OBJECTIVE: To study the expression of Shh and Wnt5a genes in the limb buds of NIPBL fetal rats and the association of these two genes with Cornelia de Lange syndrome (CdLS). METHODS: A total of 72 NIPBL fetal rats were divided into an experimental group and a control group, with 36 rats in each group. The limb buds were collected from 12 fetal rats each on embryonic days 10, 11 and 12 (E10, E11 and E12) respectively. Real-time PCR and Western blot were used to measure the mRNA and protein expression of Shh and Wnt5a. RESULTS: The mRNA and protein expression of Shh and Wnt5a was detected in the limb buds on E10, E11 and E12, and the experimental group had significantly lower expression than the control group (P<0.01). The mRNA and protein expression of Shh and Wnt5a in limb buds was at a low level on E10, followed by an increase on E11 and a reduction on E12, and the expression on E12 was still lower than that on E10 (P<0.01). CONCLUSIONS The mRNA and protein expression of Shh and Wnt5a are consistent. The pathogenesis of CdLS may be associated with the low mRNA and protein expression of Shh and Wnt5a inhibited by the low expression of NIPBL gene.
Animals ; De Lange Syndrome ; Hedgehog Proteins ; Mutation ; Phenotype ; Proteins ; RNA, Messenger ; Rats ; Wnt-5a Protein

RNA binding protein (RBP) plays a key role in gene regulation and participate in RNA translation, modification, splicing, transport and other important biological processes. Studies have shown that abnormal expression of RBP is associated with a variety of diseases. The Musashi (Msi) family of mammals is an evolutionarily conserved and powerful RBP, whose members Msi1 and Msi2 play important roles in the regulation of stem cell activity and tumor development. The Msi family members regulate a variety of biological processes by binding and regulating mRNA translation, stability and downstream cell signaling pathways, and among them, Msi2 is closely related to embryonic growth and development, maintenance of tumor stem cells and development of hematological tumors. Accumulating evidence has shown that Msi2 also plays a crucial role in the development of solid tumors, mainly by affecting the proliferation, invasion, metastasis and drug resistance of tumors, involving Wnt/β-catenin, TGF-β/SMAD3, Akt/mTOR, JAK/STAT, Numb and their related signaling pathways (Notch, p53, and Hedgehog pathway). Preclinical studies of Msi2 gene as a therapeutic target for tumor have achieved preliminary results. This review summarizes the molecular structure, physiological function, role of Msi2 in the development and progression of various solid tumors and the signaling pathways involved.
Animals ; Hedgehog Proteins ; Mammals/metabolism* ; Neoplasms/genetics* ; Neoplastic Stem Cells ; RNA-Binding Proteins/metabolism* ; Signal Transduction

Sonic hedgehog (Shh) is a key signal regulatory factor in embryonic development. It is reported that Shh signaling plays important roles in prostatic duct differentiation and matrix-epithelium interaction, and thus regulates the development, growth and cell proliferation of the prostate. A disorder in Shh signaling will lead to the production and proliferation of tumor cells. An exploration into the mechanism of Shh signaling in the normal growth and diseased condition of the prostate will offer some significant ideas for the studies on the pathogenesy of prostate diseases.
Animals ; Hedgehog Proteins ; physiology ; Humans ; Male ; Prostate ; growth & development ; Signal Transduction ; physiology

OBJECTIVETo observe the role of the Hedgehog (Hh) genes in the proliferation of osteoblasts upon mechanical tensile strains.

METHODSPrimary osteoblasts harvested from newborn rat calvarial bone were subjected to 3% and 6% elongation of tensile stretches using Flexcell 4000 strain unit. The cultures were also treated with either recombinant N-terminals Sonic Hedgehog (N-Shh) or cyclopamine (cy), a Hh inhibitor or gadolinium (GdCl3), an inhibitor of stretch-activated channels. The proliferation of osteoblasts was quantified by cell counting, methyl thiazolyl tetrazolium (MTT) and cell cycle detection via flow cytometry. Statistical analysis was performed using SAS 8.0 software package.

