Mutación de protocadherina 19 (PC-DH19), en paciente con epilepsia refractaria / Mutation of protocadherin 19 (PCDH19), in patients with refractory epilepsy

Paciente de 4 años de edad, con epilepsia de difícil manejo, cuya etiología se atribuye a patología autoinmune y que finalmente se diagnostica una mutación de protocadherina (PCDH19). Se discute la fisiopatología, características clínicas, exámenes y los posibles tratamientos.

Four-year-old patient with intractable epilepsy, whose etiology is attributed to autoimmune pathology and who is eventually diagnosed with a protocadherin mutation (PCDH19). Pathophysiology, clinical characteristics, examinations and possible treatments are discussed.

Effect of protocadherins 10 re-expression induced by 5-aza-2'-deoxycytidine on invasion and migration capacity of breast cancer cell line MDA-MB-231 and its mechanism / 解剖学报

Objective: To investigate whether re-expression of protocadherin 10(PCDH10) induced by 5-aza-2'-deoxycytidine (5-Aza-CdR) could affect the invasion and migration of MDA-MB-231 cells, and to explore the possible mechanism. Methods: Human breast cancer cell line MDA-MB-231 was cultured in vitro. Control group and 5-Aza-CdR treatment group were set up. PCDH10 mRNA expression in MDA-MB-231 cell line was determined by reverse transcriptionpolymerase chain reaction (RT-PCR); Transwell chamber and wound healing assay were performed to measure the invasion and migration capacity of the cells, and protein expression of PCDH10, DNA methyltransferase(DNMT)3A, DNMT3B, nuclear factor(NF)-κB p65, matrix metalloproteinases(MMP)-2 and MMP-9 were detec Western blotting. Results: 5- Aza-CdR could reverse the methylation status of PCDH10 gene in MDA-MB-231 cells in a dose-dependent manner. Reexpression of PCDH10 significantly inhibited cell invasion and migration capacity in vitro. Western blottoing analysis revealed that the expression of DNMT3A, DNMT3B, NF-κB p65, MMP-2 and MMP-9 in MDA-MB-231 cells were downregulated after exposure to 5-Aza-CdR. Conclusion: Re-expression of PCDH10 significantly inhibits MDA-MB-231 invasion and migration capacity. The inhibitory effect is characterized that 5-Aza-CdR treatment down-regulates DNMT3A and DNMT3B levels, recovers the expression of anti-oncogene PCDH10, further blocks the activation of NF-κB p65, resultsing in a decrease in the secretion of MMP-2 and MMP-9.

Tichostatin A inhibiting migration and invasion of human gastric carcinoma SGC-7901 cells via up-regulating protocadherin 9 / 解剖学报

Objective To investigate the effect of trichostatin A (TSA) on migration and invasion in human gastric carcinoma SGC-7901 cells and its possible mechanism. Methods SGC-7901 cells were cultured in vitro and treated with TSA (5, 10, 20, 40, 80, 160 nmol/L) for 48 hours, and then the cell viability was detected by cell counting kit 8 (CCK-8) assay. The protocadherin 9 (PCDH9) high-expression SGC-7901 cells were stably established by transfecting with eukaryotic expression vector (pCMV6-PCDH9). Transwell assay was used to determine the abilities of migration and invasion. The mRNA expression level of PCDH9 were measured by RT-PCR. Western blotting was performed to analyze the protein expression of PCDH9, Snail, E-cadherin, matrix metalloproteinase (MMP)-2 and MMP-9. Results TSA remarkably reduced the cell viability of SGC-7901 cells in excess of 80 nmol/L (P<0. 05). However, in a dose-dependent manner, low-level TSA (5-20 nmol/L) suppressed migration and invasion of SGC-7901 cells (P<0. 05), down-regulated the protein levels of Snail, MMP-2 and MMP-9 (P<0. 05), and up-regulated the protein levels of PCDH9 and E-cadherin (P<0. 05). Meanwhile, high expression of PCDH9 also inhibited migration and invasion of SGC-7901 cells (P<0. 05), down-regulated the protein levels of Snail, MMP-2 and MMP-9 (P<0. 05), and up-regulated the protein level of E-cadherin (P<0. 05). Conclusion TSA may inhibit migration and invasion of SGC-7901 cells most likely via up-regulating PCDH9, and then down-regulating the protein levels of Snail, MMP-2 and MMP-9, and up-regulating the protein level of E-cadherin.

