Expression In Lung Carcinoma Research Articles

Therapeutic efficacy of ECs Foxp1 targeting Hif1α-Hk2 glycolysis signal to restrict angiogenesis

Endothelial cells (ECs) rely on glycolysis for energy production to maintain vascular homeostasis and the normalization of hyperglycolysis in tumor vessels has recently gained attention as a therapeutic target. We analyzed the TCGA database and found reduced Foxp1 expression in lung carcinoma. Immunostaining demonstrated reduced expression more restricted at tumor vascular ECs. Therefore, we investigated the function and mechanisms of Foxp1 in EC metabolism for tumor angiogenesis required for tumor growth. EC-Foxp1 deletion mice exhibited a significant increase of tumor and retinal developmental angiogenesis and Hif1α was identified as Foxp1 target gene, and Hk2 as Hif1α target gene. The Foxp1-Hif1α-Hk2 pathway in ECs is important in the regulation of glycolytic metabolism to govern tumor angiogenesis. Finally, we used genetic deletion of EC-Hif1α and RGD-peptide nanoparticles EC target delivery of Hif1α/Hk2-siRNAs to knockdown gene expression which reduced the tumor EC hyperglycolysis state and restricted angiogenesis for tumor growth. This study advances our understanding of EC metabolism for tumor angiogenesis, and meanwhile provides evidence for future therapeutic intervention of hyperglycolysis in tumor ECs for suppression of tumor growth.

P8‐68: Development of a potential biomarker for PD‐L1 expression in lung carcinoma

P8‐68: Development of a potential biomarker for PD‐L1 expression in lung carcinoma

PDL-1 expression in lung carcinoma and its correlation with clinicopathological and prognostic characteristics

ABSTRACT Lung cancers have high incidence and high mortality rates. The immune checkpoints as programmed death ligand 1 (PDL-1) can suppress the tumor immune reaction. So, their blocking seems to be a way to treat tumors. This study assesses PDL-1 immunohistochemical expression in lung cancer, and its correlation with prognosis. It included 62 specimens of lung cancer in Hospitals of Mansoura Faculty of Medicine, Egypt. Seventy-one percent of cases showed positive PDL-1 and about 59.1% of them showed high expression. PDL-1 expression in NSCLC was significantly higher than in SCLC (P = 0.019). There were no significant associations between PDL-1 expression and other clinicopathological parameters. A significant mild positive correlation between PDL-1 and EGFR marker was found (P = 0.006). The mean overall survival in cases with positive PDL-1 was lower than negative cases (P = 0.37). Also, progression-free survival was lower among PDL-1 positive cases compared to negative cases (P = 0.5). This study reports that immune checkpoint, PDL-1 is overexpressed in lung cancer especially NSCLC. It is correlated with EGFR overexpression. PDL-1 could have potential to be an effective immune target for lung cancer immunotherapy. But the presence of PD-L1-negative tumors highlights the importance of searching for alternative or combination treatment strategies. Abbreviations: AC: Adenocarcinoma; COPD: Chronic Obstructive Pulmonary Diseases; CTLA-4: Cytotoxic T-lymphocyte-associated protein 4; EGFR: Epidermal Growth Factor Receptor; IHC: Immunohistochemical; NSCLC: Non-Small Cell Lung Cancer; OS: Overall Survival; PD1: Programmed Death 1; PDL-1: Programmed Death ligand 1; PFS: Progression Free Survival; SCC: Squamous cell carcinoma; SCLC: Small Cell Lung Cancer; SD: Standard Deviation; TCR: T-cell receptor; TPS: Tumor Proportion Score.

Applicability of pan-TRK immunohistochemistry for identification of NTRK fusions in lung carcinoma

