Mutations of PTCH1 gene in two pedigrees with bifid rib-basal cell nevus-jaw cyst syndrome / 浙江大学学报·医学版

Two male patients with bifid rib-basal cell nevus-jaw cyst syndrome (BCNS) were admitted to Department of Stomatology, the First Affiliated Hospital of Bengbu Medical College due to radiological findings of multiple low density shadows in the jaw. Clinical and imaging findings showed thoracic malformation, calcification of the tentorium cerebellum and falx cerebrum as well as widening of the orbital distance. Whole exon high-throughput sequencing was performed in two patients and their family members. The heterozygous mutations of c.C2541C>A(p.Y847X) and c.C1501C>T(p.Q501X) in PTCH1 gene were detected in both patients. Diagnosis of BCNS was confirmed. The heterozygous mutations of PTCH1 gene locus were also found in the mothers of the two probands. Proband 1 showed clinical manifestations of low intelligence, and heterozygous mutations of c.C2141T(p.P714L) and c.G3343A(p.V1115I) were detected in FANCD2 gene. Proband 2 had normal intelligence and no FANCD2 mutation. The fenestration decompression and curettage of jaw cyst were performed in both patients. Regular follow-up showed good bone growth at the original lesion, and no recurrence has been observed so far.

Hyalinizing clear cell carcinoma of the soft palate: a review of literature review

Hyalinizing clear cell carcinoma (HCCC), also known as clear cell carcinoma, not otherwise specified [CCC, (NOS)], is a rare minor salivary gland tumor characterized by proliferation of clear cells, organized in trabecular cords, or solid nests within loose to densely hyalinized stroma. It is considered a diagnosis of exclusion by the World Health Organization (WHO) because other salivary tumors may also have a clear cell component. Hence, there is a wide differential diagnosis. EWSR1-ATF1 gene rearrangements are fairly specific for this tumor, however, one of the recent studies have described its presence in clear cell odontogenic carcinoma (CCOC) one of its histologic mimickers. EWSR1 and CREM fusions have recently been described in these tumors but its importance is still not well described. Here we present a case of a 33-year-old woman who presented with a recurrent lesion of the soft palate. Her initial lesion was resected and diagnosed as low-grade myoepithelial tumor. Surgical margins at the time of initial resection were positive and the re-excision was recommended but the patient did not undergo surgery. Two years later, local recurrence at the same site was found and an excision was performed yielding negative margins. Histopathologic examination revealed features consistent with hyalinizing clear cell carcinoma. The patient remains disease free 1 year after the re-excision. The pathology, clinical characteristics, differential diagnosis and treatment of hyalinizing clear cell carcinoma are reviewed.

Unveiling The Role Of Shh From Embryogenesis To Its Reactivation In Tumorigenesis:A Maxillofacial Perspective.

Sonic Hedgehog gene (SHH) plays a vital role in embryogenesis through its secreted protein sonic hedgehog protein (Shh). During embryogenesis, Shh acts as a morphogen controlling proximal and distant signaling in the specific development of tissue lineages, patterning, regulation of cell proliferation and suppression of tissue apoptosis. Shh also exerts its role in odontogenesis by determining the site of tooth bud formation, in tooth morphogenesis and root formation. The difference in the specific development of a region by Shh can be explained by its [a] 'Spatial gradient [b] the 'form' [c] Concentration gradient and [d] Temporal gradient. Shh signaling pathway has an extracellular and an intracellular component. A disruption of Shh pathway contributes to tumorigenesis of several cell types including those arising from odontogenic structures. This article reviews Shh from its formation in embryonic stages, its role in development and odontogenesis, to its reactivation in tumorigenesis and in specific to odontogenic pathologies.

