circ_0023642 promotes malignant biological behaviors of gastric cancer cells via regulating miR-508-3p/ERBB4 axis / 中国肿瘤生物治疗杂志

@#Objective: To explore the influence of circ_0023642 on the proliferation and metastasis of gastric cancer GMC-803 cells by modulating miR-508-3p/ERBB4 axis. Methods: Cancer tissues and corresponding para-cancer normal tissues from 31 gastric cancer patients, who underwent surgical resection at the Second People's Hospital of Nantong City from May 2015 to March 2018, were collected for this study; meanwhile, gastric cancer cell lines (MGC-803, MKN-45 and MKN-28) were also collected. qPCR was performed to determine the expression levels of circ_0023642 and miR-508-3p in above mentioned tissues and cell lines. WB was applied to measure the expressions of ERBB4, E-cadherin, N-cadherin and Vimentin in MGC-803 cells. CCK-8 assay and Transwell assay were used to evaluate the effects of circ_0023462 and miR-508-3p expression on proliferation, migration and invasion of MGC-803 cells. Dual-luciferase reporter gene was carried out to validate whether miR-508-3p could bind to the 3' UTR of circ_0023642 and ERBB4. Results: Compared with para-cancer tissues or normal gastric mucosal cells, the expression of circ_0023642 was significantly up-regulated in gastric cancer tissues and cells lines, and the expression was highest in MGC-803 cells (P<0.05 or P<0.01). Silencing circ_0023642 dramatically decreased the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of MGC-803 cells (P<0.05 or P<0.01). Both circ_0023642 and ERBB4 could target the binding sites of miR-508-3p. Further experiments confirmed that circ_ 0023642 promoted the proliferation, migration, invasion and EMT of MGC-803 cells by sponging miR-508-3p (P<0.05 or P<0.01). Conclusion: circ_0023642, by competing ERBB4 to bind with miR-508-3p, promotes the proliferation and metastasis of gastric cancer MGC-803 cells, thus could be used as a marker for the clinical diagnosis of gastric cancer.

Protective effect of shikonin on renal fibrosis via Smad3/long-chain non-coding Erbb4-IR axis / 中国药理学通报

Aim To investigate the therapeutic effect of shikonin on unilateral ureteral obstructive nephropathy (UUO)-induced CKD mouse fibrosis model and the potential regulatory mechanism. Methods Thirty C57BL/6 mice (8 weeks old, weighing 20-22 g) were randomly divided into sham group, UUO model group, low dose shikonin group (5 mg • kg"1) , high dose shikonin group (20 mg • kg"1) and irbesartan group (20 mg • kg"1). Mice in shikonin groups were given the first drug intragastrically for one day before operation, and then were sacrificed for 10 days after continuous gavage. Creatine and urea nitrogen were detected, renal pathology was observed by HE and PAS, renal interstitial fibrosis was detected by MAS-SON, a-SMA was detected by IHC, and ot-SMA, FN, p-Smad3, Smad3 and Smad7 expression in renal tis-sues were detected by Western blot. TGF-p and long-chain non-coding Erbb4-IR were detected by RT-PCR. Results Compared with model group, serum creatinine and urea nitrogen significantly decreased after treatment with shikonin. HE showed marked improvement in renal tubular injury. PAS and MASSON staining showed decreased glycogen deposition and fibrosis. Western blot showed shikonin inhibited the activation of the key protein Smad3 in the TGF-p/Smad signaling pathway and up-regulated the expression of Smad7, while the expression of TGF-(3 and long-chain non-encoding Erbb4-IR decreased in a dose-dependent manner. Conclusions Shikonin can effectively alleviate renal fibrosis, and its mechanism may be related to the regulation of Smad3/long-chain non-coding Erbb4-IR axis.

α-humulene improves schizophrenia via NRG1/ErbB4 signaling pathway / 中草药

Objective: To study the treating mechanism of α-humulene on the schizophrenic mice. Methods: The schizophrenic models were established by dizocilpine maleate (MK801), then different concentrations of α-humulene were used to treat the mice by intragastric administration. Open-field experiment and PPI test were carried out to evaluate the spontaneous activity and sensorimotor gating function of mice. Moreover, the frontal cortex MDA, NO levels and hippocampal NRG1, ErbB4 protein expression was detected. Results: The spontaneous activity, sensorimotor gating function, MDA, NO, NRG1 and ErbB4 levels were significantly changed in model mice when compared with normal mice (P < 0.01). Compared with model group, different concentrations of α-humulene notably inhibited spontaneous activity, improved PPI value, increased NO and MDA content, down-regulated ErbB4 and NRG1 protein expression (P < 0.05, P < 0.01). Conclusion: The schizophrenia abnormal behavior of mice was improved by α-humulene via down-regulating NRG1/ErbB4 signaling pathway, so as to achieve the purpose of treating schizophrenia.

