Expression of Bcl-2 and Amplification of c-myc Are Frequent in Basaloid Squamous Cell Carcinomas of the Esophagus

Basaloid squamous cell carcinoma (BSCC) of the esophagus is a rare, poorly differentiated variant of typical esophageal squamous cell carcinoma (SCC) characterized by high proliferative activity and frequent spontaneous apoptoses. In the present study, we investigated the expression of the apoptosis-suppressing protein Bcl-2 in 23 BSCC of the esophagus and 23 stage-matched typical esophageal SCC by means of immunohistochemistry. In addition, amplification of the apoptosis- and proliferation-inducing gene c-myc was determined by means of differential polymerase chain reaction. Bcl-2 expression was found significantly more often in BSCC than in SCC (86.9% vs. 17.4%, P < 0.0001). Amplification of c-myc was nearly twice as common in BSCC as in SCC (47.8% vs. 26.1%, not significant). Bcl-2 protein expression together with c-myc amplification was detected in 43.5% of the BSCC but in none of the typical SCC (P < 0.0001). Taken together, our findings indicate that the molecular pathogenesis of esophageal BSCC differs from that of typical SCC and frequently involves coactivation of c-myc and Bcl-2.     

喉癌组织中MKRN1和HSP90基因表达的研究

【摘要】目的 正常细胞中MKRN1和HSP90基因的平衡决定了端粒酶活性和端粒长度,使细胞保持正常的状态。二者的表达异常可能与肿瘤的发生发展有关。为探讨MKRN1和HSP90基因的表达与喉癌发生发展之间的关系,我们研究了喉癌组织中MKRN1和HSP90基因mRNA的表达情况。方法 提取35例喉癌组织的总mRNA,用半定量RT-PCR的方法检测MKRN1和HSP90基因mRNA的表达情况。结果 35例喉鳞癌中有18例MKRN1基因mRNA表达下调,占51.4%（18/35）;16例HSP90基因mRNA表达上调,占45.7%（16/35）。 MKRN1基因在肿瘤组织中的表达水平为0.74±0.63,在癌旁正常对照组织中为1.21±0.76,（P＜0.01）。HSP90基因在肿瘤组织中表达水平为1.82±1.25,而在癌旁正常对照组织中为1.23±1.04,（P＜0.05）,统计学上具有显著差异。MKRN1与HSP90基因的表达存在负相关(r=-0.458, P＜0.05)。HSP90基因表达情况与肿瘤的病理分期和患者年龄相关 (P＜0.05) ,MKRN1的表达与肿瘤的临床特征均无相关性(P＞0.05)。结论 喉癌组织中MKRN1和HSP90基因mRNA的表达异常存在相关性,二者表达平衡的破坏可能在喉癌的发生发展中起着重要的作用。     

口腔鳞癌组织中P15和P21的表达研究

目的探讨p15和p21基因的蛋白产物在口腔鳞癌中的表达及意义。方法采用免疫组化P—V法检测108例口腔鳞癌和10例正常口腔黏膜组织中P15和P21蛋白的表达，并将结果进行统计分析。结果口腔鳞癌组织中P15蛋白的阳性表达率低于正常口腔黏膜组织（P〈0．05），而P21蛋白的阳性表达率两组间无明显差异（P〉0．05）；口腔鳞癌中，Ⅲ、Ⅳ期口腔鳞癌P15蛋白的阳性表达率明显低于Ⅰ、Ⅱ期口腔鳞癌的表达率（P〈0．01），无颈淋巴结转移组P15蛋白表达水平比有转移组高（P〈0．05）；P21阳性表达率在不同性别组中的表达差异具有统计学意义（P〈0．05）。结论P15蛋白的表达与口腔鳞癌的临床分期及淋巴结转移有关，P15蛋白表达水平可作为临床评估口腔鳞癌恶性程度、转移和预后的参考指标。     

