Expression In Lung Cancer Tissues Research Articles

Is leptin a predictive factor in patients with lung cancer?

ObjectivesThe aim of this study was to evaluate the expression and clinical significance of leptin in lung cancer. Methods126 patients with lung cancer ranged from 30 to 83years of age were studied. Serum leptin levels were determined by ELISA. The mRNA and protein levels of leptin in normal and lung cancer tissues were measured by RT-PCR and immunohistochemistry. The relationships between leptin levels and clinicopathological factors were evaluated by Wilcoxon rank sum or Kruskal–Wallis H test. ResultsSerum leptin levels in lung cancer patients were significantly higher compared to those in controls and leptin expression in lung cancer tissue was markedly increased than that in normal lung tissue (both P<0.050). ConclusionsDetermination of leptin levels might provide useful predictive information for lung cancer.

Effects of MICA Expression on the Prognosis of Advanced Non-Small Cell Lung Cancer and the Efficacy of CIK Therapy

ObjectiveTo investigate the clinical significance of the expression of MHC class I chain-related gene A (MICA) in patients with advanced non-small cell lung cancer and explore the relationship between MICA expression and the efficacy of cytokine-induced killer cell (CIK) therapy for treating advanced non-small cell lung cancer.MethodsWe obtained data on 222 patients with advanced non-small cell lung cancer, including data on MICA expression, age, gender, ECOG score, pathological type, stage, treatment history (including 38 patients who were given autologous CIK cell infusion), and overall survival (OS). MICA expression in lung cancer tissue was evaluated by immunohistochemical staining. Analyses of MICA expression, and CIK therapy association with survival outcomes were performed using Cox proportional models, Kaplan-Meier methods, and the log-rank test.Result s MICA was expressed in both membrane and cytoplasm. MICA expression correlated with the stage of lung cancer, ECOG score, gender and age. Multivariate COX regression analysis showed that the expression of MICA was an independent prognostic factor of advanced non-small cell lung cancer (p = 0.002). In subgroup analysis, we divided the 222 patients into CIK and control groups. In the CIK group, the medium OS (mOS) of patients with a high expression of MICA was longer than in those with low expression of MICA (27 months vs. 13 months). In the control group, the mOS in patients with a high expression of MICA was shorter than in patients with low MICA expression (9 months vs. 18 months). COX regression analysis showed that the MICA expression affects the effect of CIK therapy (p<0.0001).Conclusion1) The high expression of MICA is one of the indicators of a poor prognosis for advanced non-small cell lung cancer patients. 2) The high expression of MICA might be one of the predictive factors for successful CIK therapy.

Abnormal expression of Pygopus 2 correlates with a malignant phenotype in human lung cancer

BackgroundPygopus 2 (Pygo2) is a Pygo family member and an important component of the Wnt signaling transcriptional complex. Despite this data, no clinical studies investigating Pygo2 expression in lung cancer have yet been reported.MethodsIn the present study, the expression patterns of Pygo2 were evaluated by immunochemistry in 168 patients with non-small cell lung cancer (NSCLC). We used small interfering RNA (siRNA) to specifically silence Pygo2, and investigated its effect on cell growth by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis in human lung cancer cell lines.ResultsImmunohistochemical analysis showed low expression of Pygo2 in normal lung tissues and increased nuclear expression in lung cancer tissues, either with or without perinuclear expression. Abnormal Pygo2 expression was associated with poor differentiation and a high Tumor (T), Node (N) and Metastases (M) stage in NSCLC patients, and correlated with poor prognosis. Using MTT assay we observed that Pygo2 downregulation inhibited cell proliferation; in addition, flow cytometry analysis showed that Pygo2 knockdown induced apoptosis and increased numbers of G1-phase cells and a reduction in S-phase cells.ConclusionsWe therefore conclude that abnormal Pygo2 protein expression may be a marker for advanced NSCLC. Furthermore, Pygo2 knockdown suppresses cell growth.