Mechanism of polypeptide extract from scorpion venom inhibiting Hedgehog signaling transduction in K562/BALB/c-nu leukemia mice / 中草药

Objective To study the mechanism of the Hedgehog signaling transduction intervened by polypeptide extract from scorpion venom (PESV) on K562/BALB/c-nu leukemia mice. Trying to analyze the molecular mechanisms and targets of the inhibited effect of PESV on chronic myeloid leukemia (CML) in vivo. Methods After establishing the K562/BALB/c-nu leukemia mice successfully, the model mice were divided into six groups which were the blank group, the PESV high, medium, and low doses (0.3, 0.6, 1.2 mg/kg) group, the Imatinib (50 mg/kg) group, and the model group. After 14 d drug intervention, the levels of gene and protein expression of Hedgehog signaling pathway upstream factors Shh, Ptch, and Smo were detected by qRT-PCR and Western blotting, and the protein expression of downstream factor Gli1 was determined by ELISA test. Results Compared to the model group, the genetic and protein expression of Shh which was an upstream factor were increased in the PESV groups. The mRNA and protein expression of Ptch and Smo in PESV low-dose and medium-dose groups were decreased. There were no significant differences of upstream factors between Imatinib group and model group. The concentration of downstream Gli1 protein significantly decreased within low-dose and medium-dose PESV groups, while there was no significant difference between high-dose PESV group and Imatinib group. Conclusion PESV can inhibit the expression of Hedgehog signaling pathway upstream factor Ptch, Smo and downstream factor Gli1 on the mRNA and protein level, while Imatinib has no obvious inhibitory effect on the Hedgehog signaling pathway.

Identification of PTCH1-3'-UTR regulated lncRNAs associated with competing endogenous RNA networks in non-small cell lung cancer cell line / 复旦学报（医学版）

Objective To investgate the effect of PTCH1-3'-UTR on the expression of long noncoding RNAs (lncRNAs) and analyze regulatory networks so as to indicate the function of PTCH1-3'-UTR.Methods We screened PTCH1-3'-UTR regulated lncRNAs in non-small cell lung cancer (NSCLC) by using microarray,and validated the expression by qPCR.To explore the potential mechanisms of these lncRNAs underlying NSCLC progression,we performed GO and KEGG pathway analysis of the dysregulated lncRNAs.We also conducted a bioinformatic analysis in TCGA database to identify the association of PTCH1-3'-UTR regulated lncRNAs and the overall survival of NSCLC patients.Results The expression of seven PTCH1-3'-UTR up-regulated lncRNAs (LOC100507547,FAM41C,DOCK4-AS1,AC009305.1,KLF7-IT1,RP11-749H20.1,LINC01511) were validated by qPCR.The GO and KEGG pathway analysis of the dysregulated lncRNAs indicated that a series of biological progresses were involved in the function of PTCH1-3'-UTR regulated lncRNAs,including transcription,signal transduction,protein transport and translational elongation,and several pathways,such as calcium signaling pathway,Jak-STAT signaling pathway,p53 signaling pathway and insulin signaling pathway.Among the lncRNAs regulating PTCH1-3'-UTR,6 were shown to be associated with the overall survival of NSCLC patients.High expression of lncRNA-CDKN2BAS and FAM66D related to the probability of lower survival,while high expression of lncRNA-LINC00240,LOC400027,ABCC6P2 and FLJ10038 might have a higher probability of survival.Conclusions The study would provide an insight of the function of PTCH1-3'-UTR,and PTCH1-3'-UTR dysregulated lncRNAs may be potential prognostic biomarkers for NSCLC.

Novel Patched 1 Mutations in Patients with Gorlin-Goltz Syndrome Strategic Treated by Smoothened Inhibitor

We studied a family with Gorlin-Goltz syndrome. The novel mutations of our cases were located on the 21st exon of the PTCH1 gene (c.3450C>G). The father, who received a strategic 56-day vismodegib treatment for disease control, was the first patient with Gorlin syndrome treated with the hedgehog inhibitor in Taiwan. The lesions regressed gradually, with scar formation, and were subsequently removed via a wide excision. Further details are provided below.

Clinical characteristics and genetic analysis of nevoid basal cell carcinoma syndrome with epilepsy / 上海交通大学学报（医学版）

Objective • To investigate the clinical and genetic characteristics of nevoid basal cell carcinoma syndrome (NBCCS) combined with epilepsy. Methods • The clinical data of a proband with the symptom of epileptic seizures in Department of Neurology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine was retrospectively analyzed. Facial nevoid was resected and hematoxylin-eosin (H-E) staining was examined. Wholeexome sequencing was performed on the blood DNA of the proband and his family members. Sanger sequencing was used for co-segregation analysis. Results • The proband was highly suspected of having NBCCS according to the clinical diagnostic criteria of NBCCS. H-E staining showed typical pathological features of basal cell carcinoma. Heterozygous deletion at c.3364_3365del base in the exon 20 of patched 1 (PTCH1) gene was found in this family. Conclusion • The heterozygous deletion in the exon 20 of PTCH1 gene (c.3364_3365del) may be a potential hot spot mutation for NBCCS, especially in patients combined with neurological manifestations, such as epilepsy.

