Ulusan Bağcı Özlem, Caner Ayşe
Katip Celebi University, Ataturk Training and Research Hospital, Laboratory of Microbiology, İzmir, Türkiye.
Ege University Institute of Health Sciences, Department of Basic Oncology, İzmir, Türkiye.
Mikrobiyol Bul. 2022 Jul;56(3):449-465. doi: 10.5578/mb.20229706.
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.
隐孢子虫属是一种通过卵囊经粪-口途径传播的机会性原生动物。由于其细胞内定位和引起自身感染的能力，该病原体尤其可在免疫系统受抑制的个体中引发严重感染。微小RNA（miRNA）是平均长度为22个核苷酸的非翻译内源性RNA分子，可调节参与增殖、分化、凋亡和免疫反应等重要生物学功能的基因表达。最近的研究集中在miRNA在传染病发病机制中的作用及其作为生物标志物的潜力。本研究的目的是确定隐孢子虫属感染24小时后人回盲部腺癌（HCT-8）细胞的miRNA谱。在该研究中，用从感染的人类粪便样本中分离出的隐孢子虫属感染HCT-8细胞系，并在感染后24小时从细胞中分离RNA。在此过程之后，进行cDNA合成，并通过聚合酶链反应（PCR）方法研究95种人类miRNA谱的表达。通过与未感染隐孢子虫属的细胞系比较来确定表达的倍数变化。miRNA及其靶基因的序列信息通过TargetScanHuman7.1和miRDB网站获得，而靶基因的基因本体（GO）途径则用mirPath v.3程序进行分析。与对照组相比，检测到10种miRNA的表达上调，11种miRNA的表达下调。观察到，这21种差异表达的miRNA主要与凋亡、有丝分裂细胞周期和免疫反应相关。发现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和hsa-miR-4803与凋亡相关；并且发现hsa-miR-612、hsa-miR-664b-3p、hsa-miR210-3p、hsa-let-7e-5p、hsa-let-7b-3p、hsa-miR-548ab和hsa-miR4803与有丝分裂细胞周期相关。增殖和凋亡的平衡在感染和癌症的发展中非常重要。认为确定miRNA对增殖-凋亡平衡的影响可为感染的病因发病机制和预后以及微生物在致癌作用中的作用提供相关信息。在本研究中，发现12种差异表达的miRNA与免疫反应相关。这可能强调了miRNA在感染预防和治疗中的作用。得出的结论是，随着研究数量的增加，通过确定miRNA在感染机制中的作用，miRNA可用于感染的诊断、治疗和预防。
