miR-124 regulates proliferation, apoptosis, migration and invasion of HCC cells via targeting ZEB2 / 中国药理学通报

Aim To observe the effeet of changes in miR-124 expression on the proliferation, apoptosis, migration and invasion of HCC eells and its mecha¬nism.Methods The expression levels of miR-124 and ZEB2 were deteeted in HepG2 eells.CCK8, flow cytometry, Edu and Fran swell were used to deteet the effeets of miR-124 and ZEB2 on eell proliferation, ap¬optosis, migration and invasion.Dual lueiferase and target genes were used to prediet the targeting relation¬ship between miR-124 and ZEB2.The effeet of miR- 124 and ZEB2 on proliferation, apoptosis, migration and invasion-related protein expression was deteeted by Western blot.Results The expression of miR-124 in HepG2 eells was lower than that in normal liver eells L-02, while ZEB2 and miR-124 showed the opposite trend.The results of bioinformaties prediction and dual lueiferase showed that the expression of ZEB2 was neg¬ atively correlated with the expression of miR-124.Overexpression of miR-124 and silencing ZEB2 signifi¬cantly inhibited cell proliferation activity, migration and invasion ability compared with the control group; silencing miR-124 and overexpression of ZEB2 signifi¬cantly promoted cell proliferation activity, migration and invasion ability.Western blot results showed that overexpression of miR-124 and silencing ZEB2 signifi¬cantly promoted Bax expression and inhibited Bcl-2, PCNA, MMP2 and MMP9 expression levels.Silencing miR-124 and overexpression ZEB2 were the opposite.Conclusion miR-124 could negatively regulate the effects of ZEB2 on the proliferation, migration and in¬vasion of HCC cells.

miR-9 regulates the malignant biological behaviors of small cell lung cancer by targeting zinc finger E-box binding homeobox 2 and its possible mechanism / 中国肿瘤生物治疗杂志

@#[Abstract] Objective：To explore the regulatory effect of miR-9 on biological behaviors of small cell lung cancer (SCLC) cells by targeting zinc finger E-box binding homeobox 2 (ZEB2), and to analyze the role of miR-9 in SCLC and its possible mechanism. Methods: qPCR, WB and immunohistochemistry methods were used to detect the mRNA and protein expressions of ZEB2 in cancer tissues and corresponding adjacent tissues of 67 SCLC patients who received surgical treatment at the Department of Oncology, Fourth Hospital of Hebei Medical University from February 2018 to November 2019. TargetScan was used to predict the potential target gene of miR-9, which was later verified by Dual luciferase reporter gene assay, qPCR and WB methods. CCK-8 method, Flow cytometry and Transwell experiment were used to detect the effect of miR-9 and ZEB2 over-expression on the biological behaviors of NCI-H446 cells, and WB was used to detect the protein expressions of E-cadherin, N-cadherin and Vimentin in cells. NCI-H446 cells overexpressing miR-9 were used to construct SCLC nude mouse xenograft model, and the effect of miR-9 on the growth of xenografts was observed. Results: The mRNA and protein expression levels of ZEB2 in SCLC tissues were significantly higher than those in adjacent tissues (P<0.01). There is a potential binding site on the 3' UTR of ZEB2 to bind with miR-9. Compared with the control group, the mRNA and protein expression levels of ZEB2 in NCI-H446 cells of the miR-9 over-expression group were significantly reduced (P<0.01); the proliferation, migration and invasion abilities of NCI-H446 cells were significantly suppressed (P<0.05 or P<0.01), and the expression of EMT protein was reduced; However, simultaneous over-expression of ZEB2 could reverse above effects. In in vivo experiments, the size and weight of transplanted tumors in the miR-9 over-expression group were significantly lower than those in the control group (P<0.05 or P<0.01). The expression of ZEB2 protein in the tumor tissues of nude mice in the miR-9 overexpression group was significantly lower than that in the control group (P<0.01). Conclusion: miR-9 can inhibit the biological behaviors of SCLC cells and the growth of NCI-H446 transplanted tumors in nude mice by targeting and regulating ZEB2.

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 of CtBP1, Zeb1, Zeb2 and E-cadherin in cholangiocarcinoma and their clinical significance / 临床与实验病理学杂志

Purpose To investigate the expression of transcriptional suppressor CtBP1,Zeb1,Zeb2 and their target gene E-cadherin,and their significance in cholangiocarcinoma.Methods The expression of CtBP1,Zeb1,Zeb2 and E-cadherin proteins in cholangiocarcinoma and the paired non-neoplastic tissue array were detected by the immunohistohemical staining.Results The positive rates of CtBP1 expression in cholangiocarcinoma and the paired non-neoplastic tissue were 44.44％ and 17.86％,these of Zeb2 were 34.92％ and 10.71％,and these of E-cadherin were 50.79％ and 100％,respectively.The differences between the groups were statistically significant (all P ＜ 0.05).There was only one case with expression of Zeb1 in cholangiocarcinoma,but no expression in the paired non-neoplastic tissue.CtBP1 was correlated with the degree of differentiation of cholangiocarcinoma (P ＜ 0.05).Ecadherin was related to the differentiation degree,and distant metastasis of cholangiocarcinoma (all P ＜ 0.05).The E-cadherin expression was negatively correlated with CtBP1 and Zeb2 (r =-0.034,-0.029,all P ＜ 0.05).The Zeb2 expression was positively correlated with CtBP1 (r =0.228,P =0.005).Conclusion CtBP1,Zeb2 and E-cadherin express abnormally in cholangiocarcinoma.CtBP1,Zeb2 may be involved in the regulation of E-cadherin expression.Joint detection of CtBP1 and Ecadherin is expected to be a reference index to evaluate the malignant biological behavior of cholangiocarcinoma.

