CARMA3 is overexpressed in colon cancer and regulates NF-κB activity and cyclin D1 expression

CARMA3 was recently reported to be overexpressed in cancers and associated with the malignant behavior of cancer cells. However, the expression of CARMA3 and its biological roles in colon cancer have not been reported. In the present study, we analyzed the expression pattern of CARMA3 in colon cancer tissues and found that CARMA3 was overexpressed in 30.8% of colon cancer specimens. There was a significant association between CARMA3 overexpression and TNM stage (p=0.0383), lymph node metastasis (p=0.0091) and Ki67 proliferation index (p=0.0035). Furthermore, knockdown of CARMA3 expression in HT29 and HCT116 cells with high endogenous expression decreased cell proliferation and cell cycle progression while overexpression of CARMA3 in LoVo cell line promoted cell proliferation and facilitated cell cycle transition. Further analysis showed that CARMA3 knockdown downregulated and its overexpression upregulated cyclin D1 expression and phospho-Rb levels. In addition, we found that CARMA3 depletion inhibited p-IκB levels and NF-κB activity and its overexpression increased p-IκB expression and NF-κB activity. NF-κB inhibitor BAY 11-7082 reversed the role of CARMA3 on cyclin D1 upregulation. In conclusion, our study found that CARMA3 is overexpressed in colon cancers and contributes to malignant cell growth by facilitating cell cycle progression through NF-κB mediated upregulation of cyclin D1.     

非小细胞肺癌中NF-κ B、VEGF 的表达及其与肿瘤血管形成的研究

目的:探讨核因子-κB(NF-κB)和血管内皮生长因子(VEGF)在非小细胞肺癌中的表达及其与肿瘤血管形成的关系。方法:应用免疫组织化学方法检测56例非小细胞肺癌及20例癌旁肺组织中的NF-κB P65、VEGF的表达,并用抗CD34测定肿瘤血管的密度(MVD)。结果:(1)在非小细胞肺癌中NF-κB P65、VEGF的表达阳性率分别为83.9%(47/56)、69.6%(39/56),明显高于癌旁组织(P<0.05);(2)NF-κB P65的表达在不同的TNM分期、淋巴结及胸腔积液、吸烟的分组之间差异有统计学意义,VEGF的表达在淋巴结及胸膜转移之间差异有统计学意义;(3)NF-κB P65、VEGF、MVD三者间存在明显相关性。结论:NF-κB、VEGF异常表达与NSCLC的发生、发展及肿瘤血管的形成密切的关系。     

NF—κB、VEGF在非小细胞肺癌中的表达及其与肿瘤血管形成的研究

目的探讨核因子-κB（NF-κB）和血管内皮生长因子（VEGF）在非小细胞肺癌（NSCLC）中的表达及其与肿瘤血管形成的关系。方法应用免疫组织化学方法,检测56例非小细胞肺癌及20例癌旁肺组织中的NF-κBP65、VEGF的表达,并用抗CD34测定肿瘤血管的密度（MVD）。结果（1）在非小细胞肺癌中NF-κBP65、VEGF的表达阳性率分别为83．9％（47／56）、69．6％（39／56）,明显高于癌旁组织（P〈O．05）;（2）NF-κBP65的表达在不同的TNM分期、淋巴结及胸腔积液、吸烟的分组之间差异有统计学意义,VEGF的表达在淋巴结及胸膜转移之间差异有统计学意义;（3）NF-κBP65、VEGF、MVD三者间存在明显相关性。结论NF-κB、VEGF异常表达与NSCLC的发生、发展及肿瘤血管的形成有着密切的关系。     

NF-κB、IκB、IKK在非小细胞肺癌中的表达及意义

目的检测非小细胞肺癌(non-small cell lung cancer,NSCLC)中NF-κB P65、p-IκBα(IκBα磷酸化)、p-IKKβ(IKKβ磷酸化)的表达情况及其与NSCLC临床特征的关系。方法采用免疫组化Elivision法检测NF-κB P65、p-IκBα、p-KKβ在56例NSCLC中表达情况,以20例癌旁组织作为对照。结果在NSCLC中NF-κB P65、p-IκBα、p-IKKβ的表达阳性率分别为83.9%(47/56)、55.7%(31/56)、69.6%(39/56),癌旁组织三者分别为20%(4/20)、25%(5/20)、30%(6/20),NF-κB P65、p-IκBα、p-IKKβ的表达与吸烟史、TNM分期、淋巴结转移相关,差异有统计学意义(P<0.05)。结论 NF-κB P65、p-IκBα、p-IKKβ高表达与NSCLC的发生、发展起着重要作用。     

