Cigarette smoke exposure is a major cause of chronic obstructive pulmonary disease (COPD), but the underlying molecular inflammatory mechanisms remain poorly understood. Previous studies have found that smoke disrupts cell-cell adhesion by inducing epithelial barrier damage to the adherens junction proteins, primarily E-cadherin (E-cad) and p120-catenin (p120). Recently, the anti-inflammatory role of p120 has drawn increasing attention. In this study, we demonstrate that p120 has a role in the cigarette smoke extract-induced inflammatory response, presumably by regulating NF-κB signaling activation. Mechanistically, we show that p120-mediated NF-κB signaling activation in airway epithelial inflammation is partially RhoA dependent and is independent of E-cad. These results provide novel evidence for the role of p120 in the anti-inflammatory response.

p120-catenin (p120) is known as a cadherin-associated protein that participates in tumor metastasis and invasion, as well as an anti-inflammatory mediator. Recently, its anti-inflammatory role is drawing increasing attention, but the regulatory mechanisms are still unknown. Here, we report that p120 modulated inflammatory responses partially depends on RhoA/ROCK pathway in scratch-induced injury in human bronchial epithelial cells (BECs). For the first time, we found that p120 was significantly reduced in BECs after scratching, which could induce interleukin-8 (IL-8) production through nuclear factor-κB (NF-κB) activation accompanied with IκBα phosphorylation. Over-expression of p120 3A could inhibit NF-κB activation and IL-8 mRNA expression and protein synthesis after scratching, while p120 knockdown by small interfering RNA could promote NF-κB activation and IL-8 mRNA expression and protein synthesis after scratching. Furthermore, we found that RhoA was the binding partner of p120 in BECs. Although total RhoA and p120-binded RhoA remained unchanged, the RhoA activity was increased after scratching. Chemical blockade of RhoA/ROCK signaling (Y27632) inhibited scratch-induced nuclear translocation of NF-κB p65. Over-expression of p120 3A attenuated scratch-induced RhoA activation, whereas silence of p120 significantly elevated scratch-induced RhoA activation in BCEs. Conclusively, these results indicate an anti-inflammatory effect of p120 in bronchial epithelial cells through its modulation of NF-κB signaling depending on RhoA/ROCK pathway.

The aim of the present study was to examine the effects of epithelial-mesenchymal transition (EMT) and apoptosis of renal tubular epithelial cells on the prognosis of immunoglobulin A (IgA) nephropathy. Renal biopsy tissues from 74 cases of IgA nephropathy were divided into a mild mesangial proliferation group (27 cases), a focal hyperplasia group (28 cases) and a proliferative sclerosis group (19 cases). The blood pressure, serum creatinine and 24 h urinary protein excretion of all patients were detected. To define EMT, α-smooth muscle actin (α-SMA), vimentin and collagen fibers were assessed. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). The blood pressure, serum creatinine and 24 h urinary protein excretion of patients with IgA nephropathy altered with increasing pathological grade. All clinical indices of patients in the proliferative sclerosis group were higher than those of the other two groups, and the 24 h urinary protein excretion of the focal hyperplasia group was statistically higher than that of the mild mesangial proliferation group. The expression of tubular interstitial α-SMA, vimentin and collagen fibers increased with the pathological grade and was closely correlated with clinical indices, including collagen fibers and 24 h urinary protein excretion. TUNEL-positive cells increased with the exacerbation of pathological changes. The EMT and apoptosis of renal tubular epithelial cells reflected the clinical severity of IgA nephropathy. α-SMA, vimentin and the apoptotic index may be used as important markers for evaluating the prognosis of IgA nephropathy.

p120-Catenin (p120) is an adherens junction protein recognized to regulate cell-cell adhesion. Emerging evidence indicates that p120 may also play an important role in inflammatory responses, and the regulatory mechanisms are still unknown. In the present study, we showed that p120 was associated with airway inflammation. p120 downregulation induced nuclear factor- κ B (NF- κ B) activation, accompanied with I κ B α degradation, p65 nuclear translocation, and increased expression of interleukin-8 (IL-8) in lipopolysaccharide (LPS)- treated C57BL mice and human bronchial epithelial cells (BECs). Moreover, we first found that p120 directly coprecipitated with RhoA in BECs. After LPS stimulation, although total RhoA and p120-bound RhoA were unchanged, RhoA activity was increased. Y27632, a ROCK inhibitor, could partially inhibit nuclear translocation of p65. Overexpression of p120 inactivated RhoA and NF- κ B in BECs, whereas p120 loss significantly increased RhoA activity, p65 nuclear translocation, and IL-8 expression. Taken together, our study supports the regulatory role of p120 in airway inflammation and reveals that p120 may modulate NF- κ B signaling partially through RhoA.