Expression and significance of BCL-6 and ZEB2 in breast cancer / 临床与实验病理学杂志

Purpose To explore the biological significance of BCL-6 and ZEB2 in invasion,metastasis and prognosis of breast cancer.Methods The expressions of BCL-6,ZEB2 protein and mRNA were detected respectively in 228 cases of breast cancer and 80 cases of breast benign lesions by immunohistochemical SP two-step staining and situ hybridization.Result The expression levels of BCL-6,ZEB2 protein and mRNA in breast cancer tissues were significantly higher than in breast benign lesions (P ＜ 0.05).The expressions of BCL-6 was positively correlated with tumor size,lymphatic metastasis,histological grade,TNM staging and HER-2 expression (P ＜ 0.05).The expressions of ZEB2 was positively correlated with tumor size,lymphatic metastasis,TNM staging and HER-2 expression (P ＜ 0.05).The overall survival and relapse-free survival of BCL-6 and ZEB2 positive expression were significantly less than the negative expression (P ＜ 0.01).Conclusion The BCL-6 and ZEB2 are closely correlated with the evolution process of breast cancer,which may become important means for monitoring and warning the metastasis,invasion,and prognosis of breast cancer.

Expression of miR-200c in endometrial carcinoma and its correlation with ZEB2 protein / 实用医学杂志

Objective To investigate the expression of miR-200c in endometrial carcinoma and its correlation with ZEB2 protein. Methods The expression levels of miR-200c and ZEB2 gene and protein in tissues were detected. The expression of miR-200c in endometrial carcinoma RL95-2 cell line was inhibited by using antisense oligonucleotides and its effect on cell invasiveness was tested. The expressions of ZEB2 gene and protein in cells were detected. Results The expression levels of miR-200c and ZEB2 mRNA and protein in endometrial tissues were significantly higher than those in adjacent tissues and control group (P < 0.05). The average penetrating number in antisense miR-200c transfected group was significantly less than negative control group and liposome group (P < 0.05). The expression levels of ZEB2 gene and protein in antisense miR-200c transfected group were lower than the negative control group and liposome group (P < 0.05). Conclusion The expression of mRNA-200c in endometrial carcinoma was high and it might be promoting tumor cell invasion and metastasis by regulating ZEB2.

A case of Mowat-Wilson syndrome with developmental delays and Hirschsprung's disease

Mowat-Wilson syndrome is an extremely rare genetic disease that is characterized by intellectual disability, facial dysmorphism, Hirschsprung's disease, and other congenital anomalies. This disorder is caused by heterozygous mutations or deletions in the zinc finger E-box-binding homeobox-2 gene (ZEB2). Thus far, approximately 200 cases of Mowat-Wilson syndrome have been reported worldwide. In Korea, only one case with a 2q22 deletion, which also affects ZEB2, has been previously reported. Here, we describe a patient with Mowat-Wilson syndrome who presented with developmental delays, typical facial dysmorphism, and Hirschsprung's disease. Molecular analysis of ZEB2 identified a novel heterozygous mutation at c.190dup (p.S64Kfs*6). To our knowledge, this is the second report of a Korean patient with Mowat-Wilson syndrome that has been confirmed genetically.

Síndrome de Mowat – Wilson, primer caso reportado en Colombia: Importancia de un estudio ampliado por genética / Mowat - Wilson syndrome, first case reported in Colombia: Importance of expanded by genetic study

El Síndrome de Mowat - Wilson (SMW), es una rara enfermedad genética con prevalencia desconocida, hasta el año 2010 habían sido descritos 180 casos en la literatura mundial indexada. Los pacientes con SMW presentan un fenotipo característico dado por: frente amplia y abombada, hipoplasia mediofacial, hipertelorismo ocular, nariz y columnela prominentes, asociado a compromiso neurológico (epilepsia, retardo en neurodesarrollo y mental) y enfermedad de Hirschsprung, que ha sido descrita solo en algunos casos. (1,2) El SMW se origina por mutaciones puntuales en el gen ZEB2 y hasta en el 17% de los casos se presenta por deleciones submicroscópicas que comprometen la región cromosómica 2q22.3 donde se localiza este gen (3). El gen ZEB2 es determinante en la diferenciación de las células derivadas de la cresta neural y sistema nervioso central lo cual explicaría el fenotipo neurológico. Los autores describimos el primer caso de SMW en población Colombiana, causado por microdeleción de novo de la región cromosómica 2q22.2, resaltando así la importancia de un completo estudio citogenético – molecular para pacientes con alta sospecha de síndromes de microdeleción (1,3).

Mowat – Wilson Syndrome (MWS), is a rare disease the prevalence is currently unknown approximately 180 cases had been reported until 2010. MWS patients exhibit a characteristic phenotype given by, high forehead, frontal bossing, midface hypoplasia, ocular hypertelorism, prominent nose and columella associated with neurological involvement (epilepsy and moderate to severe intellectual deficiency) and Hirschsprung disease, which has been described only in some cases (1,2). MWS is caused by mutations in the gene ZEB2 and up to 17% of the cases presented by submicroscopic deletions that compromise 2q22.3 chromosomal region where this gene is located (3). ZEB2 is critical for the differentiation of cells derived from the neural crest and central nervous system which would explain the neurological phenotype. We describe the first case of MWS in Colombian caused by de novo microdeletion of chromosome 2q22.2 region, We highlighted the importance of a complete cytogenetic and molecular study in patients with high suspected of microdeletion syndromes (1,3).