: test versus control group for Fig.…”

“…As previously described, oocysts were excysted in 0.25% trypsin and 0.75% sodium taurocholate for 1 h with mixing every 5 min, followed by incubation at room temperature for 30 min [19,20]. HCT-8 human ileocecal adenocarcinoma cells (American Type Culture Collection, Manassas, VA) were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 4 mmol/l l-glutamine, 100 U/ml penicillin, and 100 U/ml streptomycin at 37 °C in a humidified 5% CO 2 incubator [18]. Cell monolayers in 24-well cell culture dishes were inoculated with 2.5 × 10 6 purified sporozoites per well in DMEM.…”

“…Our previous study of the miRNAs expressed in HCT-8 cells infected with C. parvum found that miR-942-5p, miR-181d, miR-3976, miR-18b-3p, miR-34b-5p and miR-3591-3p may regulate apoptosis in the early phase of infection [18]. This study investigated the upregulation of miR-942-5p in cultured HCT-8 human ileocecal adenocarcinoma cells following activation of the TLR2/TLR4-NF-κB signaling pathway by C. parvum.…”

Human microRNAs in host–parasite interaction: a review

“…Specifically, C. parvum infection downregulates the expression of miR-98 and let-7 family to induce the suppressor of cytokine signaling (SOCS4) proteins that are negative regulators of cytokine signaling (Hu et al 2010 ; Sato et al 2017 ). Recently, Wang and collaborators ( 2019 ) reported for the first time the miRNA expression profile of human intestinal epithelial cells infected with C. parvum and they evidenced that most miRNAs were not significantly differentially expressed in the infected HCT-8 cells as compared to uninfected cells. Nonetheless, they reported that the miR-34b-5p, miR-18b-3p, miR-3976, and miR-3591-3p were downregulated after C. parvum infection and those have been associated with the regulation of both apoptotic processes and epithelial immune responses, targeting genes, such as ELAVL1, RAB10, RAB14, ELK4, SOS2, TAB2, DAXX, fibroblast growth factor 14 (FGF14), and MAPK3.…”

The Influence of Selected Gastrointestinal Parasites on Apoptosis in Intestinal Epithelial Cells

“…Additionally, the application of epidermal growth factor (EGF) to the cell culture resulted in the inhibition of apoptotic processes induced by Cryptosporidium [68]. Furthermore, it was found that the process is extremely complex and determined by numerous factors, among others, by the research model, duration of infection, and the developmental stage of the parasite [69][70][71][72][73][74][75][76][77].…”