In the last two decades, various therapies have been introduced for lung carcinoma patients, including tyrosine-kinase inhibitors for different mutations. While some of them are specific to specific tumor types, others, like NTRK1–3 fusions, are found in various solid tumors. The occurrence of an NTRK1,2 or 3 fusion acts as a biomarker for efficient treatment with NTRK inhibitors, irrespectively of the tumor type. However, the occurrence of the NTRK1–3 fusions in lung carcinomas is extremely rare. We performed a retrospective analysis to evaluate the applicability of immunohistochemistry with the pan-TRK antibody in the detection of NTRK fusions in lung carcinomas. The study cohort included 176 adenocarcinomas (AC), 161 squamous cell carcinomas (SCC), 31 large-cell neuroendocrine carcinomas (LCNEC), and 19 small cell lung carcinomas (SCLC). Immunohistochemistry (IHC) was performed using the pan-TRK antibody (clone EPR17341, Ventana) on tissue microarrays, while confirmation for all positive cases was done using RNA-based Archer FusionPlex MUG Lung Panel. On IHC staining, 12/387 samples (3.1%) demonstrated a positive reaction. Ten SCC cases (10/161, 6.2%), and two LCNEC cases (2/31, 6.5%) were positive. Positive cases demonstrated heterogeneous staining of tumor cells, mostly membranous with some cytoplasmic and in one case nuclear pattern. RNA-based sequencing did not demonstrate any NTRK1–3 fusion in our patients’ collective. Our study demonstrates that pan-TRK expression in lung carcinoma is very low across different histologic types. NTRK1–3 fusions using an RNA-based sequencing approached could not be detected. This stresses the importance of confirmation of immunohistochemistry results by molecular methods.

Non-small cell lung carcinomas with a minor sarcomatoid component and pleomorphic carcinomas are associated with high expression of programmed death ligand 1

Pleomorphic carcinomas are known to be highly programmed death ligand 1 (PD-L1) positive non-small cell lung cancer (NSCLC) types. However, the level of PD-L1 expression in lung carcinomas with a minor sarcomatoid component, comprising less than 10 % of the tumor mass, has not been determined yet. We hypothesized that NSCLC with a minor sarcomatoid component is more closely related to pleomorphic carcinomas in terms of PD-L1 expression than to NSCLC types without sarcomatoid features. The surgical resections from 690 lung carcinoma patients were retrospectively analyzed for the presence of PD-L1 by means of immunohistochemistry using the 22C3 PharmDx assay. The tumor proportion score system was applied to quantify the level of PD-L1 expression. Membranous staining present in ≥ 1 % of tumor cells was chosen as the cut-off to define a positive result for PD-L1 expression. Tumors were allocated into one of four subgroups: “adenocarcinoma”, “squamous cell carcinoma”, “pleomorphic carcinoma”, or “NSCLC with a minor sarcomatoid component”. PD-L1 expression in pleomorphic carcinomas (26/32, 81.3 %) and in the subgroup of NSCLC with a minor sarcomatoid component (35/46, 76.1 %) was identified in a comparable proportion of cases. Pleomorphic carcinomas were significantly more often PD-L1 positive than adenocarcinomas (p < 0.001) or squamous cell carcinomas (p = 0.0015). Accordingly, the proportion of PD-L1 expressing NSCLC with a minor sarcomatoid component was significantly higher than that of the adenocarcinoma (p < 0.001) or squamous cell carcinoma (p = 0.002) subgroup. In summary, we identified a presumable new subgroup of highly PD-L1 positive neoplasms within the NSCLC spectrum that is related to pleomorphic carcinomas in terms of PD-L1 expression. Further investigation regarding genetic relation and mechanism of PD-L1 expression in these two NSCLC categories is recommended.

CD117 expression is correlated with poor survival of patients and progression of lung carcinoma:a meta-analysis with a panel of 2645 patients.

The aim of this research was to investigate the clinical role and prognostic value of CD117 expression assessed immunohistochemically in lung carcinoma through acomprehensive meta-analysis in which 27 publications were acquired and 2645patients were ultimately analysed. Statistical analysis and corresponding plots were performed using STATA version 12.0. Publication bias was assessed by Begg's funnel plots and Egger's test. Pooled HR and its 95% CI (HR=1.53, 95% CI: 1.13-2.07, p=0.007) for overall survival of patients indicated apoor prognostic value for CD117 expression in lung carcinoma, which was accompanied by heterogeneity and publication bias. In the subgroup analysis, there was strong evidence that could support an association between CD117 expression and poor prognosis in NSCLC patients (HR=2.03, 95% CI: 1.41-2.90, p<0.001; heterogeneity: I2= 41.9%, c2=15.49, p=0.078). Multivariate analysis also revealed consistent results in high-quality studies with reported HRs (HR=2.16, 95% CI: 1.67-2.79, p<0.001), and Asian patients (HR=2.12, 95% CI: 1.45-3.10, p<0.001). The correlations between CD117 expression and age, clinical stage, TNM stage, lymph node metastasis, or histology were not statistically significant. In conclusion, CD117 expression might be apotential marker for predicting poor prognosis, faster tumour growth, and early lymph node metastasis in NSCLC.

