Whole-Transcriptome Analysis of Three Human Cell Lines Stably Infected with Bovine Leukemia Virus (BLV) Reveals Novel Apoptosis-Associated Factors

Bovine leukemia virus (BLV), a major cause of B-cell lymphoma in cattle worldwide, has been linked to human breast cancer. Here, we performed comparative whole-transcriptome analysis of single clones of human epithelial 293T, breast cancer MCF7, and cervical cancer HeLa cells, stably infected with BLV, versus uninfected controls using RNA-sequencing technology. Differential expression analysis revealed 2050, 1314, and 2772 differentially expressed genes (DEGs) between BLV-infected 293T, MCF7, and HeLa cells and controls, respectively. Most DEGs were upregulated in BLV-infected 293T (76.5%) and MCF7 (69.1%) cells but downregulated in HeLa cells (62.0%). Functional enrichment analyses revealed enrichment of “Gene expression” and “Membrane Trafficking” pathways across all cell lines, and that of “Apoptosis” and “Axon guidance” pathways uniquely in 293T and MCF7 cells. Twenty genes linked to apoptosis of 293T and MCF7 cells were identified, and expression of 11 of these was confirmed using quantitative real-time PCR. The expression of EFEMP1 and LXN, which showed the greatest fold change, was validated: EFEMP1 knockdown and LXN overexpression inhibited cell proliferation, induced apoptosis, and altered growth morphology of 293T and MCF7 cells. This is the first transcriptome profiling of human cells during BLV latency, which should help understand the behavior of BLV.