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Objective:To evaluate the role of ERBB2 interacting protein (Erbin)in liver tissues in blood coagulation of septic mice.Methods:Thirty SPF healthy male C57BL/6 mice and 30 Erbin knockout mice, aged 8-10 weeks, weighing 20-30 g, were divided into wild-type+ sham operation group (WT+ Sham group), wild-type+ sepsis group (WT+ SEP group), Erbin gene knockout+ sham operation group (EKO+ Sham group) and Erbin gene knockout+ sepsis group (EKO+ SEP group) by a random number table method, with 15 animals in each group. The mouse sepsis model was prepared by the cecal ligation and perforation method in anesthetized animals. Eye blood samples were collected at 24 h after surgery and liver tissues were obtained for microscopic examination of histopathological changes (by HE staining) which were scored and for determination of plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities (by colorimetry), expression of Erbin and tissue factor (TF) (by Western blot), expression of tissue plasminogen activator (t-PA) and fibrinogen (Fib)mRNA (by quantitative polymerase chain reaction), concentrations of PT, APTT, thrombin time (TT) and Fib (by automatic coagulation analyzer), and plasma TF and interleukin-6 (IL-6) concentrations (by enzyme-linked immunosorbent assay).Results:Compared with WT+ Sham group, the lung injury score was significantly increased, the expression of TF, t-PA mRNA and FGA mRNA was up-regulated, PT, APTT and TT were prolonged, the plasma Fib concentration was increased, and the activities of ALT and AST and concentrations of TF and IL-6 in plasma were increased in WT+ SEP group ( P<0.05). Compared with WT+ SEP group, the lung injury score was significantly increased, the expression of TF, t-PA mRNA and FGA mRNA was up-regulated, PT, APTT and TT were prolonged, the plasma Fib concentration was increased, and the activities of ALT and AST and concentrations of TF and IL-6 in plasma were increased in EKO+ SEP group ( P<0.05). Conclusions:Erbin in liver tissues exerts an endogenous protective effect on blood coagulation by inhibiting the up-regulation of TF expression in septic mice.
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Objective:To evaluate the relationship between Erbin and Bax/Bcl-xL-mediated cell apoptosis during sepsis-induced acute kidney injury in mice.Methods:Thirty-two SPF male wild type C57BL/6 mice, 32 SPF male Erbin (-/-) C57BL/6 mice, aged 6-8 weeks, weighing 20-30 g, were divided into 2 groups ( n=16 each) using the random number table method: wild type sham operation group (WT+ Sham group), wild type sepsis group (WT+ S group), Erbin(-/-) sham operation group (EKO+ Sham group), and Erbin(-/-) sepsis group (EKO+ S group). The sepsis model was established using the moderate cecal ligation and puncture (CLP) in anesthetized animals.The survival rates within 7 days after CLP were recorded.The serum concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10), IL-1β, creatinine (Cr), blood urea nitrogen (BUN) and lactic dehydrogenase (LDH) were determined at 24 h after CLP.Then the renal tissues were taken for assessment of renal injury which was scored and for determination of the apoptosis rate (by TUNEL) and expression of cleaved-caspase-3, Bcl-xL and Bax (by Western blot). Results:Compared with sham operation groups, the survival rates were significantly decreased, the serum concentrations of IL-1β, IL-10, TNF-α, Cr, BUN and LDH, renal injury score and apoptosis rate were increased, the expression of Bax and cleaved-caspase-3 was up-regulated, and the expression of Bcl-xL was down-regulated in sepsis groups ( P<0.05). Compared with WT+ S group, the survival rates were significantly decreased, the serum concentrations of IL-1β, LDH, TNF-α, Cr and BUN and renal injury score were increased, the serum concentration of IL-10 was decreased, the apoptosis rate of renal tissues was increased, the expression of Bax and cleaved-caspase-3 was up-regulated, and the expression of Bcl-xL was down-regulated in EKO+ S group ( P<0.05). Conclusions:Erbin can inhibit Bax/Bcl-xL-mediated cell apoptosis and is involved in endogenous protective mechanism against sepsis-induced acute kidney injury in mice.
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Objective @#To investigate the expression and clinical significance of erbb2 interacting protein (ERBIN) and PDZ domain containing 2 protein (PDZD2) in colorectal cancer tissues,as well as the effects of ERBIN and PDZD2 on the progression of colorectal cancer and the corresponding mechanisms.@*Methods @#Western blot was used to detect the expression of ERBIN and PDZD2 proteins in tumor tissues and corresponding adjacent tissues from 86 colorectal cancer patients,as well as normal human colon fibroblasts ( CCD-18Co) and human colorectal cancer cell lines ( SW480 ,HCT116 ,SW620 and HT29 ) ; SW620 and HCT116 cells were transfected with ERBIN or PDZD2 overexpression vector,and then cell proliferation,cell invasion and apoptosis were detected.The expres- sion of ERBIN or PDZD2 protein in SW620 cell was observed by immunofluorescence chemistry.Co-immunoprecip- itation assay was used to detect the interaction between ERBIN and PDZD2 proteins.@*Results @#The expression levels of ERBIN and PDZD2 proteins in colorectal cancer tissues were lower than those in adjacent tissues (P<0.01) , and the expression levels of ERBIN and PDZD2 in colorectal cancer cells were lower than those in CCD-18Co cells (P<0. 01) .The expression levels of ERBIN and PDZD2 proteins were correlated with the depth of invasion,dif- ferentiation,TNM staging and lymph node metastasis of colorectal cancer (P<0. 001) ; ERBIN or PDZD2 overex- pression reduced the proliferation and invasion of SW620 and HCT116 cells (P<0. 01) ,increased cell apoptosis (P <0. 01 ) ; ERBIN directly bound to PDZD2 ,and overexpression of ERBIN increased the expression level of PDZD2 protein.@*Conclusion @#ERBIN can inhibit the proliferation and invasion of colorectal cancer cells and pro- mote apoptosis by promoting the expression of PDZD2 protein.