GATA-3 is superior to GCDFP-15 and mammaglobin to identify primary and metastatic breast cancer.

Despite numerous studies on the utility of GATA-3 as breast cancer marker, its comparison with other breast markers, its concordance between primary and metastatic tumors and its expression in primary cancers from sites with frequent breast metastases remains unclear. To address these questions, totally 993 invasive breast cancers (IBC), 254 paired nodal metastases, 23 distant metastases, and 208 lung carcinomas were included. GATA-3 expression was analyzed by immunohistochemistry and compared to other breast markers [gross cystic disease fluid protein 15 (GCDFP-15) and mammaglobin (MGB)]. GATA-3 was expressed in 82.5% of IBC, predominantly in luminal (93.9%), and lower in non-luminal cancers [59.6% of HER2 overexpressing (HER2-OE) and 38.1% of triple negative breast cancer (TNBC) subtypes]. GATA-3 identified more IBC than GCDFP-15 (23.9%) and MGB (46.6%). However, MGB showed a comparable sensitivity for non-luminal cancers to GATA-3. Combining MGB and GATA-3 improved sensitivity for both HER2-OE (80.8%) and TNBC cases (55.4%). GATA-3 showed a high sensitivity for nodal metastases and distant metastases, with good concordance with primary tumors. GATA-3 was expressed in 1.0% of lung carcinomas, with sensitivity and specificity of 82.5 and 99.0% in differentiating IBC and lung carcinoma. GATA-3 expression was the highest in luminal breast carcinomas, and showed higher sensitivity than GCDFP-15 and MGB. However, in the poorly differentiated IBC, its utility was still limited. One should be aware of the possible GATA-3 expression in lung carcinomas.

Correlation between histone acetylation and expression of Notch1 in human lung carcinoma and its possible role in combined small-cell lung carcinoma

Combined small-cell lung carcinoma (cSCLC) is composed of small-cell lung carcinoma (SCLC) admixed with non-small-cell lung carcinoma (NSCLC). Evaluating the molecular differences between SCLC and NSCLC could lead to a better understanding of the pathogenesis of such neoplasms. Therefore, in this study, we investigated the correlation between histone acetylation and Notch1 expression in lung carcinoma. Using chromatin immunoprecipitation (ChIP) assay, we measured the level of acetylated histone H3 around the promoter region of Notch1 in SCLC and NSCLC cells. We then treated SCLC cells with trichostatin A (TSA) and characterized the level of histone H3 acetylation at Notch1. In addition, TSA-treated cells were injected into immune-compromised mice, for analysis of the ex vivo tumor xenograft phenotype. The level of acetylated histone H3 surrounding the Notch1 promoter was lower in lung cancer cells not expressing Notch1. Tumors originated from TSA-treated SCLC cells occasionally formed an epithelial-like glandular arrangement of cells; with Notch1 expression and decreased expression of neuroendocrine (NE) markers. Histone deacetylation around the promoter region of Notch1 inhibits Notch1 protein expression in SCLC and the restoration of Notch1 expression in SCLC leads to the concurrent appearance of epithelial-like areas within the SCLC, which could provide a possible mechanism for histogenesis of cSCLC.

GASC1 expression in lung carcinoma is associated with smoking and prognosis of squamous cell carcinoma.

GASC1 (gene amplified in squamous cell carcinoma 1) encodes a nuclear protein that epigenetically catalyses the lysine demethylation of histones. We investigated the expression of GASC1 in different histological subtypes of lung cancer (n=289). Percentage value of GASC1 immunohistochemical expression was evaluated separately in the nuclei and cytoplasms of epithelial cancer cells. The results were compared with clinicopathologic factors and the smoking history of the patients. In lung tumor cells, 38% of nuclei and 54% of the cytoplasms stained positive for GASC1. Adenocarcinomas expressed more GASC1 nuclear (p=0.00011) and cytoplasmic (p=0.00074) positivity than squamous cell carcinoma. Smokers displayed less nuclear and cytoplasmic GASC1 expression than non-smokers (p=0.028 and p=0.036, respectively). Similarly, patients with more cytoplasmic positive staining had fewer pack years (p=0.043). Nuclear GASC1 expression had an impairing effect on survival when all histological lung cancer types were analysed together (p=0.039) and separately in squamous cell lung carcinoma (p=0.016). The results reveal that GASC1 expression is higher in adenocarcinoma than squamous cell carcinoma. Smoking decreases GASC1 expression in tumor cells, indicating that tobacco smoke may influence the methylation of histone 3 lysine residues in lung cancer. Nonetheless, nuclear GASC1 predicts a poor prognosis, especially in squamous cell carcinoma.