CARMA3: A novel scaffold protein in regulation of NF-κB activation and diseases

CARD recruited membrane associated protein 3 (CARMA3) is a novel scaffold protein. It belongs to the CARMA protein family, and is known to activate nuclear factor (NF)-κB. However, it is still unknown which receptor functions upstream of CARMA3 to trigger NF-κB activation. Recently, several studies have demonstrated that CARMA3 serves as an indispensable adaptor protein in NF-κB signaling under some G protein-coupled receptors (GPCRs), such as lysophosphatidic acid (LPA) receptor and angiotensin (Ang) II receptor. Mechanistically, CARMA3 recruits its essential downstream molecules Bcl10 and MALT1 to form the CBM (CARMA3-Bcl10-MALT1) signalosome whereby it triggers NF-κB activation. GPCRs and NF-κB play pivotal roles in the regulation of various cellular functions, therefore, aberrant regulation of the GPCR/NF-κB signaling axis leads to the development of many types of diseases, such as cancer and atherogenesis. Recently, the GPCR/CARMA3/NF-κB signaling axis has been confirmed in these specific diseases and it plays crucial roles in the pathogenesis of disease progression. In ovarian cancer cell lines, knockdown of CARMA3 abolishes LPA receptor-induced NF-κB activation, and reduces LPA-induced ovarian cancer invasion. In vascular smooth cells, downregulation of CARMA3 substantially impairs Ang-II-receptor-induced NF-κB activation, and in vivo studies have confirmed that Bcl10-deficient mice are protected from developing Ang-II-receptor-induced atherosclerosis and aortic aneurysms. In this review, we summarize the biology of CARMA3, describe the role of the GPCR/CARMA3/NF-κB signaling axis in ovarian cancer and atherogenesis, and speculate about the potential roles of this signaling axis in other types of cancer and diseases. With a significant increase in the identification of LPA- and Ang-II-like ligands, such as endothelin-1, which also activates NF-κB via CARMA3 and contributes to the development of many diseases, CARMA3 is emerging as a novel therapeutic target for various types of cancer and other diseases.     

Id-1 promotes migration and invasion of non-small cell lung cancer cells through activating NF-κB signaling pathway

Background

Numerous studies have shown that Id-1 (Inhibitor of differentiation 1) is upregulated in several cancers and associated with tumor malignant characters. However, the clinical significance and biological role of Id-1 in non-small cell lung cancer (NSCLC) remains unclear.

Methods

We used RT-PCR, Western blot and Immunohistochemistry to measure Id-1 expression in NSCLC tissues and matched adjacent noncancerous tissues. The expression pattern of Id-1 in NSCLC tissues was determined by scoring system of immunohistochemical analysis. The Kaplan-Meier method was used to calculate the survival curve, and log-rank test to determine statistical significance. The Id-1 gene was overexpressed or downreuglated with Lentiviral vectors in NSCLC cells. And, the migration ability of NSCLC cells was tested in a Transwell Boyden Chamber.

Results

We found that Id-1 is generally expressed higher in NSCLC tissues compared with matched adjacent noncancerous tissues. We also found that high Id-1 expression in tumor tissues is significantly correlated with tumor progression and poor survival in NSCLC patients. Furthermore, our experimental data revealed that knockdown of Id-1 significantly suppressed the proliferation, migration and invasion of NSCLC cells, whereas ectopic expression of Id-1 promoted the malignant phenotype of NSCLC cells. Mechanistic study showed that NF-κB signaling pathway contributed to the effects of Id-1 in NSCLC cells. Moreover, blocking the NF-κB pathway significantly inhibited the tumor-promoting actions of Id-1 in NSCLC cells.