A case with an Alpha-fetoprotein-producing（AFP-producing） carcinoma originating from the rectum was described.A 41-year-old man,who underwent a palliative surgery for rectal carcinoma,was diagnosed with occupying liver lesions and a remarkable AFP elevation（3484.61 ng/mL）,and the AFP declined obviously 10 days after the palliative surgery.So we carried out a biopsy of the liver lesions.The histopathology was reported as low differentiation adenocarcinoma.The immunohistochemistry of the tumor cells via liver biopsy showed:Villin,CDX-2 was positive,Glypican-3 was partial positive,CK7,CK20,AFP,Hepatocyte were all negative.The initial histopathology was reported as an AFP-producing rectum adenocarcinoma with liver metastasis,which was a rare disease.So far,only 17 reports,none has been reported in China.Then,we summarize the characteristic of the disease:diagnosed with hepatic metastasis,raised serum AFP and a poor outcome,in addition to primary symptoms.This kind of disease is highly malignant.

Objective To investigate the expression of p120ctn,Kaiso and matrilysin proteins in non-small cell lung cancer(NSCLC) tissues,and to analyze their effect on tumorigenesis and development.Methods The expression of p120ctn,Kaiso and matrilysin proteins were detected using immunohistochemistry in 16 cases of bronchiectasis and 98 cases of NSCLC,including 52 cases of lung squamous cell carcinoma and 46 cases of lung adenocarcinoma.The relationship between p120ctn,Kaiso,matrilysin and clinicopathologic characteristics was analyzed.Results The abnormal expression rates of p120ctn were 74.5%,6.3% in NSCLC and bronchiectasis specimens respectively,and the difference showed statistical significance(P<0.001).The ectopic expression rates of Kaiso in lung cancer cells,goblet cells,ciliated cells and basal cells were 63.3%,6.3%,6.3%,37.5%,respectively,and the difference of ectopic expression rates between lung cancer cells and goblet cells or ciliated cells was statistically significant(P<0.001).The abnormal expression of p120ctn and cytoplasmic matrilysin expression in patients with NSCLC were both related to histological type(P<0.05).The ectopic expression of Kaiso in patients with NSCLC was related to the grade of differentiation(P<0.05).Nuclear Kaiso expression was correlated with cytoplasmic matrilysin expression(r=-0.23,P=0.02).Conclusion P120 may play a role in the NSCLC development through co-localization with Kaiso in the cytoplasm,relieving the repression of Kaiso to its target gene,and promoting matrilysin expression.

Objective To explore the role and expression of beta-catenin(β-cat) and retinoic acid receptor beta(RARβ) in the airway epithelial squamous metaplasia of vitamin A deficiency(VAD) rats.Methods SD ablactated male rats were randomized into normal group,VAD for 9 weeks group and VAS for 13 weeks group.The rat model of squamous metaplasia was established by feeding with vitamin A deficiency diet.Morphological changes of airway epithelium were observed under microscope.Immunohistochemical assay was used for the examination of β-cat and RARβ expression in the rat airway epithelium,and in situ hybridization method was performed to investigate the expression of β-cat mRNA.Results Hyperplasia and squamous metaplasia of the airway epithelium occurred in VAD groups.Compared with the normal control group,β-cat protein expression in cytoplasm was increased(P0.001),the expression of β-cat mRNA remained unchanged,and the expression of RARβ was statistically decreased(P0.01) in VAD groups.Conclusion In the process of airway epithelium squamous metaplasia caused by VAD,the expression of β-cat and RARβ increases,but the increase of β-cat expression has no correlation with β-cat mRNA.The mechanism is probably related with the decrease of RARβ,which amplifies β-cat/TCF signal and induces squamous metaplasia.