Clinicopathological and prognostic significance of ERα and ERβ expression in lung carcinomas: a tissue microarray study

To detect the expression of ERα and ERβ in lung carcinomas and investigate their clinicopathological and prognostic significance by using tissue microarray assay and immunohistochemical staining. Six hundred and ninety-eight lung cancer specimens were used in this study, including 651 cases of non-small cell lung carcimomas (NSCLCs) and 47 cases of small cell lung cancers (SCLCs). There were 309 cases of adenocarcimoma and 342 cases of squamous cell carcinoma. The expression of ERα and ERβ was analyzed by immunohistochemistry on paraffin-embedded sections. In the normal lung tissues, expression of ERα and ERβ was 0% (0/35) and 25.0% (9/36), respectively. In the tumor tissues, ERα was expressed in 209 of 295 AC cases (70.8%), 169 of 330 SCC cases (51.2%) and 9 of 47 SCLC cases (19.1%) (P < 0.001). ERβ was expressed in 200 of 297 AC cases (67.3%), 140 of 322 SCC cases (43.5%) and 31 of 47 SCLC cases (66.0%) (P < 0.001). In NSCLC, the expression of ERα and ERβ was significantly associated with smoking, stage and lymph node metastasis, also with sex refer to ERβ (P < 0.05), but not significantly with age, tumor size and degree of differentiation (P > 0.05). Follow-up was completed in 398 NSCLC cases, and no significant correlation was found between the prognosis and expression of ERα and ERβ. The expression of ERα and ERβ has significant difference in lung adenocarcinoma, squamous cell carcinoma and small cell lung cancer. In NSCLC, expression of ERα and ERβ is associated with smoking, stage, and lymph node metastasis. The expression of ERβ is higher in female than in male NSCLC patients.

HIF-1alpha in lung carcinoma: Histopathological evidence of hypoxia targets in patient biopsies

Background: Hypoxia inducible factor 1alpha (HIF-1alpha) plays a major role in the response of tumors to hypoxia, and contributes to tumor aggressiveness, invasiveness and resistance to radiotherapy and chemotherapy. Targeting HIF-1alpha is an attractive strategy, with the potential for disrupting multiple pathways crucial for tumor growth. Thus, the evaluation of HIF-1alpha in patient biopsies could be useful in personalized cancer treatment. Methodology: The current literature on HIF-1alpha immunohistochemical expression is reviewed along with the relation to clinical outcome and prognosis. In addition, the significant correlation of HIF-1alpha to vascular endothelial growth factor (VEGF) expression is reported, as well as the possible role of HIF-1alpha in predicting the therapeutic response to anti-EGFR therapies. Conclusion: Herein, an overview of the HIF-1alpha expression in lung carcinoma is presented. Since there is no consensus regarding the assessment of HIF-1alpha in tissue specimens, heterogeneous results have been reported especially regarding prognosis. In this context, methods to optimize the evaluation of HIF-1alpha in biopsies are needed in order to clarify the role of HIF-1alpha as a prognostic or predictive biomarker in lung carcinoma.

Regulation of neutrophil gelatinase-associated lipocalin expression by C/EBPβ in lung carcinoma cells