RESULTSThe tensile strain, especially under 6% elongation, promoted osteoblast proliferation. Stretching force could also promote the proliferation even when the cells were treated with cy, but this effect was suppressed by GdCl3.

CONCLUSIONThe induced proliferation of osteoblasts by mechanical stretched is mediated at least in part by Indian Hedgehog (Ihh) signaling.

Drug resistance and relapse are the major challenge for current treatment of acute leukemia. It is critical for ultimately curing leukemia to overcome chemoresistance of leukemic stem cells (LSC) and to eradicate LSC. Recent studies have found that abnormal activated Hedgehog (HH) signaling pathway plays an important role in a wide variety of tumors and regulates multi-drug resistance of LSC. This review briefly summarizes the molecular mechanism of HH signal pathway inducing drug resistance of LSC and leading to novel strategies for eradicating LSC.
Drug Resistance, Neoplasm ; Hedgehog Proteins ; metabolism ; Humans ; Leukemia ; metabolism ; Neoplastic Stem Cells ; drug effects ; Signal Transduction

Hedgehog (Hh) signaling pathway plays an important role during embryonic development and pattern formation. Disruption of Hh pathway results in various developmental disorders and increasing cancer incidence. Here we provide a comprehensive review of the pathway members, focusing on how mammalian Hh regulates the Gli family of transcription factors through its downstream members, the so-called "canonical signaling pathway". Hh signaling pathway is highly conserved among species, and primary cilia plays an important role as a "signaling center" during vertebrate signal transduction. Further, in the past few years, numerous studies have shown that Hh signal can also be transduced through Gli-independent ways collectively referred to as "non-canonical signaling pathways", which can be subdivided into two modules: (i) those not requiring Smo and (ii) those downstream of Smo that do not require Gli transcription factors. Thus, we review the rapid progress on canonical and non-canonical Hh pathways.
Animals ; Cilia ; physiology ; Hedgehog Proteins ; physiology ; Receptors, G-Protein-Coupled ; physiology ; Signal Transduction ; Transcription Factors ; physiology

The Hedgehog (Hh) signalling pathway plays many important roles in development, homeostasis and tumorigenesis. The critical function of Hh signalling in bone formation has been identified in the past two decades. Here, we review the evolutionarily conserved Hh signalling mechanisms with an emphasis on the functions of the Hh signalling pathway in bone development, homeostasis and diseases. In the early stages of embryonic limb development, Sonic Hedgehog (Shh) acts as a major morphogen in patterning the limb buds. Indian Hedgehog (Ihh) has an essential function in endochondral ossification and induces osteoblast differentiation in the perichondrium. Hh signalling is also involved intramembrane ossification. Interactions between Hh and Wnt signalling regulate cartilage development, endochondral bone formation and synovial joint formation. Hh also plays an important role in bone homeostasis, and reducing Hh signalling protects against age-related bone loss. Disruption of Hh signalling regulation leads to multiple bone diseases, such as progressive osseous heteroplasia. Therefore, understanding the signalling mechanisms and functions of Hh signalling in bone development, homeostasis and diseases will provide important insights into bone disease prevention, diagnoses and therapeutics.
Animals ; Bone Development ; Bone Diseases ; metabolism ; Hedgehog Proteins ; metabolism ; Homeostasis ; Humans ; Signal Transduction

Animals ; Hedgehog Proteins ; metabolism ; Humans ; Liver Diseases ; metabolism ; pathology ; Signal Transduction