P120-catenin (p120), a prototypic member of a subfamily of Armadillo repeat domain (Arm domain) proteins, not only participates in cell–cell adhesion, but also mediates inflammatory responses in the skin. In the present study, we demonstrated the effect of p120 on lipopolysaccharide (LPS)-induced inflammatory responses in human bronchial epithelial cells (BECs). We first confirmed that p120 expression was significantly reduced after LPS stimulation in BECs, the p65 subunit of nuclear factor-κB (NF-κB) nuclear translocation was promoted and NF-κB activity was rapidly induced. Moreover, the expression level of interleukin-8 (IL-8) increased after LPS treatment. Over-expression of p120 attenuated LPS-stimulated NF-κB reporter gene expression and IL-8 mRNA expression and protein synthesis. On the contrary, transfection with p120 small interfering RNA (siRNA) significantly elevated LPS-stimulated NF-κB transcriptional activity, p65 nuclear translocation and IL-8 expression. Collectively, these results indicate an anti-inflammatory effect of p120 in BECs, through its modulation of NF-κB signaling.

Nuclear factor-κB (NF-κB) plays a central role in the development of bleomycin (BLM) lung toxicity, but the regulatory mechanisms are still unknown. In the present study, we investigated the cytotoxic effect of BLM on cultured human bronchial epithelial cells (BECs) and first confirmed that BLM induced the transcriptional activation of NF-κB signaling in BECs. We also found that BLM activated Akt (protein kinase B, PKB) and increased the phosphorylation level of glycogen synthase kinase 3β (GSK3β). GSK3β is known to be a key downstream target of Akt, and LY294002, the PI3K (phosphatidylinositol 3-kinase)/Akt inhibitor, which promoted the dephosphorylation of GSK3β, significantly attenuated BLM-induced NF-κB activation. Next, we further observed that constitutively active GSK3β stabilized the inhibitor of NF-κB (IκBα), inhibited p65 nuclear translocation and partially blocked BLM-induced NF-κB activation. Importantly, a co-immunoprecipitation assay revealed that GSK3β formed a complex with IκBα, while GSK3β phosphorylation caused by BLM led to their dissociation. These results suggest that BLM can induce the activation of NF-κB signaling in BECs and this process is tightly associated with the phosphorylation status of GSK3β, implying a possible regulatory mechanism of NF-κB signaling in BECs during the toxic lung injury induced by BLM.

Cigarette smoke is known to have various injurious and cytotoxic effects on alveolar epithelial cells. However, the mechanism about the effects caused by cigarette smoke on alveolar epithelial cells remains unclear.In the present study, we first validated that cigarette smoke extract (CSE) impaired the viability of alveolar epithelial cells (A549 cells) and resulted in some morphological changes. Next, we found that glycogen synthase kinase 3 beta (GSK3 beta) was highly expressed in A549 cells, and CSE significantly inhibited GSK3 beta by reducing GSK3 beta expression and increasing inactive phosphorylated GSK3 beta. It was also observed that CSE promoted beta-catenin accumulation and nuclear translocation, and further activated beta-catenin/TCF signaling. Finally, we demonstrated that GSK3 beta over-expression promoted the degradation of P-catenin and abolished beta-catenin/TCF transcriptional activity that was induced by CSE in alveolar epithelial cells. These results suggest that CSE induces the activation of beta-catenin/TCF signaling through inhibiting GSK3 beta, implying a possible mechanism responsible for the injurious and cytotoxic effects on alveolar epithelial cell caused by cigarette smoke. (c) 2009 Elsevier Ireland Ltd. All rights reserved.

蓝儒竹医师:首先简要介绍一下患者情况.男性患者,55岁,因"左侧腰部胀痛2个月"于2007年4月入院.不伴有尿频、尿急、尿痛及肉眼血尿等,亦无畏寒发热.既往无特殊疾病.体检:腹部平软,左肾区轻度叩击痛.