Neutrophil gelatinase-associated lipocalin (NGAL), a member of the lipocalin family, has been found to be overexpressed in a variety of tumors, including lung adenocarcinomas. However, the mechanism by which NGAL expression is regulated in lung carcinoma needs further evaluation. In this study, immunohistochemistry was employed to analyze the expression of NGAL in lung carcinoma tissue samples, including lung squamous carcinomas, adenocarcinomas, adenosquamous carcinomas and bronchial alveolar cell carcinomas. The results showed that NGAL was expressed in 82.61% (19/23) of the samples. RT-PCR and immunofluorescent staining showed that NGAL was localized to the cytoplasm in lung carcinoma cell lines. To explore the transcriptional regulation mechanism of NGAL basal expression in lung carcinoma, a 1515-bp fragment (-1431 to +84) of the NGAL promoter region was cloned and a series of deletion and mutation constructs were generated. These constructs were analyzed using the luciferase reporter assay. The results indicated that the cis-acting elements important for the basal activity of NGAL transcription were likely located between -152 and -141. Further analysis using site-directed mutagenesis and the luciferase reporter assay suggested that the C/EBP binding sites were responsible for the activity of the NGAL promoter. Finally, the binding ability and specificity of the transcription factors were determined by electrophoretic mobility-shift assay (EMSA). The results showed that C/EBPβ was able to bind to the -152 and -141 segments. Taken together, these findings suggest that NGAL is expressed in lung carcinomas and that NGAL expression is mediated by the binding of C/EBPβ to the -152 and -141 segment of the NGAL promoter.

Significance of Clinicopathology and Expression of Heat Shock Protein 72 in Human Primary Lung Carcinomas

Heat shock protein 72 (HSP72) and glycoprotein 96 (gp96) are highly expressed in cancer tissues. Recent studies indicate the possible roles of HSP72 and gp96 in the development and progression of lung carcinomas but detailed information is still ambiguous. The aim of the study is to investigate the correlation between clinicopathology and immunolocalization of HSP72 and gp96 in human lung carcinoma. Immunohistochemistry demonstrated that HSP72 and gp96 expression in lung carcinomas with metastasis was significantly higher than those with non-metastasis. HSP72 and gp96 expression were significantly associated with the presence of tumor infiltration, lymph node and remote metastasis.

Significance of Clinicopathology and Expression of Heat Shock Protein 72 in Human Primary Lung Carcinomas

Heat shock protein 72 (HSP72) and glycoprotein 96 (gp96) are highly expressed in cancer tissues. Recent studies indicate the possible roles of HSP72 and gp96 in the development and progression of lung carcinomas but detailed information is still ambiguous. The aim of the study is to investigate the correlation between clinicopathology and immunolocalization of HSP72 and gp96 in human lung carcinoma. Immunohistochemistry demonstrated that HSP72 and gp96 expression in lung carcinomas with metastasis was significantly higher than those with non-metastasis. HSP72 and gp96 expression were significantly associated with the presence of tumor infiltration, lymph node and remote metastasis.

Correlation between clinicopathology and expression of heat shock protein 72 in human primary lung carcinomas

Heat shock protein 72 (HSP72) is highly expressed in cancer tissues. Recent studies indicate the possible roles of HSP72 in the development and progression of different carcinomas, but detailed information is still ambiguous. The aim of the study is to investigate the correlation between clinicopathology and immunolocalization of HSP72 in human primary lung carcinoma. The expression of HSP72 was studied in 96 human primary lung carcinomas with or without metastasis, as well as in tissues adjacent to cancer by way of immunohistochemistry. The distribution of HSP72 immunoreactive was examined by continuous cell counting in restricted fields by pathology photograph analysis. HSP72 immunoreactivities were detected in 90 of 96 primary tumors (93.7%). The expression of HSP72 has a correlation with the differentiation of primary lung carcinoma. HSP72 expression in lung carcinomas with lymph node and organ metastasis was significantly higher than those with non-metastasis. The results indicate that there exists a significant correlation between the expression of HSP72 and the progression of primary lung carcinomas. HSP72 expression were significantly associated with the presence of tumor infiltration, lymph node and remote metastasis. The expression characters of HSP72 in primary lung carcinoma may contribute to study the pathogenesis and progression of lung carcinoma. Key words: Heat shock protein 72, primary lung carcinoma, clinicopathology, prognosis.

Abstract LB-269: Genome-wide expression profiling of highly degraded formalin-fixed paraffin-embedded tissue from non-small cell lung carcinoma

