OBJECTIVE: To explore the genetic basis for a girl with distinctive facial features, epilepsy, intellectual disability, chronic constipation and hypopigmentation of neck and upper extremities. METHODS: Whole exome sequencing was carried out for the proband. Candidate variant was verified by Sanger sequencing. RESULTS: The proband was found to harbor a heterozygous nonsense c.586G>T (p.Glu196*) variant of the ZEB2 gene, which was unreported previously. The variant was not detected in either parent. CONCLUSION The ZEB2 gene c.586G>T (p.Glu196*) variant probably underlay the Mowat-Wilson syndrome in this patient. Hypopigmentation in the neck and upper extremities may be related to Mowat-Wilson syndrome. Prenatal diagnosis was recommended for subsequent pregnancies.
Facies ; Female ; Hirschsprung Disease ; Humans ; Hypopigmentation ; Intellectual Disability/genetics* ; Microcephaly ; Pregnancy ; Zinc Finger E-box Binding Homeobox 2/genetics*

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.
Facies ; Genetic Variation ; Heterozygote ; Hirschsprung Disease ; genetics ; Humans ; Intellectual Disability ; genetics ; Microcephaly ; genetics ; Whole Exome Sequencing ; Zinc Finger E-box Binding Homeobox 2 ; genetics

BACKGROUND: microRNAs play an important role in the development and biological phenotype of lung cancer. The present study was to investigate miR-367-3p level in non-small cell lung cancer (NSCLC) patients and its biological function of NSCLC cells. METHODS: Twenty-two patients with NSCLC (13 cases of adenocarcinoma and 9 cases of squamous carcinoma) admitted to our hospital and treated by surgery were included. During the operation, cancer tissue, paracancerous tissue and 5 mL peripheral blood were collected. Meanwhile, 22 healthy controls were selected and 5 mL peripheral blood was taken. Real-time PCR was applied to detected the expression of miR-367-3p in cancer tissues, peripheral blood of patients with NSCLC and healthy controls. miR-367-3p was detected in lung cancer cell lines (A549) and normal bronchial epithelial cells (BEAS-2B). The proliferation and invasion ability of A549 cells before and after infection were detected by MTT and Transwell assay after transfection with exogenous miR-367-3p. The downstream target gene of miR-367-3p was analyzed by bioinformatics. Zinc finger E-box binding homeobox 2 (ZEB2) was detected by Real-time PCR and Western blot. RESULTS: miR-367-3p in cancer tissues of 22 NSCLC patients was lower than corresponding normal tissues (P<0.05), and the serum miR-367-3p level in healthy subjects was higher than NSCLC subjects (P<0.05). The area under the receiver operating characteristic (ROC) curve of NSCLC was 0.95 (95%CI: 0.89-1.00) and 0.85 (95%CI: 0.74-0.97) respectively; The proliferation and migration ability of lung cancer cell line A549 transfected with exogenous miR-367-3p decreased significantly (P<0.05); Bioinformatics predicted that the downstream target of miR-367-3p was ZEB2 and up-regulating miR-367-3p expression, ZEB2 gene was decreased (P<0.05). The Cancer Genome Atlas (TCGA) data analysis showed that there was no significant difference in overall survival (OS) and disease free survival (DFS) between ZEB2 high expression group and low expression group (P>0.05). ZEB2 expression was positively correlated with infiltration of B lymphocytes (r=0.32, P<005), CD8⁺ T cells (r=0.44, P<005), CD4⁺ T cells (r=0.46, P<005), macrophages (r=0.65, P<005), neutrophils (r=0.73, P<005) and dendritic cells (r=0.71, P<005) in NSCLC. CONCLUSIONS The expression of miR-367-3p is down regulated in NSCLC patients and participates in the biological process of proliferation and invasion of NSCLC by targeting ZEB2 gene.
Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Cell Proliferation ; MicroRNAs/genetics* ; A549 Cells ; Zinc Finger E-box Binding Homeobox 2/genetics*

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.
DNA Copy Number Variations ; Facies ; Hirschsprung Disease ; Humans ; Intellectual Disability/genetics* ; Microcephaly/genetics* ; Zinc Finger E-box Binding Homeobox 2/genetics*

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.
Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Female ; Humans ; MicroRNAs ; genetics ; Neoplasm Invasiveness ; Ovarian Neoplasms ; pathology ; Zinc Finger E-box Binding Homeobox 2 ; genetics

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.
Biomarkers, Tumor ; Breast Neoplasms ; genetics ; metabolism ; Cell Movement ; genetics ; Epithelial-Mesenchymal Transition ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; HEK293 Cells ; HMGB1 Protein ; genetics ; Homeodomain Proteins ; biosynthesis ; Humans ; MicroRNAs ; genetics ; Neoplasm Invasiveness ; genetics ; Neoplasm Metastasis ; genetics ; pathology ; Repressor Proteins ; biosynthesis ; Transcription Factors ; biosynthesis ; Zinc Finger E-box Binding Homeobox 2 ; Zinc Finger E-box-Binding Homeobox 1

OBJECTIVETo 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.

METHODSThe 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.

RESULTSZEB2 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).

CONCLUSIONZEB2 and C-myc can promote the progression, invasion and metastasis of EOC, and their combined detection may assist in early diagnosis of EOC.

Disease Progression ; Female ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Neoplasms, Glandular and Epithelial ; genetics ; metabolism ; Ovarian Neoplasms ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Zinc Finger E-box Binding Homeobox 2