Quantification of Tyrosine Hydroxylase and ErbB4 in the Locus Coeruleus of Mood Disorder Patients Using a Multispectral Method to Prevent Interference with Immunocytochemical Signals by Neuromelanin / 神经科学通报·英文版

The locus coeruleus (LC) has been studied in major depressive disorder (MDD) and bipolar disorder (BD). A major problem of immunocytochemical studies in the human LC is interference with the staining of the immunocytochemical end-product by the omnipresent natural brown pigment neuromelanin. Here, we used a multispectral method to untangle the two colors: blue immunocytochemical staining and brown neuromelanin. We found significantly increased tyrosine hydroxylase (TH) in the LC of MDD patients-thus validating the method-but not in BD patients, and we did not find significant changes in the receptor tyrosine-protein kinase ErbB4 in the LC in MDD or BD patients. We observed clear co-localization of ErbB4, TH, and neuromelanin in the LC neurons. The different stress-related molecular changes in the LC may contribute to the different clinical symptoms in MDD and BD.

Neuregulin 1/ErbB4 signaling attenuates neuronal cell damage under oxygen-glucose deprivation in primary hippocampal neurons / 대한해부학회지

Electroacupuncture at Quchi and Zusanli points reduces cell apoptosis by activating Nrg-1 and ErbB4 signaling pathways in cerebral ischemia-reperfusion rats / 医学研究生学报

Objective The mechanism of electroacupuncture (EA) in the treatment of neurological dysfunction caused by ischemic stroke remains to be further explored. This study aimed to investigate the effects of EA at the Quchi (LI11) and Zusanli (ST36) points on the expressions of Nrg-1 and ErbB4 proteins and their correlation with the expressions of cell apoptosis-related proteins bcl-2 and Bax in cerebral ischemia-reperfusion (I-R) rats.Methods Totally 160 male adult SD rats were randomly assigned to four groups of equal number: sham operation, middle cerebral artery occlusion-reperfusion (MCAO-R) model, acupoint, and non-acupoint. After MCAO/R modeling, EA was applied at Quchi and Zusanli in the acupoint group and at ipsilateral non-acupoints below the axillary striation and apex of the coccyx in the non-acupoint group. At 3 days after treatment, the protein and mRNA expressions of Bcl-2, Bax, Nrg-1 and ErbB4 were determined by TTC, TUNEL, immunohistochemistry, Western blot, and RT-PCR.Results The neurological deficit score was significantly increased after cerebral I/R injury in the MCAO-R model, acupoint and non-acupoint groups compared with that in the sham operation group (P<0.01). EA therapy markedly reduced the neurological deficit scores (P<0.05), cerebral infarct area (P<0.05) and the number of apoptotic cells (P<0.05), up-regulated the protein expressions of Nrg-1, ErbB4 and Bcl-2 (P<0.05), and down-regulated that of Bax (P<0.05). There was a pronounced increase in the protein and mRNA expressions of Nrg-1 and ErbB4 in the acupoint group compared with the MCAO-R model and non-acupoint groups after cerebral I-R injury (P<0.01).Conclusion EA at the Quchi and Zusanli points has a neuroprotective effect in ischemic brain injury by up-regulating the expressions of endogenous Nrg-1 and its receptor ErbB4 and down-regulating those of Bcl-2 and Bax. The underlying mechanism is probably associated with the Nrg-1/ErbB4 signaling pathway.