Conclusions

We identified a tumorigenic role of Id-1 in NSCLC and provided a novel therapeutic target for NSCLC patients.

     

Fn14 在 EGFR 外显子 19 缺失突变的非小细胞肺癌中的表达及其临床意义

目的:探讨成纤维细胞生长因子诱导14(Fn14)在表皮生长因子受体(EGFR)外显子19缺失的非小细胞肺癌(NSCLC)组织中的表达及其临床意义。方法:选择2010年9月至2013年11月第四军医大学唐都医院收治的125例原发性EGFR外显子19缺失的NSCLC组织及与之相对应的30例正常肺组织为研究对象,应用免疫组化法检测和比较其Fn14的表达,分析Fn14的表达与原发性EGFR外显子19缺失的的NSCLC临床病理特征的关系。结果:Fn14阳性表达主要定位于胞膜和胞质中。在125例原发性EGFR外显子19缺失的NSCLC组织中,Fn14的阳性表达率为100%,显著高于正常肺组织(P<0.001)。鳞癌和腺癌NSCLC组织中Fn14的阳性表达率比较无统计学差异(P=0.106);不同病理分化程度、不同TNM分期的NSCLC组织中Fn14的阳性表达率比较均具有显著性差异(P=0.000、P=0.000)。Fn14蛋白的表达与EGFR外显子19缺失的NSCLC的淋巴结转移状态、肿瘤大小以及肿瘤解剖学分类均显著相关(P=0.000、P=0.029、P=0.000、P=0.026),而与患者的性别、年龄、吸烟史均无关(P=0.816、P=0.122、P=0.816)。结论:Fn14在原发性EGFR外显子19缺失的NSCLC中呈异常高表达,EGFR外显子19缺失突变很可能通过上调Fn14通路,促进NSCLC的发生和发展。     

Adenosine triphosphate regulates NADPH oxidase activity leading to hydrogen peroxide production and COX-2/PGE2 expression in A549 cells

Non-small cell lung carcinoma (NSCLC) accounts for most of all lung cancers, which is the leading cause of mortality in human beings. High level of cyclooxygenase-2 (COX-2) is one of the features of NSCLC and related to the low survival rate of NSCLC. However, whether extracellular nucleotides releasing from stressed resident tissues contributes to the expression of COX-2 remains unclear. Here, we showed that stimulation of A549 cells by adenosine 5'-O-(3-thiotriphosphate) (ATPγS) led to an increase in COX-2 gene expression and prostaglandin E(2) (PGE(2)) synthesis, revealed by Western blotting, RT-PCR, promoter assay, and enzyme-linked immunosorbent assay. In addition, ATPγS induced intracellular reactive oxygen species (ROS) generation through the activation of NADPH oxidase. The increase of ROS level resulted in activation of the c-Src/epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor (NF)-κB cascade. We also found that activated Akt was translocated into the nucleus and recruited with NF-κB and p300 to form a complex. Thus, activation of p300 modulated the acetylation of histone H4 via the NADPH oxidase/c-Src/EGFR/PI3K/Akt/NF-κB cascade stimulated by ATPγS. Our results are the first to show a novel role of NADPH oxidase-dependent Akt/p65/p300 complex formation that plays a key role in regulating COX-2/PGE(2) expression in ATPγS-treated A549 cells. Taken together, we demonstrated that ATPγS stimulated activation of NADPH oxidase, resulting in generation of ROS, which then activated the downstream c-Src/EGFR/PI3K/Akt/NF-κB/p300 cascade to regulate the expression of COX-2 and synthesis of PGE(2) in A549 cells. Understanding the regulation of COX-2 expression and PGE(2) release by ATPγS on A549 cells may provide potential therapeutic targets of NSCLC.     