Abstract 3070: Suppression of tumor cell invasiveness and in vivo tumor growth by microRNA-874 in non-small cell lung cancer.

Abstract MicroRNAs are a family of small non-coding RNAs that regulate expression of many genes involved in normal development as well as human disorders including cancer. Here, we show that miR-874 plays a tumor suppressor role in non-small cell lung cancer (NSCLC) in vitro and in vivo. Interestingly, in silico target prediction analysis revealed numerous pro-oncogenes associated with tumor initiation, angiogenesis, invasion and migration, including MMP-2, uPA, NFAT5, PAK3, PAK7, LIMD, PECAM1, VEGF-A, Stat3, CUX1 and FGF1 as the putative target genes of miR-874. Our preliminary in situ hybridization experiments demonstrated the diminution of miR-874 expression in lung cancer tissues as compared to its normal counterparts. Overexpression of miR-874 in CD133-positive NSCLC cancer stem cell (CSC) population led to a significant loss in CSC phenotype and enhanced sphere de-differentiation into epithelial-like morphology. Restoration of miR-874 expression drastically reduced the invasive ability of cells in comparison to mock and control-miR treated cells by suppressing the protein levels of MMP-2 and uPA. Further, miR-874 treatment decreased orthotopic tumor growth in nude mice as compared to mock and control-miR treatments. Further, in vivo orthotopic tumor experiments in nude mice revealed the significant decrease in the immunoreactivity of human anti-MMP-2 and anti-uPA in miR-874-treated tumor sections. In conclusion, our study highlights the possible tumor suppressor role of miR-874 in NSCLC-initiating cells and suggests miR-874 as a potential target in the future treatment of NSCLC. Citation Format: Divya Kesanakurti, Dilip Maddirela, Subramanyam Chittivelu, Jasti S. Rao, Chandramu Chetty. Suppression of tumor cell invasiveness and in vivo tumor growth by microRNA-874 in non-small cell lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3070. doi:10.1158/1538-7445.AM2013-3070 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

Suppression of tumor cell invasiveness and in vivo tumor growth by microRNA-874 in non-small cell lung cancer

MicroRNAs are a novel family of small non-coding RNAs that regulate the expression of several genes involved in normal development as well as human disorders including cancer. Here we show that miR-874 plays a tumor suppressor role in non-small cell lung cancer (NSCLC) in vitro and in vivo. In silico target prediction analysis revealed numerous genes associated with tumor progression including MMP-2 and uPA as the putative target genes of miR-874. Our preliminary in situ hybridization experiments demonstrated the diminution of miR-874 expression in lung cancer tissues compared to their normal counter parts. Overexpression of miR-874 in CD133-positive cancer stem cell (CSC) population led to a significant loss in CSC-phenotype and enhanced sphere de-differentiation into epithelial-like cells. Restoration of miR-874 expression drastically reduced cell invading ability in comparison to mock and control-miR-treated cells by suppressing the protein levels of MMP-2 and uPA. In in vivo experiments, miR-874 treatment decreased orthotopic tumor growth in nude mice compared to mock and control-miR treatments. Further, the immunoreactivity of human anti-MMP-2 and anti-uPA was significantly reduced in tumor sections from mice that received miR-874 treatment. In conclusion, our study highlights the possible tumor suppressor role of miR-874 in NSCLC-initiating cells and suggests miR-874 as a potential target in the treatment of NSCLC.

