HSV-2-encoded miRNA-H4 Regulates Cell Cycle Progression and Act-D-induced Apoptosis in HeLa Cells by Targeting CDKL2 and CDKN2A
- 39 Downloads
MicroRNAs (miRNAs) encoded by latency-associated transcript are associated with both latent and acute stages of herpes simplex virus 2 (HSV-2) infection. In this study, miRNA-H4-5p and miRNA-H4-3p were ectopically expressed in HeLa cells to explore potential cellular targets of viral miRNAs and demonstrate their potential biological functions. The results showed that miRNA-H4-5p could reverse apoptosis induced by actinomycin D (Act-D) and promote cell cycle progression, but miRNA-H4-3p had no such obvious functions. Bioinformatics analysis, luciferase report assay, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and Western blotting demonstrated that miRNA-H4-5p could bind to the 3′-untranslated region (UTR) of cyclin-dependent kinase inhibitor 2A (CDKN2A) and cyclin-dependent kinase-like 2 (CDKL2) to negatively regulate their expression. We verified that these two targeted genes were associated with cell apoptosis and cell cycle. Furthermore, in HeLa cells infected with HSV-2, we detected significantly reduced expression of CDKN2A and CDKL2 and demonstrated the negative regulation effect of miRNA-H4-5p on these two target genes. Our findings show that viral miRNAs play a vital role in regulating the expression of the host’s cellular genes that participate in cell apoptosis and progression to reshape the cellular environment in response to HSV-2 infection, providing further information on the roles of encoded herpesvirus miRNAs in pathogen–host interaction.
KeywordsHerpes simplex virus 2 (HSV-2) MicroRNAs (miRNAs) Anti-apoptosis Cell cycle progression Pathogen–host interaction
This work was supported by the National Natural Science Foundation of China (81371749) and (81171511).
HY and JF designed the study; YZ, JY and YL performed the experiments; YZ and JY analyzed the data; YZ and YL wrote the manuscript; HY and JF finalized the manuscript. All the authors approved the final manuscript.
Compliance with Ethics standards
Conflict of interest
The authors declare that they have no conflict of interest.
Animal and Human Rights Statement
The authors declare that they have no conflict of interest. This article does not contain any studies with human or animal subjects performed by any of the authors.
- Gomi H, Sassa T, Thompson RF, Itohara S (2010) Involvement of cyclin-dependent kinase-like 2 in cognitive function required for contextual and spatial learning in mice. Front Behav Neurosci 4:17Google Scholar
- Journey LJ, Goldstein MN (1961) Electron microscope studies on HeLa cell lines sensitive and resistant to actinomycin D. Cancer Res 21:929–932Google Scholar
- Li L, Liu C, Amato RJ, Chang JT, Du G, Li W (2014) CDKL2 promotes epithelial-mesenchymal transition and breast cancer progression. Oncotarget 5:10840–10853Google Scholar
- Lu DF, Wang YS, Li C, Wei GJ, Chen R, Dong DM, Yao M (2015) Actinomycin D inhibits cell proliferations and promotes apoptosis in osteosarcoma cells. Int J Clin Exp Med 8:1904–1911Google Scholar
- Pal A, Potjer TP, Thomsen SK, Ng HJ, Barrett A, Scharfmann R, James TJ, Bishop DT, Karpe F, Godsland IF, Vasen HF, Newton-Bishop J, Pijl H, McCarthy MI, Gloyn AL (2016) Loss-of-function mutations in the cell-cycle control gene CDKN2A impact on glucose homeostasis in humans. Diabetes 65:527–533CrossRefGoogle Scholar
- Tang S, Bertke AS, Patel A, Margolis TP, Krause PR (2011) Herpes simplex virus 2 microRNA miR-H6 is a novel latency-associated transcript-associated microRNA, but reduction of its expression does not influence the establishment of viral latency or the recurrence phenotype. J Virol 85:4501–4509CrossRefGoogle Scholar
- Whitley RJ (2015) Herpes simplex virus infections of the central nervous system. Continuum (Minneap Minn) 21:1704–1713Google Scholar