Effect of Mono-2-ethyhexyl Phthalate on DNA Methylation in Human Prostate Cancer LNCaP Cells / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate whether mono (2-ethylhexyl) phthalate (MEHP) affects genomic DNA methylation and the methylation status of some specific genes such as patched gene (PTCH) and smoothened gene (SMO) in LNCaP cells.</p><p><b>METHODS</b>LNCaP cells were treated with MEHP (0, 1, 5, 10, and 25 μmol/L) for 3 days. An ELISA assay was preformed to detect genomic methylation, including 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) content. A pyrosequencing assay was applied to assess DNA methylation in PTCH and SMO gene promoters. The correlation between DNA methylation and gene expression was assessed.</p><p><b>RESULTS</b>The proportion of cytosines with 5-mC methylation in LNCaP cells was significantly decreased by MEHP (1, 5, 10, and 25 μmol/L) in a dose-dependent manner (P < 0.01). For genes in the Hedgehog pathway, there was no significant MEHP concentration-dependent difference in the DNA methylation of PTCH and SMO.</p><p><b>CONCLUSION</b>MEHP might affect the progression of prostate cancer through its effect on global DNA methylation.</p>

Effects of PTCH1 mutations on the epithelial proliferation derived from keratocystic odontogenic tumour / 北京大学学报(医学版)

Objective: To explore the relationship between the PTCH1 mutation and the expression of bcl-2, filaggrin, and loricrin in the keratocystic odontogenic tumour (KCOT), as well as the effects of the mutated PTCH1 on the epithelial proliferation and differentiation.Methods: The samples were collected from 20 cases of KCOT with mutated PTCH1, as well as 20 cases without mutation.All the samples were analyzed with immunohistochemical staining, for the purpose of investigating the expression of bcl-2, filaggrin, and loricrin.Results: In the samples with mutated PTCH1, the epithelia of 60% (12/20) cases expressed intensively positive bcl-2 staining, 20% (4/20) expressed moderate staining, and 20% (4/20) weak staining, but no negative bcl-2 staining samples were investigated;it was significantly different from the samples without PTCH1 mutation, in which 20% (4/20) expressed intensive staining, no moderate staining, 50% (10/20) weak staining, and 30% (6/20) negative staining were investigated (U=72, P=0.001).For the expression of filaggrin, 55% (11/20) of samples with PTCH1 mutations were stained weakly and 45% (9/20) showed negative staining, while in the samples not harboring PTCH1 mutations, 30% (6/20) cases showed moderate positive staining, 40% (8/20) weak staining and 30% (6/20) negative staining, no intensive staining was investigated (U=182, P=0.48).The loricrin expressed in all the layer of the epithelia in all the samples, while the filaggrin was mainly loca-lized within 1-4 layer cells of the epithelia.The differences of the expression of filaggrin and loricrin between the samples with mutated PTCH1 and without mutated PTCH1 displayed no significance.Conclusion: In the epithelia of KCOT, the bcl-2 expression was significantly associated with the PTCH1 mutation, which suggested that the mutated PTCH1 gene perhaps promotes the proliferation of KCOT epithelium.