TRF2在食管鳞癌中的表达及意义

目的：探讨端粒重复序列结合因子2（Telomeric repeat binding factor2,TRF2）在食管病变中的表达及其临床意义。方法：采用免疫组化ABC法检测60例食管鳞状细胞癌、30例食管上皮内瘤变和20例食管正常组织中TRF2的表达,并探讨与食管癌临床病理特征之间的关系。结果：在正常上皮组织、上皮内瘤变和食管鳞癌组织中,TRF2的阳性表达率分别为35%、56.7%和96.7%。食管鳞癌组织中,TRF2的表达与淋巴结转移和病理分级呈轻度正相关（P〈0.01）。结论：TRF2在食管癌组织中表达异常增高,提示TRF2与食管癌的发生、发展有密切的关系,检测TRF2的表达可能成为判断食管鳞癌生物学行为的重要指标。     

Adhesion Protein ApfA of Actinobacillus pleuropneumoniae Is Required for Pathogenesis and Is a Potential Target for Vaccine Development

Actinobacillus pleuropneumoniae is the etiologic agent of porcine pleuropneumonia, which causes serious economic losses in the pig farming industry worldwide. Due to a lack of knowledge of its virulence factors and a lack of effective vaccines able to confer cross-serotype protection, it is difficult to place this disease under control. By analyzing its genome sequences, we found that type IV fimbrial subunit protein ApfA is highly conserved among different serotypes of A. pleuropneumoniae. Our study shows that ApfA is an adhesin since its expression was greatly upregulated (135-fold) upon contact with host cells, while its deletion mutant attenuated its capability of adhesion. The inactivation of apfA dramatically reduced the ability of A. pleuropneumoniae to colonize mouse lung, suggesting that apfA is a virulence factor. Purified recombinant ApfA elicited an elevated humoral immune response and conferred robust protection against challenges with A. pleuropneumoniae serovar 1 strain 4074 and serovar 7 strain WF83 in mice. Importantly, the anti-ApfA serum conferred significant protection against both serovar 1 and serovar 7 in mice. These studies indicate that ApfA promotes virulence through attachment to host cells, and its immunogenicity renders it a promising novel subunit vaccine candidate against infection with A. pleuropneumoniae.     

Platelet-Derived Growth Factor-B Normalizes Micromorphology and Vessel Function in Vascular Endothelial Growth Factor-A-Induced Squamous Cell Carcinomas