Proteomics Characterization of Haptoglobin Alpha-2 Subunit in Non-Small Cell Lung Cancer Serum and Various Human Materials

Non-small cell lung cancer (NSCLC) is detected in all lung cancer patients and the serum samples as potential source are often used as a potential source for early diagnosis. In this study, we used the proteomic approaches combining 2-DE, 2-D DIGE and mass spectrometry to identify the potential serum biomarkers in NSCLC serum samples, and compare the result to normal serum samples. The haptoglobin alpha 2 subunit (HAP2) is one of the proteins we are interested in and showed the overexpression level in NSCLC serum amongst the NSCLC subtypes. The identity of HAP2 was confirmed by LC-MS/MS and NH2-terminal amino acid sequencing analyses, while the high expression level of HAP2 in NSCLC serum samples was validated using co-immunoprecipitation and western blotting. Interestingly, the high expression of HAP2 has specificity in human serum samples, but nonspecific expression in lung cancer tissues and is not observed in lung cancer cell lines except HepG2 hepatocellular carcinoma cell line. This indicates that HAP2 is not produced from lung tissues and/or cells, but may be secreted from the liver. In addition, HAP2 isoforms and its post-translational modifications (PTMs) could be detected by various staining methods such as phosphoprotein, glycoprotein stains and FITC-labeled lectins stain. The different carbohydrate specificities of HAP2 isoforms in NSCLC serum samples, especially sialyl Lewis x, was observed and may correlate to the development or metastasis of lung cancer. We suggest that HAP2 may become a potentially useful serum biomarker for early diagnostic and therapeutic applications.

Influence of Quercetin-Rich Food Intake on microRNA Expression in Lung Cancer Tissues

Epidemiologic studies have reported that frequent consumption of quercetin-rich foods is inversely associated with lung cancer incidence. A quercetin-rich diet might modulate microRNA (miR) expression; however, this mechanism has not been fully examined. miR expression data were measured by a custom-made array in formalin-fixed paraffin-embedded tissue samples from 264 lung cancer cases (144 adenocarcinomas and 120 squamous cell carcinomas). Intake of quercetin-rich foods was derived from a food-frequency questionnaire. In individual-miR-based analyses, we compared the expression of miRs (n = 198) between lung cancer cases consuming high versus low quercetin-rich food intake using multivariate ANOVA tests. In family-miR-based analyses, we used Functional Class Scoring (FCS) to assess differential effect on biologically functional miR families. We accounted for multiple testing using 10,000 global permutations (significance at P(global) < 0.10). All multivariate analyses were conducted separately by histology and by smoking status (former and current smokers). Family-based analyses showed that a quercetin-rich diet differentiated miR expression profiles of the tumor suppressor let-7 family among adenocarcinomas (P(FCS) < 0.001). Other significantly differentiated miR families included carcinogenesis-related miR-146, miR-26, and miR-17 (P (FCS) < 0.05). In individual-based analyses, we found that among former and current smokers with adenocarcinoma, 33 miRs were observed to be differentiated between highest and lowest quercetin-rich food consumers (23 expected by chance; P(global) = 0.047). We observed differential expression of key biologically functional miRs between high versus low consumers of quercetin-rich foods in adenocarcinoma cases. Our findings provide preliminary evidence on the mechanism underlying quercetin-related lung carcinogenesis.

Aldehyde Dehydrogenase 1A1 Possesses Stem-Like Properties and Predicts Lung Cancer Patient Outcome

Lung cancer contains a small population of cancer stem cells that contribute to its initiation and progression. We investigated the biological function and clinical significance of aldehyde dehydrogenase 1A1 (ALDH1A1) in non-small-cell lung carcinoma (NSCLC). ALDH1A1 assay or small interfering RNA transfection was employed to isolate ALDH1A1+ cells or knock down ALDH1A1 expression in H2087 cells, respectively. Biological functions of ALDH1A1+ and ALDH1A1 silenced cells were investigated using in vitro and in vivo methods. ALDH1A1 expression was analyzed using immunohistochemistry on tissue microarrays with 179 lung cancer tissues and 26 normal lung tissues. The abilities of clone formation, proliferation, cell growth, and migration were increased in ALDH1A1+ and ALDH1A1 silenced cells. ALDH1A1+ lung cancer cells initiated tumors that resembled the histopathologic characteristics and heterogeneity of the parental lung cancer cells in mice. The silencing of ALDH1A1 expression in H2087 lung cancer cells inhibited cell proliferation and migration significantly. ALDH1A1 was expressed in 42% of normal lung tissues (11 of 26), with strong expression in the basal cells and globular cells of the normal bronchus and weak expression in the alveolar epithelial cells. Compared with normal lung tissues, 45% of NSCLC samples (81 of 179) were read as positive for ALDH1A1. Positive ALDH1A1 expression was correlated with patients' smoking status (p = 0.022), lymph-node metastasis (p = 0.006), clinical stage (p = 0.004), and a decreased overall survival time (p < 0.001). Positive ALDH1A1 expression in lung cancer tissues was an independent prognostic factor for NSCLC (odds ratio = 5.232, p < 0.001). Elucidating the biological functions of ALDH1A1 could be helpful in studying lung tumorigenesis and for developing new therapeutic approaches.

