Analysis of ZEB2 gene variation in two patients with Mowat-Wilson syndrome / 中华医学遗传学杂志

OBJECTIVE@#To identify pathogenic variants in two patients with suspected for Mowat-Wilson syndrome (MWS).@*METHODS@#Genomic DNA was extracted from peripheral blood samples of the patients and his family members, and gene variants were analysis by Trio-whole exome sequences and copy number variation sequencing.@*RESULTS@#Patient 1 was found to carried a de novo heterozygous c.2769C>A (p.Y923*) nonsense variant of ZEB2 gene. The variant was not found in his healthy parents and sister. Patient 2 carried a de novo heterozygous frameshift variant of the ZEB2 gene, namely c.315delC (p.A105Afs*3), which has not been previously reported. Both variants were predicted to be pathogenic and can lead to premature occurrence of stop codons.@*CONCLUSION@#The heterozygous c.2769C>A (p.Y923*) and c.315delC (p.A105Afs*3) variants of the ZEB2 gene probably underlay the pathogenesis in the two patients. Gene testing has facilitated confirmation of the diagnosis and genetic counselling.

Analysis of a case with Mowat-Wilson syndrome caused by ZEB2 gene variant / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis of a proband with distinctive facial features, global developmental delay, seizures and hypoplasia of corpus callosum through next generation sequencing (NGS).@*METHODS@#Genomic DNA was extracted from peripheral blood samples of the proband and his family members. Whole exome and flanking sequences were screened by NGS. Suspected variants were verified by Sanger sequencing.@*RESULTS@#The proband was found to carry a heterozygous c.2824G>T (p.G942X) variant of the ZEB2 gene, which was verified by Sanger sequencing to be a de novo variant.@*CONCLUSION@#The heterozygous c.2824G>T (p.G942X) variant of the ZEB2 gene probably underlies the Mowat-Wilson syndrome in the proband.

Prenatal diagnosis of a fetus with Mowat-Wilson syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a fetus featuring increased nuchal thickness.@*METHODS@#Routine G-banding karyotyping and single nucleotide polymrophism array were carried out to detect genomic copy number variations (CNVs) in the fetus.@*RESULTS@#The fetus was found to harbor a heterozygous 3.8 Mb deletion in the 2q22.2-q22.3 region encompassing the ZEB2 gene, which is closely associated with Mowat-Wilson syndrome (MWS).@*CONCLUSION@#Haploinsufficiency of the ZEB2 gene may predispose to MWS. Lack of knowledge regarding to the ultrasonographic features of MWS may lead to misdiagnosis of the syndrome.

Effect of miR-145 on Migration and Invasion of Ovarian Cancer Cells / 中国医学科学院学报

Objective To evaluate the effect of miR-145 on migration and invasion of ovarian cancer cells.Methods The effect of miR-145 overexpression on the expression levels of miR-145 and zeb-2 were detected with qRT-PCR and Western blotting.The changes of migration and invasion were examined using Transwell assay.Target genes of miR-145 were predicted by bioinformatics software.Dual-luciferase reporter assay were used to verify zeb-2 as a direct target of miR-145.zeb-2 siRNA was transiently transfected in SKOV3 and 3AO cells,Transwell was used to examine migration and invasion abilities.Results The migration and proliferation of SKOV3(=10.752,=0.000;=5.617,=0.005)and 3AO cells(=10.111,=0.001;=21.746,=0.000)decreased significantly after overexpression of miR-145.The results of dual-luciferase reporter assay showed that the relative luciferase activity of co-transfected miR-145 mimic and WT 3'UTR expression vectors was significantly lower than that of co-transfected mimic control and WT 3'UTR expression vectors(SKOV3:=4.572,=0.010;3AO:=3.528,=0.024).There was no significant difference in relative luciferase activity between co-transfected miR-145 mimic/MUT 3'UTR expression vector cells and co-transfected mimic control/MUT 3'UTR expression vector cells(SKOV3:=0.227,=0.831;3AO:=0.040,=0.970).Real-time quantitative PCR showed that the zeb-2 expressions in SKOV3(=1.490,=0.211)and 3AO cells(=0.114,=0.914)were not significantly different from negative control after 48 h of miR-145 overexpression.Western blot analysis showed that the expression of zeb-2 protein in SKOV3(=3.769,=0.020)and 3AO cells(=4.452,=0.011)decreased significantly compared with negative control after 72 h of miR-145 overexpression.Seventy-two hours after transfection of zeb-2 siRNA,Western blotting showed that the expression of zeb-2 protein in SKOV3(=4.660,=0.010)and 3AO cells(=4.594,=0.010)was significantly down-regulated.Transwell assay showed that the migration and invasion abilities of SKOV3(=18.655,=0.000;=18.026,=0.000)and 3AO cells(=5.500,=0.005;=8.780,=0.001)were significantly decreased.Conclusion miR-145 may inhibit the migration and invasion of ovarian cancer cells by targeting zeb-2.