Abstract Microarray-based genome-wide gene expression analysis on fresh frozen tissue is widely used in cancer studies. Compared to fresh frozen tissues, formalin-fixed paraffin-embedded (FFPE) tissues are more accessible in the clinical setting for genomic analysis, but its use has been hampered by technical difficulty. In this study, we investigated the potential use of the Whole-Genome DASL HT Assay (Illumina, San Diego, CA) for global expression profiling (covering &amp;gt;47,000 annotated transcripts) in 24 tumor/non-affected FFPE tissues obtained by biopsy from patients with non-small cell lung cancer (NSCLC). We obtained high-quality microarray data on 11 FFPE samples (46%). The FFPE samples had a relatively lower median call rate of 12.2% (range: 10–14.5%), as compared with those from fresh samples; ranging from 10% to 25%. Among genes differentially expressed between squamous carcinomas and adjacent non-affected lung tissue, we found that the RECK gene, encoding a tumor suppressor of reversion-inducing cysteine-rich protein with Kazal motifs, was decreased strongly in tumors (fold change: -2.7; p=0.018). This finding was consistent with previous reports that there was a significantly lower expression of RECK mRNA in NSCLC tissue in samples with promoter hypermethylation. During pathway-based analysis using GeneGo database, we found that differentially expressed genes were significantly enriched in several pathways, such as the RAS regulation pathways, epithelial-to-mesenchymal transition pathway, WNT signaling pathway, and signal transduction pathway of lung neoplasms. When comparing expression between adenocarcinoma and squamous NSCLC, the differentially expressed genes were clustered in keratin filaments pathway for cytoskeleton remodeling, WNT signaling pathway, and disease pathways related to bronchial neoplasms and NSCLC. The WNT/PCP signaling pathway has been implicated in human carcinogenesis. MAP3K7 is one of the key signaling molecules of non-canonical WNT pathway, which demonstrated high level expression in lung carcinoma but was not expressed in non-affected pulmonary tissues. Our study demonstrated that DASL expression profiling technology, genome-wide expression profiles can be successfully generated from FFPE lung tissues. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-269. doi:10.1158/1538-7445.AM2011-LB-269

Neutrophil interactions with sialyl Lewis X on human nonsmall cell lung carcinoma cells regulate invasive behavior

The carbohydrate sialyl Lewis X (sLeX) is expressed on leukocytes and carcinoma cells and binds to selectins during inflammatory processes and early metastasis. Synthesis of sLeX depends on activity of enzymes, including α(1,3/1,4) fucosyltransferase (FucT-III). Tumor necrosis factor-α (TNF-α) up-regulates FucT-III, resulting in increased sLeX in the airways of patients with respiratory disease; however, the mechanisms that regulate sLeX in the inflammatory tumor microenvironment are not well understood. The authors stably transfected human lung carcinoma cell lines with the FucT-III gene and exposed them to TNF-α to investigate its role in regulation of sLeX expression and selectin-binding ability using semiquantitative real-time polymerase chain reaction and flow cytometry. Cytokine expression was examined in transfected cells using chemiluminescent arrays and enzyme-linked immunosorbent assays, and invasion was studied using Matrigel assays and alterations in morphology. Human lung tissue arrays were analyzed for immunohistochemical detection of sLeX and neutrophils. Stimulation of FucT-III-transfected cells with recombinant human (rh) TNF-α up-regulated sLeX expression and increased E-selectin binding. Transfected cells secreted high levels of interleukin 8, growth-regulated oncogene-α, and mast cell proteinase-1. Cells exposed to rhTNF-α, neutrophil-conditioned media, and cultures with a 5:1 ratio of neutrophils to cancer cells had significantly increased sLeX expression and invasiveness and underwent nonadherent morphologic changes. In lung carcinomas, but not in normal lung tissues, 71% of tumors were highly positive for sLeX expression in areas of increased neutrophil infiltration. The current results indicated that neutrophils may be recruited to areas of FucT-III activity and sLeX expression in lung carcinomas to enhance the invasive and metastatic potential of lung cancer cells.

Abstract 2339: Stimulation of sialyl Lewis X carbohydrate expression in lung carcinoma cells by neutrophils: Implications for metastasis