Correlation of the expression of III β-tubulin and MDR1 protein with biological features of non-small cell lung cancer

To explore the expression of III β-tubulin and MDR1 protein in patients with non-small cell lung cancer (NSCLC), and to clarify its clinical significance. Paraffin embedded tissues from 158 primary non-small cell lung cancers and para-cancerous lung tissues were investigated for the expression of III β-tubulin and MDR1 protein by immunohistochemistry, as well as in freshly-taken NSCLC tissues by Western blot. The relationship between the expression of III β-tubulin and MDR1 and the biological features of lung cancer was analyzed. The positive rate of III β-tubulin and MDR1 protein expression in lung cancer tissues was 65.19% and 51.27%, respectively. Western blot analysis showed that the level of of III β-tubulin and MDR1 protein in NSCLC tissues was remarkably higher than that in normal tissues (P < 0.01). The expression of III β-tubulin in stage III-IV cases was significantly higher than that in stage I-II cases (P < 0.05), while the expression of MDR1 protein showed no significant difference (P > 0.05). The positive rate of III β-tubulin expression in well-moderate pathological grades was lower than that in poor ones. The positive rate of MDR1 expression in adenocarcinoma was higher than that in squamous cell carcinoma and large cell undifferentiated cancers (P < 0.01). The positive rate of expression of MDR1 protein and III β-tubulin was not correlated with sex, age, tumor size and lymph node metastasis (P > 0.05). The expression of III β-tubulin and MDR1 may play an important role in the development and progression of human non-small cell lung cancer, and could be looked as an important index for judging the prognosis of lung cancer.

Abstract 198: Two novel insertion polymorphisms of the BRM gene are associated with loss of BRM expression and lung cancer risk