Vascular endothelial growth factor (VEGF), which is a key regulator of angiogenesis, often induces formation of immature vessels with increased permeability and reduced vessel functionality. Here, we demonstrate that de novo expression of murine (m)VEGF-164 induces malignant and invasive tumor growth of HaCaT keratinocytes. However, the mVEGF-164-induced tumors are ulcerated with a disorganized epithelium that is interrupted by lacunae with limited basement membrane and endothelial cell coverage. Vessel maturation is strongly impaired. Tumor and vessel micromorphology are markedly improved by the combined expression of human platelet-derived growth factor (hPDGF)-B and mVEGF-164. Although tumor size and malignancy are comparable with either mVEGF-164 alone or combined human PDGF-B and mVEGF-164 expression, combined hPDGF-B and mVEGF-164 expression leads to a more solid and compact tumor tissue with a mature functional tumor vasculature and a higher microvessel density, as demonstrated histologically and by dynamic contrast-enhanced magnetic resonance imaging. Treatment of the hPDGF-B- and mVEGF-164-expressing tumors with imatinib mesylate to block PDGF-B signaling reverses this effect. In addition, tumor cell invasion of mVEGF-164 transfectants and mVEGF-164 plus hPDGF-B transfectants in vivo is associated with a marked induction of tumor-derived matrix metalloproteinase-1 and stromal matrix metalloproteinase-9 and -13, as was confirmed in three-dimensional organotypic co-cultures with fibroblasts in vitro. These data clearly demonstrate the need for a concerted action of different growth factors in the establishment of solid tumors with functional vasculature and emphasize the need for a multifactorial therapy.Tumor angiogenesis is a complex multistep process and a crucial pre-requisite for tumor development, growth and progression.¹ It includes the degradation of the extracellular matrix as well as the proliferation, migration, and differentiation of endothelial cells and is tightly controlled by stimulatory and inhibitory factors.^2,3,4 One of the key angiogenic factors is vascular endothelial growth factor (VEGF), also known as vascular permeability factor, which plays an important role in physiological and pathological angiogenesis, eg, during wound healing^5,6 and cancer.^7,8 VEGF is up-regulated by different stimuli, eg, hypoxia⁹ or ras oncogene activation,^10,11 and its overexpression in tumor cells enhances tumor growth^12,13,14 and invasion.^15,16,17 However, multiple studies have demonstrated that VEGF-induced blood vessels often show increased permeability, resulting in hemorrhages and reduced vessel functionality.¹⁸ These observations indicate that additional growth factors cooperate with VEGF during normal and pathological angiogenesis to establish mature, functional blood vessels with an intact basement membrane that can promote tumor growth by supplying the necessary nutrients to the tumor cell.The platelet-derived growth factor (PDGF) isoform PDGF-B critically contributes to vessel stabilization by recruiting pericytes and smooth muscle cells.^19,20 PDGF-B and PDGFR-β knockout mice failed to sufficiently recruit pericytes to newly formed blood vessels, leading to severe perturbations in blood vessel stabilization and maturation.^21,22 Lack of PDGF-B expression in endothelial cells resulted in the loss of pericytes around blood vessels and in vessel abnormalities.²³ Besides its functions in physiological angiogenesis, PDGF-B is up-regulated by many tumor cells,^24,25 where it can act as an autocrine growth factor. Yet it also plays a crucial role in the establishment of functional vessels in the tumor.^26,27 Tumor-inducing paracrine functions of PDGF-B have been demonstrated in our HaCaT skin carcinogenesis model. Transfection of the nontumorigenic keratinocyte cell line HaCaT with human (h)PDGF-B resulted in benign tumor growth that was mediated by initial stromal activation and subsequent stromal and vascular maturation.^28,29 In this context, we observed an early PDGF-B-mediated up-regulation of stromal VEGF that indicated a crucial role of VEGF in the induction of tumor growth.To further determine the specific role of VEGF in tumor growth, we stably transfected nontumorigenic HaCaT cells with murine (m)VEGF-A-164 and assessed tumor growth after s.c. injection and as surface transplants in nude mice. The combined effects of hPDGF-B and mVEGF-164 were analyzed using 1:1 mixed populations of hPDGF-B and mVEGF-164-transfected HaCaT cells in the s.c. tumor model and in the matrix inserted surface transplantation assay.We demonstrate that de novo expression of mVEGF-164 in the HaCaT cells induces malignant and invasive tumor growth. However, the mVEGF-164 transfectant tumors were ulcerated with a disorganized epithelium and interrupted by lacunae like structures with only limited basement membrane and endothelial cell coverage. In addition, blood vessel maturation was dramatically impaired. Tumor epithelial and vessel structures were markedly improved by the coexpression of hPDGF-B and mVEGF-164 in the 1:1 mixed cell populations, leading to a more compact tumor mass, an increased microvessel density and a mature functional tumor vasculature as demonstrated by histological analysis and by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI). The improved phenotype was reverted by blocking PDGF receptor (PDGFR) signaling with imatinib mesylate. These data clearly demonstrate that growth factors need to act in concert to establish solid tumor growth with a functional tumor vasculature.     

VAV3蛋白在喉鳞状细胞癌中的表达及作用机制

目的 研究VAV3蛋白在喉鳞状细胞癌组织中的表达情况,及其对喉癌临床分型、淋巴转移等影响,分析其可能的机制.方法 利用免疫组织化学方法检查VAV3蛋白在喉鳞状细胞癌组织及喉部非癌组织中的表达,检测VAV3蛋白对喉鳞状细胞癌侵袭,淋巴转移等影响.结果 VAV3蛋白在喉鳞状细胞癌组织中表达为69.76％,在喉部非癌组织中的表达为10％,前者明显高于后者,差异有统计学意义(x2=11.937,P=0.001).它的表达与临床分型、T分型及淋巴结转移有统计学意义(P＜0.05),与性别、年龄、吸烟、分型之间无统计学意义(P＞0.05).结论 VAV3蛋白在喉癌组织中高表达,且可促进喉癌组织侵袭、转移能力.VAV3蛋白高表达,可作为判断喉癌及预后的一种新的分子标志.     

