Transcriptional and translational-uncoupling in regulation of the CXCL12 and its receptors CXCR4, 7 in THP-1 monocytes and macrophages

Abstract

Introduction

The chemokine CXCL12 and its receptors CXCR4 and 7 play crucial roles in the immune system. In the present study, regulation of this pathway was further examined using the in-vitro model of undifferentiated human THP-1 monocytes (u-THP-1) and phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 macrophages (d-THP-1), to assess the effects of differentiation and the TLR4 ligand lipopolysaccharide (LPS) on the pathway.

Methods/Results

Differentiation did not affect the CXCR4, 7 mRNA levels. Interestingly, the CXCL12 and CXCR7 proteins but not CXCR4 were found to be up-regulated during differentiation. LPS, through CD14-dependent pathway, induced CXCL12 and CXCR4, 7 mRNA levels to a greater magnitude in d- than u-THP-1. The induction effect on CXCL12 stimulated by LPS was confirmed using ELISA. Increased migration of u-THP-1 was observed using conditioned medium from LPS-treated d-THP-1. Additionally, d-THP-1, although expressed higher CXCR7 protein levels, failed to migrate toward CXCL12. In contrast, LPS did not affect CXCR4, 7 protein levels.

Conclusion

Hence, this study indicated that CXCL12, CXCR4, and CXCR7 were differentially expressed and regulated in u-THP-1 and d-THP-1 cells in response to external stimuli. Importantly, we reported here a novel observation that uncoupling exists between transcriptional and translational regulation of CXCR4, 7 expressions by differentiation and TLR stimuli.

CXCL12 and CXCR4, 7 are differentially expressed and regulated in u-THP-1 and d-THP-1 cells in response to external stimuli. A disconnect between transcriptional and translational regulation of CXCR4, 7 expressions by differentiation and TLR stimuli exists.