Patched-1 and Smoothened, a Hedgehog Receptor and Signal Transducer are Highly Expressed in Diffuse Large B-Cell Lymphoma

Introduction: The Hedgehog (Hh) signalling pathway is a developmental signalling pathway involved in normal mammalian developmental and homeostasis of adult renewable tissues. In most adult tissues, this pathway remains silent and previous studies have shown that constitutive activation of Hedgehog signalling pathway leads to various types of malignancies including medulloblastomas, basal cell carcinoma, gastrointestinal, breast and prostate cancer. The purpose of this study was to investigate the immunohistochemical expression of Hedgehog pathway proteins in Diffuse Large B-cell Lymphoma and determine their association with overall survival (OS). Methods: Positive control using normal tonsils were included in each batch of immunohistochemical staining procedure. Results: PTCH1 proteins were highly expressed in DLBCL and showed strong staining intensity in 107 (100%) cases and SMO proteins were expressed in 105 (98.1%) cases. PTCH1 proteins were localised in the nucleus of tumour cells, whereas SMO proteins were mainly localised in the cytoplasm of tumour cells. Positive expression of PTCH1 and SMO proteins and overall survival of DLBCL patients were correlated with age, gender, race and tumour location. There was no significant correlation between the expression of these two proteins with any of the parameters. PTCH1 expression showed significant association with SMO expression (P=0.03). Conclusions: Our findings suggest that high expression of both PTCH1 and SMO may be important in the pathogenesis of DLBCL. However, additional mechanisms that may contribute to the activation of HH signalling in DLBCL needs to be further explored.

A novel PTCH1 gene mutation in a pediatric patient associated multiple keratocystic odontogenic tumors of the jaws and Gorlin–Goltz syndrome.

Gorlin–Goltz syndrome (GGS) is an uncommon autosomal dominant inherited disorder which comprises the triad of basal cell carcinomas (BCCs), odontogenic keratocysts, and musculoskeletal malformations. Besides this triad, neurological, ophthalmic, endocrine, and genital manifestations are known to be variable. It is occasionally associated with aggressive BCC and internal malignancies. This report documents a case of GGS with a novel mutation in the PTCH1 gene in an 11‑year‑old child. The clinical, radiographic, histopathologic and molecular findings of this condition, and treatment are described, and a review of GGS was carried out.

Síndrome de carcinoma basocelular nevoide con agenesia de cuerpo calloso, mutación en PTCH1 y ausencia de carcinoma basocelular

El síndrome del carcinoma basocelular nevoide (SCBCN) o de Gorlin-Goltz es un raro desorden autosómico dominante con un amplio espectro de manifestaciones clínicas. El signo cardinal es la presencia de múltiples carcinomas basocelulares (CBCs) y su ausencia demora el diagnóstico. Presentamos un adolescente de 14 años con diagnóstico de SCBCN por la presencia de queratoquistes odontogénicos, hiper­telorismo, macrocefalia y agenesia del cuerpo calloso pero sin lesiones cutáneas. La madre, de 43 años, tiene diagnóstico de SCBCN y no presenta CBCs. Para completar el estudio se realizó secuenciación bidireccional y Multiplex Ligation dependent Probe Amplification (MLPA) en sangre periférica para buscar mutaciones en PTCH1, principal gen responsable del síndrome. Se encontró una mutación germinal novel en el paciente y la madre: una duplicación de 25 pb en el exón 10 (c.1375dupl25bp). El análisis bioinformático predijo un corrimiento del marco de lectura y un codón stop prematuro, que produciría una proteína trunca más corta que lo normal. Nuestros resultados sugieren que el estudio clínico y genealógico completo con análisis genético es fundamental para la detección temprana de casos como el presente.

Nevoid Basal Cell Carcinoma Syndrome (NBCCS) or Gorlin-Goltz syndrome is a rare autosomal dominant disorder, mainly due to PTCH1 gene mutations, that comprises a broad spectrum of clinical manifestations. The presence of multiple basal cell carcinomas (BCCs) is a cardinal sign in NBCCS, therefore cases in which BCCs are absent entails a delay in the diagnosis.We present a 14 years old boy with a clinical diagnosis of NBCCS by the presence of odontogenic cysts, hypertelorism, macrocephaly, and corpus callosum agenesia, but with absence of skin lesions. His 43 years old mother has NBCCS diagnosis and no history of BCCs. For a deeper study, PTCH1 mutation screening from peripheral blood samples were performed by both bidirectional sequencing and multiplex ligation dependent probe amplification (MLPA) techniques. The proband and his mother carry 25 pb duplication in exon 10 (c.1375dupl25bp) that causes a reading frameshift with a premature stop codon. Bioinformatics analysis predicted that this mutation results in a truncated protein shorter than normal. Our results suggest that complete clinical and genealogical studies accompanied by genetic analysis are essential in the early detection of the NBCCS cases such the one presented here.