Correlation of epithelial-mesenchymal transition markers with clinicopathologic parameters in adenocarcinomas and squamous cell carcinoma of the lung

Epithelial-mesenchymal transition (EMT) is characterized by the loss of epithelial cell junction proteins and the gain of mesenchymal markers. The aim of this study was to analyze the associations between the EMT-related markers vimentin, E-cadherin, β-catenin, slug, snail, and twist1 and clinicopathologic parameters as well as epidermal growth factor receptor (EGFR) gene copy number and protein expression in non-small cell lung carcinoma (NSCLC). Fifty-nine squamous cell carcinomas (SCC) and 43 adenocarcinomas (AD) were immunohistochemically examined for respective EMT markers and for EGFR, using the EGFR PharmDx kit (Dako) for protein expression and automated silver enhanced in situ hybridization (SISH) for copy number. Vimentin expression in tumor epithelia was significantly higher in AD samples than in SCC samples (P=0.015). Among AD samples, vimentin expression was positively correlated with histologic grade (2 vs. 3; P=0.021) and exhibited a tendency toward a positive correlation with pTNM stage (I vs. II-IV; P=0.052). EGFR gene copy number was positively correlated with EGFR protein expression among both AD samples (P=0.008) and SCC samples (P=0.042), with EGFR protein expression being significantly higher in SCC samples compared with AD (P=0.038). Among AD samples, EGFR protein expression was associated with higher cytoplasmic expression of β-catenin (P=0.031). Among SCC samples, EGFR protein expression was negatively correlated with nuclear expression of β-catenin (P=0.033) but positively with nuclear slug (P=0.021). The expression pattern of EMT markers in AD suggests that vimentin is a possible immunohistochemical predictor of tumor progression.     

Angio-associated migratory cell protein interacts with epidermal growth factor receptor and enhances proliferation and drug resistance in human non-small cell lung cancer cells

Angio-associated migratory cell protein (AAMP) is expressed in some human cancer cells. Previous studies have shown AAMP high expression predicted poor prognosis. But its biological role in non-small cell lung cancer (NSCLC) cells is still unknown. In our present study, we attempted to explore the functions of AAMP in NSCLC cells. According to our findings, AAMP knockdown inhibited lung cancer cell proliferation and inhibited lung cancer cell tumorigenesis in the mouse xenograft model. Epidermal growth factor receptor (EGFR) is a primary receptor tyrosine kinase (RTK) that promotes proliferation and plays an important role in cancer pathology. We found AAMP interacted with EGFR and enhanced its dimerization and phosphorylation at tyrosine 1173 which activated ERK1/2 in NSCLC cells. In addition, we showed AAMP conferred the lung cancer cells resistance to chemotherapeutic agents such as icotinib and doxorubicin. Taken together, our data indicate that loss of AAMP from NSCLC inhibits tumor growth and elevates drug sensitivity, and these findings have clinical implications to treat NSCLC cancers.     

Down-regulation of epidermal growth factor receptor by curcumin-induced UBE1L in human bronchial epithelial cells

UBE1L, ubiquitin-activating enzyme E1-like, is the activating enzyme of ISG15ylation (ISG15, interferon stimulated gene 15). Loss of UBE1L and activation of epidermal growth factor receptor (EGFR) signaling are common events in lung carcinogenesis. Curcumin, a well-studied chemopreventive agent, is known to down-regulate EGFR. The present study demonstrated that curcumin decreased EGFR expression in human bronchial epithelial (HBE) Beas-2B cells and lung cancer A549 cells. For the first time, UBE1L was found to be induced by curcumin in HBE cells. Interestingly, overexpression of UBE1L reduced EGFR at posttranslational level in HBE cells. UBE1L triggered EGFR membrane internalization and promoted complex formation between ISG15 and EGFR. Curcumin decreased EGFR downstream signaling pAKT and nuclear factor κB (NF-κB). Overexpression or knockdown of UBE1L also resulted in down-regulation or up-regulation of phosphoinositide 3-kinase/AKT/NF-κB correspondently. In human samples, there was an inverse relationship between UBE1L and EGFR/AKT/NF-κB in non-small cell lung cancer tissues and adjacent tissues. These results uncover a novel chemopreventive mechanism of curcumin in inducing UBE1L and down-regulating EGFR signaling in HBE cells.     