The Tissue-Specific Stem Cell as a Target for Chemoprevention

While cancer treatment modalities are gradually improving due to increased knowledge about tumor heterogeneity and the cancer stem cell hypothesis, there remains a disconnect between tumor detection and mortality rates. The increasing knowledge of stem cell biology and its contribution to cancer progression illuminates the potential for chemopreventative regimens that effectively target the tissue-specific stem cell. Several signaling pathways have emerged that are critical for regulating stem cell self-renewal and multilineage differentiation over a range of tissue types, including Wnt, Hedgehog, and Notch signaling. Dysregulation of these genes can lead to cancer, which supports the cancer stem cell hypothesis. Several known chemopreventative agents have recently been shown to impact these and other pathways in the stem cell population, suggesting that their efficacies may be attributed in part to maintaining homeostasis of tissue-specific stem cells. Further understanding of the mechanisms of action of chemopreventative agents and of stem cell biology will generate better chemoprevention regimens that can be recommended especially to those in high-risk populations.     

A Role for the Epithelial Microenvironment at Tumor Boundaries : Evidence from Drosophila and Human Squamous Cell Carcinomas

Recent work has shown an increasing appreciation for the importance of the tumor environment, most commonly the overlying stroma. Less emphasis has been placed on the importance of local communication between transformed cells and their neighbors within the epithelium at tumor boundaries. We previously reported a Drosophila model that highlighted the importance of local interactions within the epithelial microenvironment: Src-transformed cells (Csk-deficient) were influenced by their immediate normal neighbors. The result was a consistent change in ‘border cells’ at the edge of transformed patches including delocalized p120-catenin and E-cadherin as well as invasive migration through the basal lamina. Here we show that the invasive properties of the boundary cells depend on up-regulation of Drosophila matrix metalloproteinase-1 as assessed by promoter activity, protein levels, in situ enzymatic activity, and tests of genetic modifier activity. Further, we provide evidence that these events at tumor borders may be evolutionarily conserved. We detected changes in ‘boundary cells’ within histological sections of human squamous cell carcinomas that were similar to those observed in Drosophila: both E-cadherin and p120-catenin exhibited normal junctional localization at the centers of the tumors but were reduced or delocalized at the boundary. Further, matrix metalloproteinase-2 was up regulated within these same boundary cells. These results support the view that local cell–cell interactions within the epithelial microenvironment impact tumor invasion and progression.Recent studies have highlighted the importance of interactions between tumors and their environment. Factors from the local microenvironment including extracellular matrix components, fibroblasts, and macrophages and other immune cells can influence the behavior of cancer cells both in terms of growth and metastasis.¹ Less understood is the influence of local interactions between transformed cells and their epithelial neighbors. These interactions are difficult to study because they occur early in tumor progression, and most human tumors are identified after tumors have significantly advanced. Recently, we presented data from a Drosophila tumor model that suggested the importance of early, local cell–cell interactions within the epithelium.² Here, we further explore this model and in addition offer evidence for this view from human oral squamous cell carcinomas (SCCs).The effects of the microenvironment are best studied in vivo. One system that offers the opportunity to study the effects of the microenvironment in situ is the fruit fly Drosophila. Fly models offer powerful genetic tools and a genome that contains orthologs of a majority of known oncogenes and tumor suppressors. As such, Drosophila is becoming increasingly popular as a cancer model (reviewed in 3, 4). Relevant to this article, the Drosophila tumor suppressor ortholog C-terminus Src Kinase (dCsk) was identified both as a factor mediating activity of the Lats/Warts tumor suppressor⁵ and oncogenic isoforms of the Ret receptor tyrosine kinase.⁶ Mammalian Csk is the major cellular inhibitor of the Src family kinases through direct phosphorylation.⁷ In Drosophila, broad loss of dCsk activity led to an increase in Src activity.^2,6We have previously reported effects from the epithelial microenvironment in cells deficient for dCsk.² When dCsk activity was reduced or removed within an entire epithelial region, dCsk cells exhibited ectopic growth—attributable to increased cell proliferation and decreased death—but retained epithelial integrity. By contrast, discrete dCsk-deficient patches of cells—bounded by wild-type cells—initiated a migratory and invasive program along the basal extracellular matrix. Such cellular behavior included (1) dissolution of the adherens junctions as assessed by delocalization of junction-associated proteins such as p120ctn, and (2) activation of the Rho GTPase and Jun kinase (JNK) pathway. The metalloproteinase dMmp2 was also required for this dCsk-dependent invasive migration phenotype.²In this study, we present evidence indicating that the remaining member of the Drosophila matrix metalloproteinase (MMP) family, dMmp1, is also required for the invasive phenotype of boundary dCsk cells. Our evidence includes genetic modifier interactions, promoter activation, increased protein levels, and in situ enzymatic activity. Remarkably, sections of human oral squamous cell carcinomas (OSCCs) exhibited a border difference strikingly similar to effects observed in our Drosophila model: human MMP2 was up-regulated at the borders of tumor islands whereas expression of the junctional markers E-cadherin and P120-catenin was decreased or delocalized in these same boundary cells. This indicates that the two models may share some aspects of tumor progression including the importance of local interactions within the epithelium.     

