Comprehensive analysis of differentially expressed microRNAs and mRNAs in MDBK cells expressing bovine papillomavirus E5 oncogene

Feng Pang; Zhen Chen; Chengqiang Wang; Mengmeng Zhang; Zhenxing Zhang; Xiaohong Yang; Yiying Zheng; Ang Liu; Yiwen Cheng; Jie Chen; Bin Li; Li Du; Fengyang Wang

doi:10.7717/peerj.8098

Comprehensive analysis of differentially expressed microRNAs and mRNAs in MDBK cells expressing bovine papillomavirus E5 oncogene

PeerJ. 2019 Nov 22:7:e8098. doi: 10.7717/peerj.8098. eCollection 2019.

Authors

Feng Pang^#¹, Zhen Chen^#¹, Chengqiang Wang¹, Mengmeng Zhang¹, Zhenxing Zhang¹, Xiaohong Yang¹, Yiying Zheng¹, Ang Liu¹, Yiwen Cheng¹, Jie Chen¹, Bin Li¹, Li Du¹, Fengyang Wang¹

Affiliation

¹ College of Animal Science and Technology, Hainan University, Hainan Key Lab of Tropical Animal Reproduction & Breeding and Epidemic Disease Research, Hainan University, Haikou, China.

^# Contributed equally.

Abstract

Delta bovine papillomaviruses (δBPVs) causes fibropapillomas or bladder cancer in cattle. E5 is the major oncogene of δBPVs; however, the influence that E5 oncogene has on host microRNA (miRNA) and mRNA expression profiles remains little elucidated. In the present study, small RNA sequencing and RNA sequencing were used to explore alterations in miRNAs and mRNAs in E5 over-expressing Madin-Darby bovine kidney (MDBK) cells compared with controls. In total, 77 miRNAs (including 30 bovine-derived miRNAs) and 223 genes were differentially expressed (DE) following E5 overexpression. The dysregulated genes were mainly involved in metabolic and biosynthetic processes. We constructed a potential miRNA-gene regulatory network from the differentially expressed genes (DEGs) and DE miRNAs. Finally, 22 DEGs and nine DE miRNAs were selected for RT-qPCR validation. Of these, downregulation of six miRNAs, bta-miR-34c, bta-miR-122, bta-miR-195, bta-miR-449b, bta-miR-2425-5p, and bta-miR-2428-3p were confirmed; In addition, upregulation of 16 genes, ACSS2, DDIT4, INHBE, INSIG1, PNRC1, PSAT1, PSPH, PYCR1, SC4MOL, SLC34A2, SCD, SPARC, IDI1, PCK2, HMGCS1, and SMIM14 and downregulation of two genes, BATF3 and WFDC2 were confirmed. Specially, bta-miR-34c and bta-miR-449b potentially regulated PYCR1 and DDIT4, which were involved in cancer progression and angiogenesis. Our study presented for the first time the comprehensive miRNA and mRNA alterations in MDBK cells expressing the BPV E5 oncogene, providing new insights into the tumorigenesis induced by BPV E5.

Keywords: Bovine papillomavirus; E5; RNA-seq; Target genes; miRNA-gene network; microRNAs.

Grants and funding

This work was financially supported by Hainan Natural Science Foundation Innovation Research Team Project (2017CXTD003) and Projects for the central government to guide local scientific and technological development (ZY2019HN0903). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.