Changes of neuregulin 1 receptor ErbB4 and their significance in FMR1 gene knockout mouse / 中华神经医学杂志

Objective To study the expression changes ofneuregulin 1 (NRGI) receptor ErbB4 in the cortex and hippocampus of fragile X mental retardation 1 (FMR1) gene knockout (KO) mice,and explore the role of ErbB4 in epilepsy susceptibility in fragile X syndrome (FXS).Methods Two weeks old male FMR1 KO mice (n=3),4 weeks old FMR1 KO mice (n=3) and 2 weeks old male wild type (WT) mice (n=3),4 weeks old WT mice (n=3) were chosen in our study.Immunohistochemical staining and Western blotting were performed to detect the ErbB4 positive neurons and protein expression in the cerebral cortex and hippocampus of each group.Double-labeling imnmunofluorescence and laser confocal microscope were used to observe the ErbB4 protein expression in the parvalbumin (PV) inter-neurons of cortex and hippocampus.Results As compared with those in the 2 and 4 weeks old WT mice,the number of ErbB4 positive neurons was significantly smaller and ErbB4 protein expressions in the cerebral cortex and hippocampus of 2 and 4 weeks old KO mice were significantly decreased (P＜0.05).The number of PV and ErbB4 co-expression neurons in cortex and CA1 and CA3 hippocampus of 2 and 4 weeks old KO mice was significantly smaller than that in the 2 and 4 weeks old WT mice (P＜0.05).Conclusion The decreased ErbB4 expression is related to the decreased number of PV positive inter-neurons in the FMR1 KO mice,which is involved in the pathogenesis of FXS epilepsy susceptibility.

Expression of ErbB4 in the neurons of Alzheimer's disease brain and APP/PS1 mice, a model of Alzheimer's disease / 대한해부학회지

Neuregulin-1 (NRG1) plays important roles in the development and plasticity of the brain, and has also been reported to exhibit potent neuroprotective properties. Although ErbB4, a key NRG1 receptor, is expressed in multiple regions in the adult animal brain, little is known about its role in Alzheimer's disease (AD). AD is characterized by progressive impairment of cognition and behavioral disturbance that strongly correlate with degeneration and death of neurons in the cerebral cortex and limbic brain areas, such as the hippocampus and the amygdala. Here, we show that the ErbB4 and phospho-ErbB4 immunoreactivities were higher intensity in the neurons of the CA1-2 transitional field of AD brains as compared to age-matched controls. Also, ErbB4 expression was increased in the neurons of the cortico medial nucleus amygdala, human basal forebrain and superior frontal gyrus of AD brains. In cerebral cortex and hippocampus of amyloid precursor protein/presenilin 1 double transgenic mice, ErbB4 immunoreactivity significantly increased in comparison to age-matched wild type control. These results suggest that up-regulating of ErbB4 immunoreactivity may involve in the progression of pathology of AD.

Expression of ErbB4 in the apoptotic neurons of Alzheimer's disease brain / 대한해부학회지

Neuregulin-1 (NRG1) signaling participates in the synaptic plasticity, maintenance or regulation of adult brain. Although ErbB4, a key NRG1 receptor, is expressed in multiple regions in the adult animal brain, little is known about its localization in Alzheimer's disease (AD) brains. We previously reported that ErbB4 immunoreactivity showed regional difference in the hippocampus of age-matched control. In the present paper, immunohistochemical characterization of the distribution of ErbB4 receptor in the hippocampus relative to pathology staging were performed in age-matched control (Braak stage 0, n=6) and AD (Braak stage I/V, n=10). Here, we found that ErbB4 immunoreactivity was significantly increased in apoptotic hippocampal pyramidal neurons in the brains of AD patients, compared to those of age-matched control subjects. In AD brains, ErbB4 immunoreactivity was demonstrated to colocalize with the apoptotic signal Bax in apoptotic hippocampal pyramidal neurons. These results suggest that up-regulation of ErbB4 immunoreactivity in apoptotic neuron may involve in the progression of pathology of AD.

An Immunohistochemical Study of ErbB4 Receptor in Alzheimer's Disease Hippocampus / 대한해부학회지

Neuregulin-1 (NRG1) signaling participates in numerous neurodevelopmental processes. Although ErbB4, a key NRG1 receptor, is expressed in multiple regions in the adult animal brain, little is known about its expression in aged human brain. We show that ErbB4 immunoreactivity was shown regional difference in the hippocampus of age-matched control and that the distribution of these molecules was altered in Alzheimer's disease (AD) brains. Immunohistochemical characterization of the distribution of ErbB4 receptor in the hippocampus relative to pathology staging were performed in age-matched control (Braak stage I/II, n=5), early AD (Braak stage III/IV, n=5) and advanced AD(Braak stage V/VI, n=10). The intensity of ErbB4 immunoreactivity was higher in neurons of the CA2 than that in CA1 or CA3 in the age-matched control. Particularly, in the early AD, ErbB4 immunoreactivity was significantly increased in the apoptotic cells of the CA2 field. In the advanced AD, ErbB4 immunostaining was more intense in the apoptotic cell of the CA2 field. In the dentate gyrus (DG), ErbB4-positive granular cell density was gradually increased in proportion to the progression of pathology of AD brains. We have also found that ErbB4 immunostaining was increased in the nucleus, suggesting that the presenilin-dependent cleavage of ErbB4 generates the soluble ErbB4 ICD (intracellular domain) that translocalized to the nucleus. Together, these results provide the immunohistochemical analysis of ErbB4 receptor in the human hippocampus staged by the progression of pathology of AD.

