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Thromb Haemost 2016; 116(05): 931-940
DOI: 10.1160/TH15-09-0749
Cellular Haemostasis and Platelets
Schattauer GmbH

Nuclear factor-κB regulates expression of platelet phospholipase C-β2 (PLCB2)

Guangfen Mao
1   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Jianguo Jin
1   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
3   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Satya P. Kunapuli
1   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
3   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
A. Koneti Rao
1   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2   Department of Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
› Author Affiliations Financial support: This work was supported by grants from the National Institutes of Health R01HL109568, R01HL85422 and R01HL56724 (AKR), and R01HL93231 and R01HL118593 (SPK).
Further Information

Publication History

Received: 22 September 2016

Accepted after major revision: 10 July 2016

Publication Date:
30 November 2017 (online)

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Summary

Phospholipase C (PLC)-β2 (gene PLCB2) is a critical regulator of platelet responses upon activation. Mechanisms regulating of PLC-β2 expression in platelets/MKs are unknown. Our studies in a patient with platelet PLC-β2 deficiency revealed the PLCB2 coding sequence to be normal and decreased platelet PLC-β2 mRNA, suggesting a defect in transcriptional regulation. PLCB2 5’- upstream region of the patient revealed a heterozygous 13 bp deletion (-1645/-1633 bp) encompassing a consensus sequence for nuclear factor-κB (NF-κB). This was subsequently detected in three of 50 healthy subjects. To understand the mechanisms regulating PLC-β2, we studied the effect of this variation in the PLCB2. Gel-shift studies using nuclear extracts from human erythroleukaemia (HEL) cells or recombinant p65 showed NF-κB binding to oligonucleotide with NF-κB site; in luciferase reporter studies its deletion reduced PLCB2 promoter activity. PLCB2 expression was decreased by siRNA knockdown of NF-κB p65 subunit and increased by p65 overexpression. By immunoblotting platelet PLC-β2 in 17 healthy subjects correlated with p65 (r=0.76, p=0.0005). These studies provide the first evidence that NF-kB regulates MK/platelet PLC-β2 expression. This interaction is important because of the major role of PLC-β2 in platelet activation and of NF-κB in processes, including inflammation and atherosclerosis, where both are intimately involved.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Megakaryocytess - nuclear factor-κB - phospholipase C-β2 - platelet function defect - platelets

 

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