非小细胞肺癌EGFR表达与^18F-FDG-PET SUVmax值的相关性研究

目的：探讨非小细胞肺癌组织中EGFR蛋白在的表达与PET-CT检测的SUVmax的相关性。方法：临床收集30例非小细胞肺癌患者,每一病例首先行PET-CT检查,然后行CT导引下活检穿刺,同一病灶取2个穿刺部位,分别为肿瘤组织内SUVmax值2.5-5区域及SUVmax〉5区域,采用免疫组化方法检测检测30例非小细胞肺癌肿瘤内部不同部位EGFR表达。结果：EGFR蛋白表达、SUVmax值在不同年龄、性别、病理类型中表达的差异无统计学意义（P均〉0.05）。EGFR蛋白表达、SUVmax值在不同的肿瘤直径、临床分期组表达差异有统计学意义（P均〈0.05）。NSCLC肿块SUVmax值与组织EGFR表达呈显著正相关（P〈0.05）。结论：NSCLC组织SUV值与组织EGFR表达有显著相关性,可以为非小细胞肺癌的放疗提供指导。     

非小细胞肺癌靶向治疗的临床前研究进展

针对表皮生长因子受体（EGFR）和血管生成（angiogenesis）信号通路的靶向治疗已经在晚期非小细胞肺癌的治疗上取得成功,但由于抗药性的存在,大多数晚期患者的生存时间仍然提高有限。继发性的EGFR T790M突变和原癌基因肝细胞生长因子受体（MET）的扩增被鉴定为两种主要的抗药机制。最近转化生长因子-β（TGF-β）/白介素-6信号通路被报道能介导选择性和适应性地对erlotinib的抗药。另一方面,Kras突变所致肺癌的靶向治疗方面也取得了一些进展。双重抑制磷脂酰肌醇3-激酶（PI3K）和促分裂素原活化蛋白激酶激酶（MEK）信号通路可导致Kras突变肿瘤的显著消退,联合抑制SRC、PI3K和MEK可使丝氨酸/苏氨酸蛋白激酶11（Lkb1）缺失,Kras突变的肺癌小鼠的肿瘤明显消退,抑制核因子-κB（NF-κB）信号通路导致p53缺失,Kras突变的肿瘤发展显著减慢。这些发现都为发展非小细胞肺癌患者的靶向治疗提供了有力的支持。     

Protein kinase C-δ negatively regulates T cell receptor-induced NF-κB activation by inhibiting the assembly of CARMA1 signalosome

T-cell receptor (TCR)-induced T-cell activation is a critical event in adaptive immune responses. The engagement of TCR complex by antigen along with the activation of the costimulatory receptors trigger a cascade of intracellular signaling, in which caspase recruitment domain-containing membrane-associated guanylate kinase 1 (CARMA1) is a crucial scaffold protein. Upon stimulation, CARMA1 recruits downstream molecules including B-cell CLL/lymphoma 10 (Bcl10), mucosa-associated lymphoid tissue lymphoma translocation gene 1 (MALT1), and TRAF6 to assemble a specific TCR-induced signalosome that triggers NF-κB and JNK activation. In this report, we identified protein kinase Cδ (PKCδ) as a CARMA1-associated protein by a biochemical affinity purification approach. PKCδ interacted with CARMA1 in TCR stimulation-dependent manner in Jurkat T cells. Overexpression of PKCδ inhibited CARMA1-mediated NF-κB activation, whereas knockdown of PKCδ potentiated TCR-triggered NF-κB activation and IL-2 secretion in Jurkat T cells. Reconstitution experiments with PKCδ kinase-dead mutant indicated that the kinase activity of PKCδ was dispensable for its ability to inhibit TCR-triggered NF-κB activation. Furthermore, we found that PKCδ inhibited the interaction between MALT1 and TRAF6, but not the association of CARMA1 with PKCθ, Bcl10, or MALT1. These observations suggest that PKCδ is a negative regulator in T cell activation through inhibiting the assembly of CARMA1 signalosome.     