Expression of protocadherin 10 in non-small cell lung cancer and effect of protocadherin 10 over-expression on function of A549 cells / 肿瘤

Objective: To investigate the expression of protocadherin 10 (PCDH10) in non-small cell lung cancer (NSCLC), and to explore the function and molecular mechanism of PCDH10 in NSCLC cells. Methods: The expression levels of PCDH10 and Ki67 mRNAs in NSCLC tissues and paracancerous normal tissues were detected by real-time fluorescent quantitative PCR. The correlations of PCDH10 and Ki67 mRNA expressions with the clinicopathological features and prognosis of NSCLC patients were analyzed. A549 cells were transfected with recombinant plasmid pcDNA3.1(+)-PCDH10 or empty vector pcDNA3.1(+) (as the control) by LipofectAMINE 2000. The proliferation, apoptosis and cell cycle were detected by IncuCyte S3 live-cell analysis system and FCM method, respectively. The migration and invasion abilities were detected by Transwell method. The expression levels of proliferating cell nuclear antigen (PCNA), Bcl-2, cyclin D1, cyclin E, cyclin-dependent kinase 4 (CDK4), E-cadherin and Slug mRNAs and proteins were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. Results: The expression level of PCDH10 mRNA in NSCLC tissues was lower than that of para-cancerous normal tissues (P < 0.001). The expression level of PCDH10 mRNA in NSCLC tissues was negatively correlated with T stage, lymph node metastasis, clinical stage and Ki67 mRNA expression (all P < 0.001), and positively correlated with prognosis of NSCLC patients (P < 0.001). PCDH10 over-expression inhibited the proliferation of A549 cells by blocking cell cycle at G1 phase and promoting apoptosis (all P < 0.05). PCDH10 significantly inhibited the invasion and migration of A549 cells (both P < 0.05). The expression levels of PCNA, cyclin D1, cyclin E, CDK4, Bcl-2 and Slug mRNAs and proteins were downregulated in A549 cells with PCDH10 over-expression (all P < 0.05), but the expression levels of E-cadherin mRNA and protein were up-regulated (both P < 0.05). Conclusion: PCDH10 is lowly expressed NSCLC tissues. PCDH10 over-expression can inhibit the proliferation and invasion of NSCLC cells.

Progress of protocadherin 19(PCDH19)gene related epilepsy / 中华实用儿科临床杂志

PCDH19 gene related epilepsy is an unusual X - linked disease that females and mosaic males are affected,while hemizygous males are not. Recently,the number of reports about PCDH19 gene related epilepsy is in-creasing,and PCDH19 gene has become one of the most important epilepsy genes. Now,the structure and function of PCDH19 gene and protein,the inheritance and pathogenesis,clinical features,treatment and genotype/ phenotype asso-ciated with PCDH19 gene related epilepsy,were reviewed.

PCDH19 gene mutations lead to epilepsy with mental retardation limited to females in 2 cases and literature review / 中南大学学报(医学版)

Epilepsy with mental retardation limited to females (EFMR) is a syndrome characterized by early onset heat-sensitive epilepsy of infancy or early childhood and generally limited to females,which previously reported that the cadherin gene superfamily subtype protocadherin 19 (PCDH19)gene is its pathogenic gene.We retrospectively analyzed the clinical data for 2 cases of EFMR patients with PCDH19 mutation diagnosed by Department of Pediatric Neurology of Xiangya Hospital,Central South University in 2015.Literature on PubMed,OMIM and HGMD relevant to this syndrome was reviewed,and the clinical characteristics were summarized accordingly.The 2 cases are consistent with the typical clinical manifestations of EFMR caused by PCDH19 mutations.Their seizures are heat sensitive,with or without screaming,and expressed in various forms.Cognitive impairment or autism-like performance were often identified in these patients,hematuria metabolic diseases screening was normal,no abnormal MRI imaging of the head,and de novo PCDH19 gene mutations were found in their epilepsy gene chip sequencing.It is noteworthy that this disease is very similar to the clinical manifestations of the Dravet syndrome due to the mutations of the neurotype sodium channel αl subunit SCN1A.Therefore,in female patients whose clinical manifestations resemble to Dravet syndrome but SCN1A gene test were negative,EFMR with PCDH19 mutation should be taken into consideration.Early PCDH19 gene testingis of great significance because it not only helps clinicians to understand and analyze the prognosis of this disease,but also offers genetic counseling to the parents.