肿瘤标志物SURVIVIN及P27蛋白在肾细胞癌中表达的临床意义

目的：探讨新的凋亡相关基因Survivin蛋白和p27蛋白肿瘤标志物在人肾细胞癌中的表达及临床意义。方法：采用免疫组化SP法分别测定50例肾癌和12例正常肾组织中Survivin蛋白及p27蛋白的表达,并分析其与肾癌临床、病理的关系。结果：Survivin蛋白在肾癌组织中表达阳性率为68%（34/50）,而正常组织中未见表达,两者有统计学意义（P〈0.01）。肾癌中Survivin蛋白的表达与肾癌的病理分级呈正相关（P〈0.05）。而p27蛋白在正常肾组织中的阳性率为83.3%（10/12）,在肾癌组织中的阳性率为42%（21/50）,两者有统计学意义（P〈0.01）。p27蛋白表达与肾癌的分级呈负相关（P〈0.01）。结论：Survivin蛋白表达水平与肾癌的临床分期、淋巴结转移密切相关,且与p27蛋白呈负相关。联检Survivin蛋白和p27蛋白有助于提高对肾癌预后的评估,并可作为判定肾癌生物学行为的客观指标。     

肺癌骨转移患者99Tcm-MDP显像和鳞癌标志物对评估疗效的临床研究

目的 研究肺鳞癌患者化疗前后外周血CYFRA21-1和SCCA的表达水平以及与临床疗效的关系.方法 采用电化学发光法测定肺癌患者化疗前后血浆CYFRA21-1和SCCA含量.参照2009年实体瘤疗效评价标准RECIST,以SPECT99Tcm-MDP影像的病灶大小、数目在治疗前后的变化为评判依据,在化疗4周期后,疗效按完全缓解(CR)、部分缓解(PR)、稳定(SD)和进展(PD)分类.按临床获益(CB) =CR +PR,治疗无效(NR)=PD +SD分组.结果 CYFRA21-1水平在肺癌治疗无效组与临床获益组比较,差异有统计学意义(P＜0.01).SCCA水平在肺癌治疗无效组与临床获益组比较,差异有统计学意义(P＜0.01).治疗后SPECT病灶消失或缩小或数目减少的17例,无变化或有进展的23例.治疗后在CYFRA21-1水平高值组16例中,临床获益(CB)为18.75％,治疗无效(NR)为81.25％;在CYFRA21-1水平低值组24例中,CB为58.33％,NR为41.67％.两组比较,差异有统计学意义(x2=6.155,P＜0.05).治疗后在SCCA水平高值组14例中,CB为21.43％,NR为78.57％;在SCCA水平低值组26例中,CB为53.85％,NR为46.15％.两组比较,差异有统计学意义(x2=3.913,P＜0.05).结论 对比SPECT骨转移病灶大小、数目的变化,化疗前后血浆CYFRA21-1和SCCA水平变化对肺癌化疗效果评估具有较高的临床应用价值.与实体瘤疗效评价标准RECIST评估有一致性.     