Gleason分级系统是由美国病理学家Donald F.Gleason在总结了4000多例前列腺癌标本的组织学特点和临床特征的基础上,于1966年首先提出的.经过数十年的临床应用,表明该系统能较好地反映肿瘤的生物学特征和预后.在1993年由WHO推荐为前列腺癌的标准病理分级系统[1].Gleason分级系统目前已成为前列腺癌最常用的组织病理学分级系统.本文就该分级系统的基本观念、临床价值以及新近的相关争论进行概述.

Objective To investigate the roles of IQ motif containing GTPase activating protein 1(IQGAP1) and cell division cycle 42 protein(Cdc42) in the repair of the injured airway epithelial cells(AECs) at different stages.Methods Twenty-four Kunming mice were randomly divided into four groups as following:3,7,60-day formaldehyde(FA) inhalation group and the control group.The mice were exposed to gaseous FA of 5 mg/m3 for 4h/day for 3,7,and 60 days.The expressions and locaiizations of IQGAP1 and Cdc42 protein in the AECs were observed during the repair.Results IQGAP1 of the 3-and 7-day groups significantly increased(P0.01),but decreased at the 60th day.Cde42 protein obviously decreased after 3-day inhalation of FA(P0.01),but enhanced again at the end of 7th day.IQGAP1 localized around the plasma membrane and Cdc42 protein mainly distributed uniformly in the cytoplasm of AECs.In 3-and 7-day treated groups they were co-localized at the cell leading edge,and in 60-day treated group their localizations were similar to the control.Conclusion During the injury and the repair after the injury,the expression and the localization of IQGAP1 and Cdc42 protein are dynamically changed in the AECs.They may play a key role in the repair process of airway epithelium injury.

For a preliminary study of the role of β-catenin/Tcf signaling in squamous differentiation of airway（tracheobronchial） epithelial cells,a stable mutant of β-catenin was transfected into primar-ily cultured porcine airway epithelial cells.Western blotting revealed that exogenous protein was ob-served in large quantity in cytoplasm and nucleus.When co-transfected with Tcf luciferase reporter plasmids,β-catenin mutant increased the reporter’s transcriptional activities.However,mRNA ex-pression of a squamous differentiation marker,small proline-rich protein（SPRP）,was not elevated,as shown by reverse transcription-polymerase chain reaction.These findings suggest that β-catenin/Tcf signaling may not be directly involved in the squamous differentiation of porcine airway epithelial cells.

The process of injury and repair involves spreading, migration and cell proliferation. The functions of Rho GTPases and their effector IQGAP1 are poor known in this process of airway epithelium. In the present study, we employed a widely used in vitro model by scratching a monolayer of BECs. We found that scratching induced decreasing of the GTP-bound Rac1 and Cdc42, but increasing the amounts of IQGAP1 at different time points. Next, we confirmed that IQGAP1 interacted with the constitutively active Rac1 (Rac1V12) and Cdc42 (Cdc42V12) rather than the dominant negative Rac1 (Rac1N17) and Cdc42 (Cdc42N17). Over-expressions of wild type (WT) IQGAP1 and its mutant (T1050AX2), which was defective to interact with Rho GTPases, induced translocation of β-catenin from the cytoplasm into the nucleus. These results activated Tcf/Lef and increased the expression levels of its target genes of c-myc and cyclin D1. Likewise, the amounts of c-myc and cyclin D1 increased after scratching. Our results suggested that IQGAP1 mediated cell proliferation through activating Tcf in a manner independent of Rac1 and Cdc42 in wound repair of BECs.

Glycogen synthase kinase 3beta (GSK3beta) regulates numerous signaling pathways that control a wide range of cellular processes, including cell proliferation, differentiation, apoptosis and metabolism. We report a novel function of GSK3beta: It interacts with the inhibitor-of-apoptosis protein (IAP) survivin to modulate its expression, thus regulating apoptosis in human lung cancer cells. A co-immunoprecipitation assay revealed that GSK3beta can bind survivin. Activation of GSK3beta induced translocation of survivin from the cytoplasm to the nucleus, resulting in G1 cell-cycle arrest and apoptosis, as well as sensitization to the chemotherapeutic drug doxorubicin. In contrast, inactivation of GSK3beta, either by transfection of a dominant-negative mutant inhibitor DN-GSK3beta or with selective inhibitor LiCl, increased cytoplasmic survivin expression, leading to cell-cycle progression and resistance to apoptosis. These results identify a pro-apoptotic role for GSK3beta in cancer cells, through its modulation of survivin in subcellular redistribution. This new role suggests that there is a potential for pharmacologic activation of GSK3beta to enhance treatment of cancer patients, including those with resistance.