OBJECTIVES: Silence of SET domain containing lysine methyltransferase 7 (SET7) alleviates myocardial tissue injury caused by ischemia-reperfusion. But the effects of SET7 on angiotensin II (Ang II)-induced myocardial fibroblast proliferation and the collagen synthesis are not clear. The purpose of this study was to explore the effect of SET7 on the proliferation and collagen synthesis of myocardial fibroblasts and its mechanisms. METHODS: Myocardial fibroblasts were isolated and identified by immunofluorescence. Myocardial fibroblasts were randomly divided into 4 groups: a control group (cells were normally cultured), an Ang II group (cells were treated with 100 nmol/L Ang II for 24 h), a siCtrl group (cells were transfected with siRNA control and were then treated with 100 nmol/L Ang II for 24 h), and a siSET7 group (cells were transfected with siRNA SET7 and were then treated with 100 nmol/L Ang II for 24 h). Cell counting kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assay were used to evaluate cell proliferation. Real-time PCR was used to detect the mRNA levels of SET7, collagen I, collagen III, and α-smooth muscle actin (α-SMA). Western blotting was used to detect the protein expression of SET7, collagen I, collagen III, α-SMA, sonic hedgehog (Shh), ptched1 (Ptch1), and glioma-associated oncogene homolog 1 (Gli1). RESULTS: Fluorescence microscopy showed positive vimentin staining, and myocardial fibroblasts were in good condition. As compared to the control group, the mRNA and protein levels of SET7 in the Ang II group were significantly upregulated; cell proliferation rate and EdU fluorescence intensity in the Ang II group were significantly increased; the mRNA and protein levels of collagen I, collagen III, and α-SMA were significantly upregulated (all CONCLUSIONS Silence of SET7 gene inhibits Ang II-induced proliferation and collagen synthesis of myocardial fibroblasts. Shh signaling pathway may be involved in this process.
Angiotensin II/pharmacology* ; Cell Proliferation ; Cells, Cultured ; Collagen/genetics* ; Fibroblasts ; Hedgehog Proteins

OBJECTIVE: To investigate the effects of inhibiting Sonic Hedgehog (Shh) signaling on fibrous scar formation and functional outcome after ischemic brain injury. METHODS: Adult SD rats were randomized into sham-operated group, middle cerebral artery occlusion (MCAO) and reperfusion (I/R) group, I/R with intraventricular empty adenoviral vector (rAd-NC) injection group, and I/R with adenovirus-mediated Shh knockdown (rAd-ShShh) group. After the treatments, the neurological deficits of the rats were assessed, and the protein and mRNA expressions of fibronectin (Fn), α-SMA, and Shh in the ischemic hemisphere were detected with immunofluorescence assay and qPCR; TUNEL staining was used for detecting neural cell apoptosis. In the cell experiment, primary meningeal fibroblasts isolated from neonatal SD rats were pretreated for 24 h with TGF-β1 or TGF-β1 plus cyclopamine (CYC) before oxygen-glucose deprivation for 150 min followed by reoxygenation for 72 h (OGD/R). CCK-8 assay and scratch test were performed to examine the changes in cell proliferation and migration, and immunofluorescence assay, qPCR and Western blotting were used for detecting cell transformation and the expressions of Shh, α-SMA, and Fn. RESULTS: Cerebral I/R injury significantly increased the protein and mRNA expressions of Shh, α-SMA, and Fn in the ischemic hemisphere of the rats, but their expression levels were significantly lowered by intraventricular injection of rAd-Shshh (P < 0.05), which obviously increased cell apoptosis in the ischemic hemisphere (P < 0.05) and improved modified mNSS and modified Bederson scores of the rats (P < 0.05). In the cell experiment, pretreatment with TGF-β1 and TGF-β1+CYC both increased the viability of the primary meningeal fibroblasts after OGD/R. TGF-β1 significantly enhanced the migration ability and induced obvious transformation of the exposed cells (P < 0.05), but these effects were significantly attenuated by co-treatment with CYC (P < 0.05). The expressions of Shh, α-SMA and Fn in the TGF-β1 group were all significantly higher in TGF-β1-treated cells (P < 0.05) and were obviously lowered by co-treatment with CYC (P < 0.05). CONCLUSION Inhibition of Shh signaling may inhibit fibrous scar formation and functional recovery in rats after ischemic brain injury.
Animals ; Brain Injuries ; Cicatrix ; Hedgehog Proteins ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1