Methylation of TMEFF2 gene in tissue and serum DNA from patients with non-small cell lung cancer

Lung cancer remains a global health problem with a high mortality rate. CpG island methylation is a common aberration frequently associated with gene silencing in multiple tumor types, emerging as a highly promising biomarker. The transmembrane protein with a single EGF-like and two follistatin domains (TMEFF2) is epigenetically silenced in numerous tumor types, suggesting a potential role as a potential tumor suppressor. However, the role of TMEFF2 in lung cancer remains to be fully elucidated. We explored the methylation status of TMEFF2 gene in 139 patients with non-small cell lung cancer (NSCLC) and the feasibility of detecting circulating methylated DNA as a screening tool for NSCLC using methylation-specific PCR in 316 patients and 50 age-matched health controls. TMEFF2 methylation in tumor tissues was found in 73 of the 139 NSCLCs (52.5%) and was related to gene expression. The frequency of TMEFF2 methylation was higher in females and never-smokers than in males and smokers with borderline significance (65.8% vs 47.8%, p = 0.06; 65.7% vs 48.1%, p = 0.07). Notably, in adenocarcinomas, TMEFF2 methylation was significantly more frequent in tumors without EGFR mutation than those with EGFR mutation (adjusted odds ratio = 7.13, 95% confidence interval = 2.05-24.83, P = 0.002). Furthermore, TMEFF2 methylation was exclusively detected in the serum of NSCLC patients at a frequency of 9.2% (29/316). These findings suggest that methylation-associated down-regulation of TMEFF2 gene may be involved in lung tumorigenesis and TMEFF2 methylation can serve as a specific blood-based biomarker for NSCLC.     

非小细胞肺癌EGFR 表达与18F-FDG-PET SUVmax 值 的相关性研究

目的:探讨非小细胞肺癌组织中EGFR蛋白在的表达与PET-CT检测的SUVmax的相关性。方法:临床收集30例非小细胞肺癌患者,每一病例首先行PET-CT检查,然后行CT导引下活检穿刺,同一病灶取2个穿刺部位,分别为肿瘤组织内SUVmax值2.5-5区域及SUVmax>5区域,采用免疫组化方法检测检测30例非小细胞肺癌肿瘤内部不同部位EGFR表达。结果:EGFR蛋白表达、SUVmax值在不同年龄、性别、病理类型中表达的差异无统计学意义(P均>0.05)。EGFR蛋白表达、SUVmax值在不同的肿瘤直径、临床分期组表达差异有统计学意义(P均<0.05)。NSCLC肿块SUVmax值与组织EGFR表达呈显著正相关(P<0.05)。结论:NSCLC组织SUV值与组织EGFR表达有显著相关性,可以为非小细胞肺癌的放疗提供指导。     

NF-kB 与Slug 在非小细胞肺癌及其上皮间质转化中的作用

目的:研究核因子NF-kB与slug在非小细胞肺癌(NSCLC)中的表达情况、及二者与非小细胞肺癌上皮间质转化(EMT)的关系,为非小细胞肺癌的诊断治疗提供理论依据。方法:(1)采用免疫组化PV9000二步法测定50例NSCLC组织及20例相应正常肺组织中NF-kBP65、slug、E-cadherin及Vimentin蛋白表达情况。(2)采用RT-PCR测定其中25例NSCLC组织及10例相应正常肺组织中NF-kBP65、slug的mRNA表达情况。结果:NSCLC中NF-kBP65蛋白表达量高于癌旁正常肺组织(Z=-2.370,P<0.05),NF-kBP65mRNA表达量明显高于癌旁正常肺组织(t=4.967,P<0.01);Slug蛋白表达量明显高于癌旁正常肺组织(Z=-4.443,P<0.01),SlugmRNA表达量明显高于癌旁正常肺组织(t=6.483,P<0.01)。在NF-kBP65阳性癌组织中,E-cadherin蛋白表达下降(x2=5.024,P<0.05),Vimentin蛋白表达上升(x2=4.723,P<0.05);Slug阳性癌组织中,E-cadherin蛋白表达下调(x2=5.984,P<0.05),Vimentin表达上调(x2=5.028,P<0.05)。另外,NF-kBP65与Slug在蛋白水平呈极显著正相关(r=0.443,P<0.01),在mRNA水平呈显著正相关(r=0.439,P<0.05)。NF-kB与分化程度(x2=5.024,P<0.05)、有无淋巴结转移(x2=7.933,P<0.01)及肿瘤的分期(x2=5.614,P<0.05)有关,与性别、年龄、组织类型无明显相关性(P>0.05);Slug与淋巴结转移(x2=6.174,P<0.05)及肿瘤的分期(x2=7.317,P<0.01)有关,与性别、年龄、组织类型、分化程度无明显相关性(P>0.05)。结论:NF-kB、Slug在NSCLC中表达增强,可能与NSCLC的发生、发展、转移有关;并且NF-kB与Slug可能协同抑制E-cadherin表达,促进Vimentin表达,诱使NSCLC的EMT发生,从而为进一步研究NSCLC的EMT提供理论依据。     