3375 GSK3β, a regulator of numerous signaling pathways, is involved in a wide range of cellular processes, including cell proliferation, differentiation, apoptosis and metabolism. Here, we report a novel function of GSK3β to interact with survivin, a member of inhibitor-of-apoptosis protein (IAP) family, and to modulate its expression in regulating apoptosis in human lung cancer cells. Co-immunoprecipitation assay revealed that GSK3β was able to bind survivin, and activation of GSK3β induced translocation of survivin from the cytoplasm to the nucleus. The GSK3β-mediated nuclear localization of survivin resulted in G1 cell-cycle arrest and apoptosis in a human lung adenocarcinoma cell line A549. In contrast, inactivation of GSK3β, either by transfection of a dominant-negative mutant inhibitor DN-GSK3β or by selective inhibitor LiCl, increased cytoplasmic survivin expression leading to cell cycle progression and proliferation. These results identify a pro-apoptotic role of GSK3β through modulating survivin expression in lung cancer cells and suggest a potential of pharmacologic activation of GSK3β in therapeutics of lung cancer patients.

BACKGROUND & OBJECTIVE p53 gene plays an important role in regulating cell cycle, maintaining completeness of cellular genomes, inducing cell differentiation and apoptosis. p53 gene mutation occurs usually in gliomas, especially astrocytomas. This study was to investigate p53 gene mutation and its correlation to the development of glioma. METHODS p53 gene mutation in 41 specimens of human gliomas was analyzed by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing. RESULTS p53 gene mutations were found in 17 (41.5%) of the 41 specimens of gliomas. The p53 mutations located on exons 5-8: 7 cases (41.2%) on exon 5; 1 (5.9%) on exon 6; 4 (23.5%) on exon 7; 5 (29.4%) on exon 8. The mutation rate of p53 gene was significantly higher in grade III-IV gliomas than in grade I-II gliomas (P<0.01). DNA sequencing indicated that the p53 gene mutations were point mutations and deletions of exons in the 17 positive cases, including 13 cases (76.5%) of missense mutation, 2 cases (11.8%) of samesense mutation, and 2 cases (11.8%) of frame shift mutation. G_A and A-->G were major base mutations (55.6%). CONCLUSIONS p53 mutations always locate on exons 5 and 8. Missense mutation is major mutant type of p53 gene. p53 mutation plays an important role in the development and malignant transformation of human gliomas.

The re-epithelialization process of the airway involves spreading and migration followed by cell proliferation. Scaffold IQ domain GTPase-activating protein (IQGAP1), an effector of Rho GTPases, is a key component in a series of cell processes, although its exact mechanism in injury and repair of the airway is still unclear. In this study, we utilized a widely used model in vitro by scratching bronchial epithelial cells (BECs). At different time points after scratching, the amounts of IQGAP1 in mRNA and protein were greater than that in the control. PKCepsilon-mediated phosphorylation of IQGAP1 was involved in the process of injury and repair. The overexpression of PKCepsilon or treatment with phorbol-12-myristate-13-acetate (the PKC activator) promoted wound closure. On the contrary, the group treated with GF109203X (the PKC inhibitor) had the opposite effect. Scratching or overexpression of IQGAP1 induced increasing amounts of total beta-catenin and the transposition of beta-catenin from the cytoplasm into the nucleus. These results activated the T cell factor/lymphoid enhanced factor and induced expression levels of its target genes of c-myc and cyclin D1. The reduction of IQGAP1 by the transfection of small interference RNA of IQGAP1 attenuated these effects and directly impaired the scratching-induced wound closure. Taken together, our results suggest that IQGAP1 promotes cell proliferation and phosphorylation of IQGAP1 is involved in the process of wound closure in BECs.

Emerging evidence suggests that tubular epithelial-mesenchymal transition (EMT) is an important factor in renal tubulointerstitial fibrosis. This review describes the recent findings in the context of the tubular epithelial-mesenchymal transition process and discusses the possible mechanisms involved.