Expressional Change of Nitric Oxide Synthase and erbB4 in Rat Hippocampus after Seizure / 대한해부학회지

Nitric oxide has been considered to be an important modulator of the epileptic seizure response. Previous studies have mainly focused on the nitric oxide synthase (NOS) expressed in glial cells and vascular endothelial cells in the brain following seizures, while less data have been available reading the change of neuronal NOS (nNOS) produced in neurons. Polypeptide growth factors play a central role in a variety of environmentally induce structural changes in the cortex and hippocampus of adult brain. neuregulin is widely expressed in the central and peripheral nerve cells and Schwann cells, glia, oligodendrocytes and muscle cells, to control cellular proliferation, differentiation and migration. erbB family are the receptors of the neuregulin and consist of erbB2, erbB3 and erbB4. We have, therefore, investigated the change in the expression of nNOS and erbB4 in the rat hippocampus, one of the brain structures most vulnerable to seizures. Rats were injected with kainic acid (KA) and sacrificed 6 h, 1 d, 3 d and 6 d after KA administration. The expression pattern of nNOS and erbB4 was studied using reverse transcription-polymerase chain reaction analysis, NADPH-diaphorase (NADPH-d) histochemistry and immunohistochemistry. The increase in the level of nNOS reached maximal values in samples obtained 1 d after KA treatment. The optical densities of NADPH-d-positive neurons in the CA1 and dentate gyrus (DG) regions of the hippocampus were shown to have increased in samples obtained 1 d and 3 d after injection of KA. The number of NADPH-d-positive neurons in the CA1 regions of the hippocampus was shown to have decreased in samples obtained 3 d and 6 d after injection of KA. However, the number of NADPH-d-positive neurons in the DG region did not change significantly. We show that erbB4 immunoreactivity is increased in hippocampus, reaching maximal levels 3 d after KA treatment, some NOS neurons contain erbB4 protein. We propose that the survival of NOS neuron in the hippocampus after injection of KA is associated with expression of erbB4, neuregulin receptor.

Immunohistochemical Expression of c-erbB2, c-erbB3 and c-erbB4 Protein in Breast Cancer / 한국유방암학회지

Breast cancer is the third leading cause of cancer related deaths in Korean women. Members of the erbB receptor family, the EGF receptor and c-erbB2, c-erbB3 and c-erbB4, are commonly over-expressed in human breast cancer and there is a high correlation with an aggressive breast cancer phenotype and poor patient prognosis. Since the over-expression of the EGF receptor and c-erbB2 suggested that signalling of erbB receptors may contribute to the development and progression of breast cancer, we investigated the correlation of clinicopathological factors and the immunohistochemical expression of c-erbB2, c-erbB3 and c-erbB4. To determine the c-erbB immunoreactivity, we used Rabbit anti-human c-erbB2 oncoprotein (DAKO, Denmark), mouse monoclonal c-erbB3(RTJ.2 Santa Cruz) and rabbit polyclonal antibody c-erbB4(Santa Cruz) directed against each c-erbB protein by immunohistochemistry from paraffin-embedded tissue in a series of 190 women with breast cancer. About 25.8%(49 out of 190 patients) of breast cancers overxpressed c-erbB2, and 40.0%(76 out of 190 patients) and 18.9%(36 out of 190 patients) overxpressed c-erbB and c-erbB4, respectively. Poor histologic grade showed tendency of positive correlation of the positivity of c-erbB2 and 3 but without statistical significance and no correlation with c-erbB4. We observed positive correlations among c-erbB2, c-erbB3 and c-erbB4 expression.(p0.05) and lymph node status(p>0.05) were not related with c-erbB family expression. The expressions of c-erbB2, c-erbB3 and c-erbB4 showed no survival benefit or no disease free benefit compared to c-erbB family negativity in univariate analysis (Kaplan-Meier life table analysis). Our results suggest that c-erbB2, c-erbB3, and c-erbB4 may regulate the growth of breast cancer by the interactions of these family of growth factor receptor that are dependent of hormonal control.

