Osteopontin (OPN/SPP1), a Mediator of Tumor Progression, Is Regulated by the Mesenchymal Transcription Factor Slug/SNAI2 in Colorectal Cancer (CRC)

SIMPLE SUMMARY: Expression of the transcription factor Slug/SNAI2 is associated with the epithelial–mesenchymal transition (EMT) and is correlated with poorer disease-free survival in colorectal cancer (CRC). In order to decipher the basis for the Slug-mediated aggressive phenotype, we conducted RNAseq experiments with a panel of HT-29 CRC cells expressing different levels of Slug, both in vitro and in tumor models. Osteopontin (OPN), a mediator associated with tumor progression in different tumor types, was among the top upregulated genes in both cells and tumors and was the most overexpressed gene coding for a secreted protein. We further show that Slug is a direct regulator of osteopontin via binding to the OPN promoter. Interestingly, Slug expression and osteopontin secretion were correlated in vitro, as well as in tumor models, suggesting that liquid biopsies may be useful in estimating the aggressiveness phenotype of the tumor. ABSTRACT: In colorectal cancer (CRC), disease-related death is closely linked to tumor aggressiveness and metastasis. Gene expression profiling of patient tumors has suggested that a more mesenchymal phenotype, present in about one-fourth of all patients, is associated with increased aggressiveness. Accordingly, the mesenchymal transcription factor Slug/SNAI2 has been associated with decreased disease-free survival. To decipher the basis for the Slug-mediated phenotype, we conducted RNAseq experiments with a panel of HT-29 CRC cells expressing different levels of Slug, both in vitro and in tumor models. The results show that osteopontin, a secreted pleotropic protein involved in multiple steps of colorectal cancer progression, was highly upregulated by Slug in vitro, as well as in vivo. We further show that Slug is a direct regulator of osteopontin at the promoter level. The levels of secreted osteopontin were correlated with Slug expression, thereby linking the tumor phenotype to a biomarker available by liquid biopsies. The results also suggest that osteopontin neutralization may attenuate at least some of the Slug-mediated functions.