Thrombin-dependent NF-{kappa}B activation and monocyte/endothelial adhesion are mediated by the CARMA3·Bcl10·MALT1 signalosome

Thrombin is a potent modulator of endothelial function and, through stimulation of NF-κB, induces endothelial expression of intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). These cell surface adhesion molecules recruit inflammatory cells to the vessel wall and thereby participate in the development of atherosclerosis, which is increasingly recognized as an inflammatory condition. The principal receptor for thrombin on endothelial cells is protease-activated receptor-1 (PAR-1), a member of the G protein-coupled receptor superfamily. Although it is known that PAR-1 signaling to NF-κB depends on initial PKC activation, the subsequent steps leading to stimulation of the canonical NF-κB machinery have remained unclear. Here, we demonstrate that a complex of proteins containing CARMA3, Bcl10, and MALT1 links PAR-1 activation to stimulation of the IκB kinase complex. IκB kinase in turn phosphorylates IκB, leading to its degradation and the release of active NF-κB. Further, we find that although this CARMA3·Bcl10·MALT1 signalosome shares features with a CARMA1-containing signalosome found in lymphocytes, there are significant differences in how the signalosomes communicate with their cognate receptors. Specifically, whereas the CARMA1-containing lymphocyte complex relies on 3-phosphoinositide-dependent protein kinase 1 for assembly and activation, the CARMA3-containing endothelial signalosome functions completely independent of 3-phosphoinositide-dependent protein kinase 1 and instead relies on β-arrestin 2 for assembly. Finally, we show that thrombin-dependent adhesion of monocytes to endothelial cells requires an intact endothelial CARMA3·Bcl10·MALT1 signalosome, underscoring the importance of the signalosome in mediating one of the most significant pro-atherogenic effects of thrombin.     

nm23、EGFR和Bcl-2蛋白在肺癌组织中的表达及临床意义

目的:观察nm23、EGFR和Bcl-2在人肺癌中的表达,探讨其与肺癌临床生物学行为的关系。方法:采用免疫组化检测了49例肺癌及癌旁组织中nm23、EGFR和Bcl-2蛋白的表达情况,并对其与肺癌临床生物学行为进行相关分析。结果:在49例肺癌中,EGFR和Bel-2蛋白的阳性表达率分别为51.0%和34.7%,均高于正常肺组织17.5%和12.5%(p<0.05),nm23蛋白的阳性表达率为55.1%,则显著低于正常肺组织87.5%(p<0.01)。在病理分级的高一中分化、低分化和未分化中nm23的阳性表达率分别为82.6%、41.2%和11.1%(p<0.01)。EGFR在非小细胞肺癌(NSCLC)组为61.5%,在小细胞肺癌(SCLC)组为10.0%(p<0.01)。在NSCLC TNM分期中,Ⅰ期 Ⅱ期与Ⅲ P<0.01)期中EGFR的阳性表达率分别为39.3%和81.8%(p<0.01);病理分级的高-中分化和低分化中EGFR的阳性表达率分别为39.1%和88.2%(p<0.01);在淋巴结转移组为57.9%,无淋巴结转移组为27.3%(p<0.05)。Bcl-2的阳性表达率在NSCLC组为25.6%,在SCLC组为70%,统计学分析差异有统计学意义(p<0.01);在NSCLC组病理分级的高分化、中分化和低分化中Bcl-2阳性表达率分别为62.5%、26.7%和5.89%(p<0.01)。结论:癌基因EGFR、凋亡抑制基因Bcl-2与抑癌基因nm23在肺癌的发生、发展中存在着相互促进作用。