Expression and clinical significance of ZEB2 and E-cadherin in nasopharyngeal carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression and clinical significance of ZEB2 and E-cadherin mRNA and protein in nasopharyngeal carcinoma (NPC) tissues.@*METHOD@#The expressions of ZEB2 and Ecadherin in 39 cases of NPC tissue and 12 cases of nasopharyngeal inflammation tissue were detected by Real-time PCR method and immunohistochemical technique. To assess their correlations with clinicopathological parameters of NPC and the interrelationship between them.@*RESULT@#Both the expression of ZEB2 mRNA and protein were higher in NPC tissues than that in inflammation tissues (P 0.05). Both the expression of Ecadherin mRNA and protein were higher in NPC tissues than that in inflammation tissues (P 0.05). In NPC tissues, the expression of ZEB2 mRNA was negative correlated with the expression of E-cadherin mRNA (r = -0.367, P < 0.05). The expression of ZEB1 protein was negative correlated with the expression of E-cadherin protein (r = -0.322, P < 0.05), the differences were both statistically significant.@*CONCLUSION@#The expression of ZEB2 was up-regulated in NPC, while the expression of E-cadherin was down-regulated, their expression was significantly negative correlated, and might be associated with metastasis of NPC, ZEB2 may promote the invasion and metastasis of NPC by inhibiting the expression of E-cadherin.

DZNep raises miR-200c expression to delay the invasion and migration of MGC-803 gastric carcinoma cells / 生理学报

The aim of the present study was to investigate the regulatory effects of histone methylation modifications on the expression of miR-200c, as well as invasion and migration of gastric carcinoma cells. Gastric carcinoma cell line, MGC-803, were treated by 2.5 μmol/L histone methyltransferase inhibitor, DZNep. The expression of miR-200c was detected by real-time quantitative PCR (qRT-PCR). The epithelial-mesenchymal transition (EMT) indicators (ZEB1/2 and E/N-cadherin), EZH2, EED, SUZ12 and H3K27me3 expressions were detected by Western blot. Cell migration and invasion abilities were detected by Transwell and scratch tests. The result showed that, compared with DMSO (control) group, DZNep significantly increased the expression of miR-200c to about 2.1 times, inhibited ZEB1, ZEB2, and N-cadherin expressions, and activated E-cadherin expression; Also, DZNep decreased the protein expressions of EZH2, EED, SUZ12 and H3K27me3; Moreover, DZNep could inhibit MGC-803 cell invasive and migrative abilities, as well as MMP9 expression. These results suggest DZNep raises miR-200c expression to delay the invasion and migration of gastric carcinoma cells, and the underlying mechanisms involve the regulations of EMT-related proteins and polycomb repressive complex 2.

Expressions of ZEB2 and C-myc in epithelial ovarian cancer and their clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression patterns of ZEB2 and C-myc in epithelial ovarian cancer (EOC) and the associations between their expressions and the pathological features of EOC.</p><p><b>METHODS</b>The expressions of ZEB2 and C-myc proteins were detected immunohistochemically in 191 cervical cancer tissues and 13 normal ovarian tissues. The relationship between ZEB2 and C-myc protein expressions and the clinicopathological features of EOC was evaluated.</p><p><b>RESULTS</b>ZEB2 positive expression ratea in EOC tissues and normal ovarian tissues were 49.2% (94/191) and 30.8% (4/13), respectively (P=0.007), and C-myc positive expression rates in the two tissues were 53.9% (103/191) and 15.4% (2/13), respectively (P=0.001). A high expression of ZEB2 was positively correlated with the pathological type of the tumor (P=0.003), FIGO stage (P=0.028), T stage (P=0.002), and N stage (P=0.04), and a high expression of C-myc was positively correlated with FIGO stage (P=0.035), histological grade (P=0.039), and T stage (P=0.002). C-myc and ZEB2 expressions were positively correlated in EOC (P<0.001), and their co-expression in EOC was significantly correlated with T stage (R=0.358, P<0.001) and FIGO stage (P=0.008).</p><p><b>CONCLUSION</b>ZEB2 and C-myc can promote the progression, invasion and metastasis of EOC, and their combined detection may assist in early diagnosis of EOC.</p>

miR-200c inhibits metastasis of breast cancer cells by targeting HMGB1 / 华中科技大学学报（医学）（英德文版）

miR-200c has been shown to regulate the epithelial-mesenchymal transition (EMT) by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software (miRanda) was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.