Development of Keratoacanthomas and Squamous Cell Carcinomas in Transgenic Rabbits with Targeted Expression of EJras Oncogene in Epidermis

Activated ras genes have been frequently identified in both benign and malignant human tumors, including keratoacanthoma and squamous cell carcinoma. In this study, we developed two lines of transgenic rabbits in which the expression of EJras has been specifically targeted to the rabbit epidermal keratinocytes, using the upstream regulatory region of cottontail rabbit papillomavirus. All of the F1 transgenic progenies developed multiple keratoacanthomas at about 3 days after birth. The rabbits developed an average of 20 tumors, which usually reached the size of approximately 1 cm in diameter and then spontaneously regressed in about 2 months, similar to keratoacanthoma regression in humans. In addition, up to 18% of the rabbits then developed squamous cell carcinoma at about 5 months of age. The expression of EJras was detectable in all of the keratoacanthomas and squamous cell carcinomas. These results strongly support the involvement of the ras oncogene in both the initiation and regression of keratoacanthoma, and in the development of squamous cell carcinomas. These novel transgenic rabbits, with their consistent tumorigenic phenotype at an early age, high similarity to the human lesions, and easy accessibility for examination, manipulation, biopsy, and treatment, should provide a unique model system for studying ras activation-related tumor initiation, regression, and progression, and for evaluating antitumor therapies.     

Mitogen-Activated Protein Kinase Is Activated During IgG-Mediated Phagocytosis, But Is Not Required for Target Ingestion

Arachidonic acid (AA) release is required for IgG-mediated phagocytosis in human monocytes. AA release is mediated by a calcium-independent phospholipase A₂ (PPL) that is in turn regulated by protein kinase C (PKC). As mitogen-activated protein kinase (MAPK) activates cytosolic phospholipase A₂, we examined the activation and involvement of MAPK in IgG-mediated phagocytosis. MAPK activity was assessed in immunoprecipitates; tyrosine phosphorylation was detected by immunoblotting. Ingestion of IgG-opsonized glass beads, or treatment with phorbol myristate acetate, increased enzymatic activity and tyrosine phosphorylation of p42 MAPK. This MAPK activation was attenuated by PKC inhibitors staurosporine or calphostin C. Treatment with PD98059, a p42/p44 MAPK kinase (MEK) inhibitor, decreased BIgG-stimulated p42 MAPK activity by >90% with no significant effect on phagocytosis or pPL activity. These results suggest that p42 MAPK is activated in a PKC-dependent manner during IgG-dependent phagocytosis but is not required for target ingestion.     

The Expression of MicroRNA-155 in Plasma and Tissue Is Matched in Human Laryngeal Squamous Cell Carcinoma

PurposeTumor-associated microRNAs have been detected in cancer, though whether plasma microRNA-155 (miR-155) could be a potential biomarker for laryngeal squamous cell carcinoma (LSCC) prognosis is unclear. We aimed to determine how miR-155 can be used to predict the clinical characteristics of patients with LSCC and correctly diagnose them.ResultsA total of 280 LSCC patients and 560 age- and sex-matched controls were included in the study. The miR-155 level was more up-regulated in LSCC tissue than in the non-tumor tissues (13.6±2.4 vs. 3.1±0.80, p<0.001). Additionally, a significantly higher miR-155 level in plasma samples from LSCC patients than in those of the controls (8.9±1.25 vs. 1.8±0.8, p<0.001) was reported. Tissue miR-155 showed an area under the curve (AUC) of 0.933, with a sensitivity of 82.6% and a specificity of 89.2%. The AUC for plasma miR-155 was 0.757, with a sensitivity of 58.4% and a specificity of 69.5%. When early LSCC in TNM I stage was considered, tissue miR-155 showed an area under the curve of 0.804, with a sensitivity of 85.2% and a specificity of 87.3%.ConclusionThe expression of tissue and plasma miR-155 were significantly up-regulated in patients with LSCC. Our work will serve as a basis for further investigation, preferably large-scale validation in clinical trials.     