Mutation analysis of the PTCH1 gene in a pedigree with nevoid basal cell carcinoma syndrome / 中华皮肤科杂志

Objective To analyze mutations in the PTCH1 gene in a pedigree with nevoid basal cell carcinoma syndrome (NBCCS).Methods Blood samples were collected from a 58-year-old male proband with NBCCS (Ⅱ 5),his brothers (Ⅱ 1 and Ⅱ 3) and son (Ⅲ4),and 50 unrelated healthy human controls.DNA was extracted from these blood samples.PCR and direct DNA sequencing were performed to determine mutation sites in the PTCH1 gene.According to the mutation sites,allele-specific oligonucleotide primers were designed and used to confirm the pathogenic mutations in this pedigree through PCR.Results A nonsense mutation (c.2137C),which leads to the substitution of CAG by TAG with the generation of a premature termination codon (Q714X),was identified in exon 14 in one allele of the PTCH1 gene in the proband and his son,but in none of the healthy human controls.Conclusion The nonsense mutation (c.2137C ＞ T) in the PTCH1 gene may be a specific mutation causing the clinical symptoms in the patient with NBCCS.

Immunohistochemical analysis of human homologue of drosophila patched (PTCH) in dental follicles of impacted third molars / Análisis imunohistoquímico del homólogo humano del gen parcheado de drosofila (PTCH) en folículos dentales de terceros molares retenidos

This study investigated the immunodetection of PTCH in epithelial components of dental follicles associated with impacted third molars without radiographic signs of pathosis. One hundred and five specimens of dental follicles associated with impacted third molars with incomplete rhizogenesis (between Nolla's stage 6 and 9) were surgically removed from 56 patients. Epithelial cell proliferation was determined by using immunohistochemical labeling. Statistical analysis was performed using Fisher exact test and a level of significance of 5 percent. Of the 105 dental follicles collected, 3 were PTCH-positive. The specimens with squamous metaplasia and epithelial hyperplasia had higher rates of positivity for PTCH, as well as those with active remnants of odontogenic epithelium. This study suggests that the odontogenic cells of the dental follicle might be proliferating during the rhizogenesis, while the squamous metaplasia and hyperplasia of the epithelial lining and proliferative odontogenic epithelial rests show the differentiation potential of dental follicles.

Se investigó la inmunodetección de PTCH en los componentes epiteliales de los folículos dentales asociados a terceros molares retenidos sin signos radiográficos y morfológicos de patología. Fueron quirúrgicamente extraídos de 56 pacientes 105 muestras de folículos dentales asociadas a terceros molares retenidos con rizogénesis incompleta (entre el estadio de Nolla 6 y 9). La proliferación de células epiteliales se deteminó mediante inmunohistoquímica. El análisis estadístico se realizó mediante la prueba exacta de Fisher. De los 105 folículos dentales recogidos, 3 fueron PTCH-positivos. Las muestras con metaplasia escamosa e hiperplasia epitelial tuvieron mayores tasas de positividad para PTCH, así como aquellos con los restos de proliferación del epitélio odontogénico. En conclusión, este estudio sugiere que las células odontogénicas del folículo dental podrían estar proliferando durante la rizogénesis, mientras que la metaplasia escamosa e hiperplasia del epitelio y de restos epiteliales odontogénicos en proliferación muestran el potencial de diferenciación de los folículos dentales.

Expression of GLI1 and PTCH1 mRNA and yts correlation with clinical parameters in pancreatic cancer / 中华胰腺病杂志