Correlation between mRNA expression of protocadherin-10 and prognosis in gastric cancer / 中国癌症杂志

Background and purpose:Promoter methylation ofPCDH10, a gene encoding protocadherin 10, has been found to be correlated to poor prognosis in gastric cancer (GC) patients. However, the relationship between the expression of PCDH10 and prognosis in GC remained unknown. This study aimed to explore the relationship be-tween the expression of PCDH10 and clinicopathological features and prognosis of GC, and to identify biomarker for predictions of recurrence and survival of GC.Methods:mRNA expressions of PCDH10 in 115 pairs of GC tissues and adjacent normal tissues were detected by real-time lfuorescence quantitative polymerase chain reaction (RTFQ-PCR). The correlation between PCDH10 expression level and clinicopathological features and prognosis of GC was analyzed. Prediction models for 5-year recurrence and 5-year survival were established using logistic regression method.Results:Progression-free survival (PFS) and overall survival (OS) were signiifcantly prolonged in patients with PCDH10 low expression compared to patients without PCDH10 low expression (P=0.046 andP=0.033 respectively). PCDH10 low expression signiifcantly correlated with less lymph node metastasis (P=0.001) and earlier TNM staging (P=0.001), and was more common in female than in male (P=0.040). The mRNA expression of PCDH10 did not correlate with age, Lauren classiifcation, T stage, neural invasion or vascular invasion. Univariate Cox analysis showed Lauren classiifca-tion, T stage, N stage, M stage and PCDH10 expression signiifcantly correlated with PFS and OS. Logistic regression models for the prediction of 5-year recurrence or 5-year survival based on clinicopathological features included Lauren classiifcation, T stage, N stage and M stage as variables. Logistic regression models for the prediction of 5-year recur-rence or 5-year survival based on PCDH10 expression included Lauren classiifcation, T stage, M stage and PCDH10 expression level but not N stage as variables. The models based on PCDH10 expression had the same effciencies as models based on clinical parameters in predicting 5-year recurrence or 5-year survival for GC patients.Conclusion:PCDH10 low expression correlated with better prognosis, less lymph node metastasis and earlier TNM stage in GC patients. Low expression of PCDH10 may be a biomarker of better survival for GC patients. Logistic regression model based on PCDH10 mRNA expression may serve as a prediction model when patients have unknown lymph node metas-tasis status.

Methylation of PCDH8 in pancreatic carcinoma cell lines / 中华胰腺病杂志

Objective To investigate the methylation status of PCDH8 gene in pancreatic carcinoma.Methods Methylation of PCDH8 gene in 2 samples of normal pancreatic tissues and 6 pancreatic carcinoma cell lines (PANC1, ASPC1, BxPC3, CFPAC, PaTu8988 and SW1990) was detected by the methylationspecific PCR (MSP) method. The expression of PCDH8 mRNA was detected with 5-Aza-2-deoxycytidine (5-Aza-dC) treatment, a kind of DNA methyltransferase (DNMT) inhibitor in 6 pancreatic carcinoma cell lines by real-time-PCR. Results The methylation of PCDH8 gene was not detected in normal tissues, while it was partially methylated in PANC1, BxPC3, CFPAC and it was totally methylated in PaTu8988, ASPC1, SW1990.PCDH8 mRNA was expressed in PANC1, SW1990, PaTu8988 and the relative quantities of mRNA expression (RQ) were 1.576 ± 0.648, 0.013 ± 0.008, 0.002 ± 0.001; PCDH8 mRNA was not expressed in BxPC3,CFPAC, ASPC1. After 5-Aza-dC treatment, PCDH8 mRNA was expressed in PANC1, ASPC1, BxPC3,CFPAC, PaTu8988, SW1990 and the relative quantities of mRNA expression all significantly increased, and they were 7. 463 ± 2.628, 10. 696 ± 1.539, 7.852 ± 2.762,421.815 ± 1.493, 118.595 ± 4.089, 6.690 ±1.884. Conclusions The methylation of PCDH8 gene may be the major mechanism of down-regulated expression of PCDH8 gene in pancreatic carcinoma.

Preparation and Characterization of Polyclonal Antibody Against Xenopus PAPC / 生物化学与生物物理进展

Xenopus Paraxial Protocadherin (PAPC), which was initially identified in a screen for genes present in the Spemann organizer of Xenopus embryos, is required for gastrulation, somitogenesis and otic vesicle formation. In order to investigate its function in various developmental events, an antibody was prepared which could specifically recognize Xenopus PAPC. Glutathione S transferase (GST) expression system was used to express the fusion protein GST-PAPC. Rabbits were immunized with GST-PAPC Western blotting analysis of FL-PAPC transfected HEK 293T cells lysates, which could be specifically blocked by pre-adsorption of prokaryotic expressed GST-PAPC fusion protein. Furthermore, by using immunofluorescence analysis the polyclonal antibody recognized membrane-bound PAPC in FL-PAPC transfected 293T cells and Xenopus animal cap cells. By Western blotting analysis,the endogenous 150 ku PAPC protein was detected in Xenopus embryos using the anti-PAPC antibody. Take together it could be concluded that a polyclonal antibody specifically against Xenopus PAPC was developed.