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To explore the significance of Erbin expression in esophageal squamous cell carcinoma (ESCC) and its relation-ship with patient prognosis. Methods: Erbin expression in a tissue chip containing samples from 299 cases were examined using immu-nohistochemistry; in addition, the relationship between Erbin expression and clinical-pathological parameters and patient survival time were also analyzed. Cox regression was used to predict the risk of clinical-pathological parameters. The mRNA and protein expres-sion of Erbin was also determined in 25 cases with paired ESCC and normal tissues through polymerase chain reaction (PCR) and West-ern blot. Results: The expression of Erbin protein and mRNA in ESCC were significantly higher than those of normal esophageal epithe-lium adjacent to cancer (55.2% vs. 0, P<0.05). The high expression of Erbin in ESCC was closely related to TNM stage (64.9% vs. 47.3%, P=0.002) and lymph node metastasis (65.5% vs. 45.0%, P<0.001). Moreover, the high expression of Erbin in ESCC was closely related to poor prognosis (P<0.05). Conclusions: Erbin expression was increased in ESCC and was closely related to poor patient prognosis, which suggested that Erbin may be an important biomarker for the prognosis of cancer patients.
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Objective To explore the effect of Erbin deficiency in MDA-453 cells on trastuzumab(Herceptin) resist-ance .Methods The specific short hairpin RNA ( shRNA) targeting Erbin was designed and cloned into plasmid pSuppres-sor, which was subsequently transfected into MDA-453 human breast cancer cells .After being selected by G418, MDA-453 cells stably expressing Erbin shRNA were obtained and nominated as MDA-453-Erbin sh.The MDA-453 cells express-ing control shRNA ( MDA-453-NC) were used as control cells .The expression of Erbin at the protein level in MDA-453-NC and MDA-453-Erbin sh cells was analyzed by Western blotting .Cell proliferation and colony formation assays were em-ployed to investigate the effect of Erbin knockdown on the sensitivity of MDA -453 cells to trastuzumab in vitro.The levels of Erbin expression in human breast cancer tissue and normal breast tissue samples were evaluated by immunohistochemistry . Results MDA-453 cells, in which Erbin expression was stably knocked-down, were established.Deficiency of Erbin in MDA-453 cells could antagonize the anti-proliferation effects of trastuzumab in vitro.The level of Erbin expression was de-creased in some breast cancer tissue samples compared with normal breast tissue samples .Conclusion Erbin deficiency may induce the resistance of breast cancer cells to trastuzumab therapy .
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Objective To investigate the expression of Erbin in renal interstitial fibrosis (RIF) and the effect of over-expression of Erbin on transforming growth factor β1 (TGF-(β1)-induced epithelial-mesenchymal transition (EMT) in NRK52E cells. Methods In vivo, the model of renal fibrosis was induced by 5/6 subtotal nephrectomy in rat. Scr and BUN was detected and Masson staining was used to evaluate the level of renal tissue fibrosis. The location and expression of Erbin in renal tissue were detected by immunohistochemistry and Western blotting. In vitro, after NRK52E cells were treated by TGF-β1 (10 μg/L) for 72 h, immunofluorescence and Western blotting were used to obverse the expression and distribution of E-cadherin and α-SMA. The expression of Erbin mRNA and protein were detected by RT-PCR and Western blotting respectively. NRK52E cells were transiently transfected with Prk5-myc-Erbin plasmid via lipofectamine 2000, then the expressions of Erbin, E-cadherin and α-SMA were detected by Western blotting. Results (l)Compared to sham group with Scr (33.96±7.28) μmol/L and BUN (8.11±2.55) mmol/L, rats in 5/6 nephrectomy model with Scr (140.52±61.11) μmol/L and BUN (34.23±7.66) mmol/L revealed renal dysfunction. Masson staining indicated kidney interstitial fibrosis, and the expression of Erbin was significantly increased in renal tissue(2.9 folds), especially in tubular epithelia. (2)In vitro, the expressions of Erbin and α-SMA were markedly increased (2.3 folds and 2.1 folds, P<0.05, respectively) and the expression of E-cadherin was dramatically decreased in NRK52E cells stimulated by TGF-β1, which were consistent with immunofluorescence results. TGF-β1-induced E-cadherin suppression and a-SMA induction could be efficiently blocked by over-expression of Erbin (all P <0.05). Conclusions Erbin is up-regulated in renal interstitial fibrosis, and over-expression of Erbin can partly inhibit renal EMT induced by TGF-β1, which indicates Erbin playing an protective role in renal fibrosis.