Expression of the EGF Receptor Gene Family in Gastric Cancer

Growth factor receptors play critical roles in the regulation of normal growth and developement. Some of these molecules have been implicated in the neoplastic process as well. In this study, We examined the expression of the EGF receptor gene family in gastric carcinoma by Ribonuclease protection assay and investigated the relationship between the expression of using a mRNA and clinicopathologic parameters. Expression of EGFR mRNA was found in 34 of 59 cases (57.6%), expression of c-erbB-2 mRNA was found in 27 of 57 cases (64.9%), expression of c-erbB-3 mRNA was found in 47 of 60 cases (78.3%), and expression of c-erbB-4 mRNA was found in 39 of 59 cases (66.1%). The expression of EGFR was correlated with the size of the tumor. The expression of c-erbB-2 was correlated with the presence of endolymphatic tumor emboli, Ming's classification, and lymph node matastasis. The expression of c-erbB-3 was correlated with the macroscopic type. Coexpression of c-erbB-2 and c-erbB-3 was correlated with age. These results suggest that the production of the EGF receptor gene family by various tumor cells it may play an important role in the cellular function. Therefore, further studies are currently being carried out to clarify the role of these oncogenes in tumor behavior and gastric carcinogenesis.

Expression of EGF Receptor Family Genes and Proteins in Gastric Cacner ( EGFR , c-erbB-2 , c-erbB-3 , c-erbB-4 ) / 대한암학회지

PURPOSE: to evaluate the relationship between the expression of EGF receptor gene family and clinico-pathologic parameters. MATERIALS AND METHODS: We compared adenocarcinoma tissue with normal mucosa obtained from same patients in 60 cases. The amplifications of DNA was examined by slot blot, while the expression of mRNA by ribonuclease protection assay, and that of protein by immunohistochemistry. RESULTS: Expression of EGFR mRNA was observed in 9 of 59, that of c-erbB-2 mRNA in 8 of 57, that of c-erbB-3 mRNA in 4 of 60, and that of c-erbB-4 mRNA in 13 of 59. The expression of EGFR in expanding type showed a higher tendency than that in infiltrative type. The expression of c-erbB-2 in poorly differentiated adenocarcinoma showed a higher tendency than that in well differentiated adenocarcinoma. And expression of c-erbB-2 was correlated with presence of endolymphatic tumor emboli. No significant correlation was observed between expression of EGFR mRNA and that of its protein or amplification of its DNA. Similarly, No clear relationship between c-erbB-2 gene amplification and expression of mRNA or proteins was detected. CONCLUSION: EGFR, c-erbB-2, c-erbB-3, and c-erbB-4 were expressed in gastric adenocarcinoma in Korea. The presence of EGF receptor gene family by various tumor cells suggested that it may play an important role in adenocarcinoma. Therefore further studies are currently being carried out to clarify the role of these oncogenes in tumor behavior and gastric carcinogenesis.

Expression of ErbB4 in and apoptosis of cardiac muscle cells of rats with dilated cardiomyopathy and their intervention with neuregulin-1? / 第三军医大学学报

Objective To study the adriamycin(ADR)-induced expression of ErbB4 in and apoptosis of cardiac muscle cells of rats with dilated cardiomyopathy and their intervention with neuregulin-1?.Methods Sixty male SD rats,weighing 200-250 g,were randomly divided into ADR 1 group(n=10),ADR 2 group(n=20),ADR+neuregulin-1? group(n=20),and normal control(NC) group(n=10).Rats in ADR1 and ADR2 groups were injected with 2.0 mg/kg of ADR through the tail vein,once a week for 8 and 12 weeks,respectively.Rats in ADR+neuregulin-1? group were injected with 2.0mg/kg of ADR through the tail vein,once a week for 8 weeks,followed by neuregulin-1? [10 ?g/(kg?d)],once a day for 5 d.Rats in NC group received normal saline [2 ml/(kg?d)],once a week for 8 weeks.Expression of ErbB4 was detected by Wes-tern blotting.Lesions of cardiac muscle were scored with HE staining.Apoptosis of cardiac muscle cells was detected with the TUNEL method.Results The apoptotic index of cardiac muscle cells was significantly lower in ADR+neuregulin-1? group than in ADR 1 group(P