Sonic Hedgehog activation is implicated in diosgenin-induced megakaryocytic differentiation of human erythroleukemia cells

Lamia Ghezali; Bertrand Liagre; Youness Limami; Jean-Louis Beneytout; David Yannick Leger

doi:10.1371/journal.pone.0095016

Sonic Hedgehog activation is implicated in diosgenin-induced megakaryocytic differentiation of human erythroleukemia cells

PLoS One. 2014 Apr 16;9(4):e95016. doi: 10.1371/journal.pone.0095016. eCollection 2014.

Authors

Lamia Ghezali¹, Bertrand Liagre¹, Youness Limami¹, Jean-Louis Beneytout¹, David Yannick Leger¹

Affiliation

¹ Université de Limoges, FR 3503 GEIST, EA 1069 "Laboratoire de Chimie des Substances Naturelles", GDR CNRS 3049, Faculté de Pharmacie, Laboratoire de Biochimie et Biologie Moléculaire, Limoges, France.

Abstract

Differentiation therapy is a means to treat cancer and is induced by different agents with low toxicity and more specificity than traditional ones. Diosgenin, a plant steroid, is able to induce megakaryocytic differentiation or apoptosis in human HEL erythroleukemia cells in a dose-dependent manner. However, the exact mechanism by which diosgenin induces megakaryocytic differentiation has not been elucidated. In this study, we studied the involvement of Sonic Hedgehog in megakaryocytic differentiation induced by diosgenin in HEL cells. First, we showed that different elements of the Hedgehog pathway are expressed in our model by qRT-PCR. Then, we focused our interest on key elements in the Sonic Hedgehog pathway: Smoothened receptor, GLI transcription factor and the ligand Sonic Hedgehog. We showed that Smoothened and Sonic Hedgehog were overexpressed in disogenin-treated cells and that GLI transcription factors were activated. Then, we showed that SMO inhibition using siSMO or the GLI antagonist GANT-61, blocked megakaryocytic differentiation induced by diosgenin in HEL cells. Furthermore, we demonstrated that Sonic Hedgehog pathway inhibition led to inhibition of ERK1/2 activation, a major physiological pathway involved in megakaryocytic differentiation. In conclusion, our study reports, for the first time, a crucial role for the Sonic Hedgehog pathway in diosgenin-induced megakaryocytic differentiation in HEL cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blotting, Western
Cell Differentiation / drug effects*
Cell Differentiation / genetics
Cell Line, Tumor
Diosgenin / pharmacology*
Gene Expression Regulation, Leukemic / drug effects
Hedgehog Proteins / genetics*
Hedgehog Proteins / metabolism
Humans
Leukemia, Erythroblastic, Acute / genetics
Leukemia, Erythroblastic, Acute / metabolism
Leukemia, Erythroblastic, Acute / pathology
Megakaryocytes / drug effects*
Megakaryocytes / metabolism
Mitogen-Activated Protein Kinase 1 / metabolism
Mitogen-Activated Protein Kinase 3 / metabolism
Phosphorylation / drug effects
Pyridines / pharmacology
Pyrimidines / pharmacology
RNA Interference
Receptors, G-Protein-Coupled / genetics
Receptors, G-Protein-Coupled / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Signal Transduction / drug effects
Signal Transduction / genetics
Smoothened Receptor
Time Factors
Transcription Factors / genetics
Transcription Factors / metabolism
Zinc Finger Protein GLI1

Substances

GANT 61
GLI1 protein, human
Hedgehog Proteins
Pyridines
Pyrimidines
Receptors, G-Protein-Coupled
SHH protein, human
SMO protein, human
Smoothened Receptor
Transcription Factors
Zinc Finger Protein GLI1
MAPK1 protein, human
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Diosgenin

Grants and funding

This research was supported by grants from the French Ministry of Education and Research and from the Conseil Régional du Limousin. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.