Abstract Sialyl Lewis X (sLeX) is a carbohydrate epitope expressed on leukocytes and carcinoma cells that promotes binding of both cell types to vascular endothelium by selectin mediated processes. These binding events are crucial for leukocyte trafficking to lymphoid tissues and sites of inflammation and metastasis of cancer cells. Increased sLeX expression is correlated with advanced disease and a poor prognosis for several types of carcinomas. Synthesis of sLeX is dependent on activity of the enzymes α-(1,3)-fucosyltransferase (FucT-III) and α2,3-sialyltransferase. Exposure to tumor necrosis factor alpha (TNF-α) has been described to upregulate FucT-III resulting in increased sLeX in the airways of patients with respiratory disease, however, little is known about the molecular mechanisms involved in regulation of sLeX expression in the inflammatory tumor microenvironment. We stably transfected lung carcinoma cells with FucT-III to investigate the role of TNF-α in regulation of sLeX expression, selectin binding ability, and metastatic potential of lung carcinoma cells. Stimulation of FucT-III transfected but not mock-transfected H1299 and A549 lung carcinoma cells with recombinant TNF-α resulted in a concentration dependent upregulation of sLeX expression, increased binding of carcinoma cells to E-selectin, and increased migration of transfectants through Matrigel. Lung carcinoma cells transfected with FucT-III demonstrated high secretion of the neutrophil chemoattractant cytokines IL-8, GRO-α and MCP-1. Co-culture of lung carcinoma cells with neutrophil conditioned media and neutrophils isolated from blood at an optimal ratio of 1:5 resulted in significantly increased sLeX expression (p = 0.02 and p = 0.002 respectively). Exposure of transfected carcinoma cells to neutrophil conditioned media and neutrophils resulted in morphological changes consistent with increased invasive potential and a TNF-α dependent increased cellular migration through Matrigel. In patient lung carcinoma specimens but not in normal lung tissues, we observed areas of high sLeX expression that correlated with increased neutrophil infiltration in 80% of tumors. We conclude that FucT-III activity and sLeX expression in lung carcinomas may recruit neutrophils in the tumor microenvironment that enhance the metastatic potential of lung tumor cells. This work was supported in part by funds from the National Institutes of Health/National Cancer Institute grant K08 CA111829-04. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2339.

Comparison of Osteopontin, β-catenin and hnRNP B1 Expression in Lung Carcinomas

This study was performed to compare osteopontin (OPN), beta-catenin and heterogeneous nuclear ribonucleoprotein B1 (hnRNP B1) immunreactivities in small cell lung carcinomas (SCLC) and non-small cell lung carcinomas (NSCLC). Correlation of these three antibodies with grade and clinicopathologic stage of the tumor in NSCLC was also investigated. Twenty-nine SCLC, 6 large cell carcinoma, 36 adenocarcinoma and 30 squamous cell carcinoma (SCC), totally 101 cases, were included in this study. OPN, beta-catenin and hnRNP B1 expressions were immunohistochemically evaluated. OPN positivity was 6.9% in SCLC and 67% in NSCLC. When NSCLC types were individually considered, OPN positivity was 66.7% in large cell carcinoma, 80% in SCC and 55.6% in adenocarcinomas. beta-catenin positivity was observed in 48.6% of NSCLC and none of SCLC cases. These results were statistically significant (p < 0.05). Neither grade nor stage of NSCLC was correlated with osteopontin, beta-catenin or hnRNP B1 immunreactivity. We observed that OPN and beta-catenin are useful in differentiating SCLC from NSCLC. This may be helpful in small lung biopsies where morphology is obscured by crush artifacts.

Caveolin‐1 expression in lung carcinoma varies according to tumour histotype and is acquired de novo in brain metastases

To study caveolin-1 (Cav-1) expression in metastatic lung carcinomas. Cav-1 expression was investigated in a series of 121 lung carcinomas and it was shown that 18/121 tumours (14.9%) were Cav-1+. None of the pure bronchioloalveolar carcinomas proved to be positive, vs. 42.8% of the large cell carcinomas (neuroendocrine subtype excluded). Adenocarcinomas (8.5%), large cell neuroendocrine carcinomas (20%) and squamous cell carcinomas (29.6%) displayed an intermediate percentage of positive cases, suggesting a gradient of Cav-1 expression according to tumour histotype-related aggressiveness. Moreover, the percentage of Cav-1+ tumours with distant metastases was almost double that of non-metastatic tumours (17.8% vs. 8.1%), irrespective of the histotype. In 34 tumours metastatic to the brain, primary and secondary lesions were compared and 53% of brain metastases were Cav-1+ vs. 20.6% of primaries, indicating a de novo acquisition of Cav-1 expression. This pattern was exclusive to the brain, as it was not acquired in adrenal metastases. In our series, the presence of epidermal growth factor receptor amplification, determined by fluorescence in situ hybridization, was not related to Cav-1 reactivity. Cav-1 immunoreactivity in lung carcinoma is histotype-dependent and acquired de novo in brain metastases, suggesting a site-specific phenotypic shift in secondary lesions.