Department of Medical Biochemistry, Medical University of Lodz, Lodz, Poland.
Int J Mol Med. 2011 Nov;28(5):875-80. doi: 10.3892/ijmm.2011.748. Epub 2011 Jul 12.
A tight relationship has been revealed between cellular microRNAs (miRNAs) and the course of hepatitis C virus (HCV) replication in human hepatoma cells. Although the detection of the antigenomic HCV RNA strand in peripheral blood mononuclear cells (PBMCs) has provided evidence for viral replication in PBMCs, no reports have shown how miRNAs are affected upon HCV RNA synthesis in PBMCs. The aim of the present study was to assess if and how the expression levels of miRNA-155 and miRNA-196b in PBMCs are related to HCV replication in PBMCs of chronic hepatitis C (CHC) patients. Supporting analyses were performed to evaluate the expression of precursor pri-miR-155 (BIC) and Dicer protein. The genomic and antigenomic HCV RNA strands in PBMCs were detected by strand-specific qRT-PCR. The expression levels of miRNAs, BIC RNA and Dicer protein were assayed on PBMCs by qRT-PCR and Western blotting, respectively. miRNA-155 and miRNA-196b were detected in all studied PBMC samples, but their levels varied according to the presence of the antigenomic HCV RNA strand in PBMCs. Increased expression levels of miRNA-155 and miRNA-196b were associated with the presence of the antigenomic HCV RNA strand in PBMCs. In this group of patients higher frequency of BIC RNA and Dicer protein detection was also found. This study demonstrates that HCV RNA replication in PBMCs of CHC patients is connected with the increased and coordinated expression of miRNA-155 and miRNA-196b.
细胞 microRNAs（miRNAs）与丙型肝炎病毒（HCV）在人肝癌细胞中的复制过程之间存在紧密关系。尽管在外周血单核细胞（PBMCs）中检测到抗基因组 HCV RNA 链为 PBMCs 中的病毒复制提供了证据，但尚无报道表明 HCV RNA 在 PBMCs 中的合成如何影响 miRNAs。本研究旨在评估慢性丙型肝炎（CHC）患者 PBMCs 中的 miRNA-155 和 miRNA-196b 的表达水平是否与 HCV 在 PBMCs 中的复制有关。进行了支持性分析以评估 PBMCs 中 miRNA-155 的前体 pri-miR-155（BIC）和 Dicer 蛋白的表达。通过特异性 qRT-PCR 检测 PBMCs 中的基因组和抗基因组 HCV RNA 链。通过 qRT-PCR 和 Western blot 分别检测 PBMCs 中 miRNA、BIC RNA 和 Dicer 蛋白的表达水平。在所研究的 PBMC 样本中均检测到了 miRNA-155 和 miRNA-196b，但它们的水平根据 PBMCs 中抗基因组 HCV RNA 链的存在而有所不同。miRNA-155 和 miRNA-196b 的表达水平增加与 PBMCs 中抗基因组 HCV RNA 链的存在相关。在该组患者中，还发现了更高频率的 BIC RNA 和 Dicer 蛋白检测。本研究表明，CHC 患者 PBMCs 中的 HCV RNA 复制与 miRNA-155 和 miRNA-196b 的增加和协调表达有关。