Objective To investigate the expression of GLI1 and PTCH1 mRNA in pancreatic cancer and study its clinical significance. Methods Real-time fluorescence quantitative PCR (RFQ-PCR) was used to detect the expression of GLI1 and PTCH1 mRNA in 35 samples of pancreatic cancer tissues and 27 samples of adjacent normal pancreatic tissues, and the correlation of GLI1 and PTCH1 mRNA expression with clinical parameters was investigated. Results The relative expression of GLI1 mRNA in pancreatic cancer tissues was 1.12 ～ 3. 65 ( median 1.19), the relative expression of TCH1 mRNA was 1.82 ～ 4.36 ( median 2.36 ). The relative expression of GLI1 mRNA in adjacent normal pancreatic tissues was 0.23 ～ 2.76 ( median 0.87 ), the relative expression of PTCH1 mRNA was 1.11 ～ 2. 17 (median 0.58). Both the expression of GLI1 and PTCH1 mRNA in pancreatic cancer tissues were significantly higher than those in normal pancreatic tissues (P<0.05), and a positive correlation was found between GLIl and PTCH1 mRNA expression levels (P <0.05 ). The expression of GLI1 mRNA was significantly correlated with the differentiation degree and lymph node metastasis of pancreatic cancer (P < 0. 05). Conclusions GLI1 and PTCH1 may be involved in pancreatic carcinogenesis, and GLI1 may be related to invasion and lymph node metastasis of pancreatic cancer.

Role of PTCH1 gene hypermethylation in tumorigenesis of gastric cancer / 第二军医大学学报

Objective: To study the effect of PTCH1 methylation on gastric carcinogenesis and the therapeutic effect of methylation inhibitor, 5-aza-2′-deoxyeytidine (5-aza-dC), for treatment of gastric cancer. Methods: The total RNAs were extracted from 10 gastric cancer tissues, their corresponding adjacent normal tissues, and gastric cancer cell line AGS. The PTCH1 mRNA expression was detected by Quantitative real-time PCR (QRT-PCR) and the methylation of the promoter was examined by methylation specific PCR (MSP). AGS cells were treated by 5-Aza-dC; the cell cycle and apoptosis were examined by flow cytometry, and the methylation level was also observed. Results: PTCH1 expression was negatively correlated with promoter methylation in gastric cancer tissues, their corresponding adjacent normal tissues, and gastric cancer cell line AGS (r = -0.591, P = 0.006). 5-Aza-dC treatment caused apoptosis and G 0/G1 phase arrest of AGS cells, and also induced demethylation of PTCH1 and increased its expression. Conclusion: Hypermethylation of PTCH1 gene promoter region is one of the main causes of low PTCH1 expression in AGS cells. Demethylation agent 5-Aza-dC can reverse this methylation status of PTCH1 and regulate the expression of PTCH1, suggesting a role for it in gastric cancer treatment.

Expression of Shh and Ptch gene in gallbladder carcinoma / 国际外科学杂志

Objective Shh and Ptch have been known to play critical roles in the hedgehog pathway and the hedgehog pathway activation occurs in the gastrointestinal cancers. This study was to detect the expression of Shh and Ptch in gallbladder carcinoma, and explore their correlation to gallbladder carcinoma. Methods The expression of Shh and Patch protein in 41 specimens of primary gallbladder carcinoma, 20 specimens of normal gallbladder and 21 specimens of adenoma tissue were assessed by immunohistochemistry.Results The positive expression rate of Shh and Ptch in gallbladder carcinoma was 75.6% (31/41) and 78.0% (32/40) respectively, in normal gallbladder mucous was 5% ( 1/20 ) and 5% ( 1/20 ) respectively,in gallbladder adenoma was 4.7% ( 1/21 ) and 9.6% (2/21) respectively, and there was significant differenee between the three groups in Shh and Patch expression values(P ＜0.001 ). However, there was no significant difference between age group, histological grade group, histologic type group, Nevin stage group,lymph node or distal organ metastasis group, and gallstone presence group in Shh and Ptch expression value (P ＞ 0.05, respectively). The expression of Shh was positively correlated to Ptch (r = 0.72, P ＜ 0.01 ).Conclusion These data support our hypothesis that Hh signaling is dysregulated in human gallbladder carcinognesis.

Basal cell nevus syndrome (gorlin syndrome) confirmed by PTCH mutations and deletions / 소아과

Basal cell nevus syndrome (BCNS), also known as Gorlin syndrome, is a rare autosomal dominant disorder characterized by variable manifestations, including multiple basal cell carcinomas, odontogenic keratocysts of the jaw, skeletal anomalies including scoliosis and bifid ribs, palmar and plantar pits, calcification of the falx cerebri, and biparietal frontal bossing. We report a case of a 9-year-old boy with the clinical features of basal cell nevus syndrome, in which a PTCH gene mutation was confirmed by DNA testing.