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Published on: November 30, 2016
[miRNA Expression Profile in Ileocecal Adenocarcinoma Cells Infected with Cryptosporidium]
Özlem Ulusan Bağcı1, Ayşe Caner2
1Katip Celebi University, Ataturk Training and Research Hospital, Laboratory of Microbiology, İzmir, Türkiye.
Abstract:
Cryptosporidium spp. is an opportunistic protozoan transmitted by fecal-oral route via oocysts. The agent may cause severe infection especially in individuals with suppressed immune system, due to its intracellular location and ability to cause auto-infection. MicroRNAs (miRNAs) are non-translated endogenous RNA molecules with an average of 22 nucleotides in length that regulate the expression of genes involved in important biological functions such as proliferation, differentiation, apoptosis and immune response. Recent studies have focused on the role of miRNAs in pathogenesis of infectious diseases and their potential to be used as biomarkers. The aim of this study was to determine the miRNA profile of human ileocecal adenocarcinoma (HCT-8) cells at 24 hours of infection with Cryptosporidium spp. In the study, the HCT-8 cell line was infected with Cryptosporidium spp. that were isolated from infected human stool samples and RNA was isolated from the cells 24 hours after infection. After this process, cDNA synthesis was performed and the expression of 95 human miRNA profiles were investigated by polymerase chain reaction (PCR) method. Fold changes of expression were determined by comparison with Cryptosporidium spp. uninfected cell lines. Sequence information of miRNAs and their target genes were performed via TargetScanHuman7.1 and miRDB websites, while gene ontology (GO) pathways of target genes were analyzed with the mirPath v.3 program. It was detected that the expression of 10 miRNAs were upregulated and 11 of them were downregulated compared with the control group. It was observed that, this 21 differentially expressed miRNAs were mainly associated with apoptosis, mitotic cell cycle, and immune response. Hsa-miR-612, hsa-miR-6763-5p, hsa-miR-188-5p, hsa-miR-664b-3p, hsa-miR-210-3p, hsa-let-7e-5p hsa-let-7b-3p, hsa-miR-4787-3p, hsa-miR-548ab, hsa-miR-3714 and hsamiR-4803 were found to be associated with apoptosis; and hsa-miR-612, hsa-miR-664b-3p, hsa-miR210-3p, hsa-let-7e-5p, hsa-let-7b-3p, hsa-miR-548ab, and hsa-miR4803 were found to be associated with mitotic cell cycle. The balance of proliferation and apoptosis is very significant in the development of infection and cancer. It is thought that determination of the effect of miRNAs on proliferation-apoptosis balance could provide information related to the etiopathogenesis and prognosis of infections, and on the role of microorganisms in carcinogenesis. In this study, 12 differentially expressed miRNAs were found to be associated with immune response. This may emphasize the role of miRNAs in the prevention and treatment of infections. It was concluded that, miRNAs could be used in the diagnosis, treatment and prevention of infections with the determination of miRNA's role in the infection mechanism as a result of the increasing number of studies.
Insights
This study investigated microRNAs (miRNAs) in human cells infected with Cryptosporidium, revealing significant changes in miRNA expression linked to apoptosis and immune response. These findings suggest miRNAs could be key biomarkers for diagnosing and treating parasitic infections.
Area of Science:
- Parasitology
- Molecular Biology
- Genomics
Background:
- Cryptosporidium spp. is an opportunistic protozoan pathogen causing severe infections, particularly in immunocompromised individuals.
- MicroRNAs (miRNAs) are crucial regulators of gene expression involved in cellular processes like apoptosis and immune response.
- Emerging research highlights miRNAs' potential as biomarkers in infectious diseases.
Purpose of the Study:
- To profile microRNA (miRNA) expression in human ileocecal adenocarcinoma (HCT-8) cells following Cryptosporidium spp. infection.
- To identify differentially expressed miRNAs and their associated biological pathways.
- To explore the role of miRNAs in the host-pathogen interaction and cellular responses.
Main Methods:
- Human ileocecal adenocarcinoma (HCT-8) cells were infected with Cryptosporidium spp.
- RNA was isolated 24 hours post-infection for cDNA synthesis.
- Polymerase chain reaction (PCR) was used to analyze the expression of 95 human miRNAs.
- Bioinformatic tools (TargetScanHuman7.1, miRDB, mirPath v.3) were used for target gene and pathway analysis.
Main Results:
- 21 differentially expressed miRNAs were identified: 10 upregulated and 11 downregulated.
- These miRNAs were significantly associated with apoptosis, mitotic cell cycle, and immune response pathways.
- Specific miRNAs like hsa-miR-612 and hsa-let-7e-5p were linked to apoptosis and cell cycle regulation.
Conclusions:
- The study identified key miRNAs involved in the host cellular response to Cryptosporidium infection.
- Differentially expressed miRNAs highlight the intricate relationship between infection, apoptosis, and immune modulation.
- These findings support the potential of miRNAs as diagnostic and therapeutic targets for Cryptosporidium infections.