Involvement of CYR61 and CTGF in the Fascin-Mediated Proliferation and Invasiveness of Esophageal Squamous Cell Carcinomas Cells

Fascin is over-expressed in esophageal squamous cell carcinoma (ESCC) and involved in the proliferation and invasiveness of ESCC cells. In this study, we retrospectively examined the expression of fascin in ESCC samples by immunohistochemistry and revealed that overexpression of fascin was related to poor patient survival. RNAi-mediated knockdown of fascin in ESCC cells significantly inhibited cell proliferation and invasiveness, whereas forced expression of fascin in immortalized esophageal epithelial cells accelerated cell proliferation and invasiveness. To explore the underlying mechanism, cDNA microarray was performed to identify the differential gene expression profiles between a fascin-depleted cell line by RNAi and the corresponding control ESCC cells. Results showed that 296 genes were differentially expressed on fascin depletion. In this study, we focused on two down-regulated genes: CYR61 and CTGF. We found that restored expression of either CYR61 or CTGF led to a recovery of the suppression of cellular proliferation and invasiveness induced by down-regulation of fascin expression; the protein level of CYR61 and CTGF were up-regulated in ESCCs and their expression pattern correlated with fascin overexpression. Finally, analysis of signal transduction revealed that fascin affected the expressions of CYR61 and CTGF through transforming growth factor (TGF)-β pathway. Taken together, we propose that fascin regulates the proliferation and invasiveness of ESCC cells by modulating the expression of CTGF and CYR61 via TGF-β pathway.Esophageal cancer has the poorest prognosis among the malignant tumors of the digestive tract. Despite advancements in multimodal therapy, the overall 5-year survival rate for patients with esophageal squamous cell carcinoma (ESCC) remains poor.^1,2 In an effort to improve prognosis of ESCC, several molecular markers, such as interleukin 6 and matrix metalloproteinase 12, have been identified.^3,4 However, the molecular mechanisms underlying esophageal carcinogenesis still remain largely unknown. Fascin, an actin-bundling protein, has recently been implicated in a variety of tumors including ESCC, which promoted our investigation.Fascin is normally expressed in the mesenchymal tissues and nervous system.⁵ It is reported to function by forming parallel actin bundles in either lamellipodial or filopodial cell protrusions, which are key cellular structures for environmental guidance and cell migration.^6,7 Aberrant fascin expression has been reported in various human carcinomas such as colon cancer and gastric cancer.^8,9 Fascin is overexpressed in ESCCs, and its increased expression is associated with a poor prognosis.¹⁰ Our previous study reported the overexpression of fascin in the early stage of ESCC progression, which facilitated the proliferation and invasiveness of cancer cells.^11,12 However, the molecular basis of fascin function in the progression of ESCC remains largely unknown.In this study, first, we retrospectively examined the expression of fascin in large amounts of ESCC tissue samples by immunohistochemistry, and showed that overexpression of fascin was related to the poor survival of ESCC patients. RNAi-mediated knockdown of fascin in ESCC cells significantly inhibited cell proliferation and invasiveness, whereas forced expression of fascin in immortalized esophageal epithelial cells accelerated cell proliferation and invasiveness. Then, to explore the mechanism underlying the effects of fascin in ESCCs, we used cDNA microarrays to analyze gene expression profiles in PSC cells (ESCC cells that express high levels of fascin) and PSF8 cells (fascin-depleted ESCC cells).¹¹ Several differentially expressed genes have been identified and categorized based on their biological function. Specially, the proliferation- and invasiveness-related genes were predominantly represented. The data of cDNA array has been validated by real-time RT-PCR, Western blotting, and immunofluorescence analyses. As a result, we focused on two down-regulated genes: CYR61 (Cysteine-rich, angiogenic inducer 61) and CTGF (Connective tissue growth factor), both of which were CCN (CYR61/CTGF/NOV) members.¹³ Further studies demonstrated that CYR61 and CTGF were directly involved in fascin-mediated proliferation and invasiveness of ESCC cells. Additionally, we also revealed that fascin might affect the expressions of CYR61 and CTGF via TGF-β signaling pathway.