Abstract BACKGROUND: BRM, a catalytic subunit of the SWI/SNF chromatin remodeling complex, regulates expression/function of key signal transduction pathways with anticancer functions. In 15-25% of lung cancers, BRM protein expression is lacking. When BRM null mice (who display distinct cell cycle abnormalities) are exposed to carcinogens, they develop 10-fold more tumors then control mice. BRM silencing does not appear to be driven by mutational changes. We hypothesized that BRM silencing may be related to sequence variants in the BRM promoter region. METHODS: We compared novel BRM promoter polymorphisms to BRM protein expression in cancer cell lines and tumors of lung cancer patients. We then performed a case-control analysis of these polymorphisms with lung cancer risk. RESULTS: Sequencing of human cancer cell lines identified a 7-bp (rs34480940; located −741 from transcription start site) and 6-bp (rs3832613; located −1321) BRM insertion polymorphisms. In Caucasians, minor allele frequencies (MAF) of 45% were found for both polymorphisms. Cancer cell lines were evaluated for BRM protein expression (Western blot). Twelve BRM staining (BRM-positive) and twelve non-staining (BRM-negative) cancer cell lines were genotyped. In the BRM-negative cell lines, 11/12 cell lines were homozygous variant for at least one of the two BRM promoter polymorphisms (5/12 were double homozygous; 6/12 were homozygous for one polymorphism). In contrast, the BRM-positive cell lines yielded a good mix of genotypes for both polymorphisms. In lung cancer tissues, all ten BRM-negative samples carried at least one homozygous variant (eight were double homozygous variant, while two were homozygous for one polymorphism). In contrast, genotyping of normal adjacent tissue of twelve BRM-positive samples yielded population-normal MAFs of 42% for BRM −741 and 46% for BRM −1321. Tumour-normal tissue genotyping concordance rate was 86%. In the case-control analysis, 484 ever-smoker lung cancer cases were compared to 715 age and gender frequency-matched ever-smoker controls. Compared with a wildtype reference, carrying one homozygous variant was associated with an adjusted odds ratio (aOR) of 1.40 (95%CI:0.95-2.05; p=0.09); carrying two homozygous variants was associated with aOR=2.19 (1.40-3.43; p=0.0006), after adjusting for age, gender, and smoking variables. CONCLUSIONS: Homozygous variants of two novel BRM promoter polymorphisms are tightly associated with loss of BRM expression in lung cancer tissues and cancer cell lines. These homozygous variants are also associated with lung cancer risk in ever-smokers. Since epigenetic silencing of BRM can be reversed by various compounds (e.g. HDAC inhibitors, novel compounds from screening libraries), reversing BRM silencing could be developed as part of a chemoprevention strategy in smokers who carry the homozygous variants of these two BRM promoter polymorphisms. 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 198.

143 The prediction of lymph node metastasis in ductal carcinoma in situ with microinvasion by assessing lymphangiogenesis

The aim of this study was to investigate the mechanism of PRL-3 in inducing angiogenesis and lymphangiogenesis to promote distant and lymph node metastasis in human lung cancer tissues and cells.We investigated the expression of PRL-3, VEGF, and VEGF-C from 94 patients with non-small cell lung cancer(NSCLC) using immunohistochemical staining. The relationship between PRL-3 expression and microvessel density (MVD), lymphatic vessel density (LVD), clinicopathological factors, and surgical treatment outcome was also studied.Following this, we studied the effect on A549 by blocking PRL-3.PRL-3 expression in NSCLC was high, and this over-expression is correlated withadvanced clinical stage (p = 0.019), distant metastasis (p = 0.001), lymph node metastasis (p = 0.001), and poor post-operative survival. PRL-3 over-expression was associated with vascular endothelial growth factor (VEGF; p = 0.000) and VEGF-C(p = 0.008) expressions, MVD and LVD (p = 0.000 and p = 0.000). Blocking PRL-3 expression in A549 cell resulted in decreased cellular proliferative, migratory, and invasive abilities, and VEGF, VEGF-C, pERK, Rho-A, and Rho-C expression was inhibited. Following inhibition of either Rho or pERK, VEGF expression was down-regulated.PRL-3 induces microvascular and lymphatic vessel formation by facilitating VEGF and VEGF-C expression in lung cancer tissues, thus promoting distant and lymph node metastasis of lung cancer. PRL-3 up-regulates pERK and Rho expression and activity, facilitating VEGF expression, and accelerating angiogenesis anddistant metastasis. How to regulate VEGF-C expression needs to be further studied.

Receptor for advanced glycation end products (RAGE) soluble form (sRAGE): a new biomarker for lung cance

Receptor for advanced glycation end products (RAGE) may be involved in the pathogenesis of cancer progression. Pathological effects mediated via RAGE are physiologically inhibited by soluble RAGE (sRAGE). The aim of this study was to identify sRAGE and RAGE expression profile in lung cancer patients. An ELISA method was used to quantify serum sRAGE in 45 individuals. Additionally, surgical specimens of 28 lung cancer patients were also included for RAGE expression by immunohistochemistry. Serum sRAGE was significantly decreased in lung cancer patients compared with controls (vs. healthy donors, P=0.034; vs. pulmonary tuberculosis patients, P=0.010). Lower sRAGE concentration was negative correlated with lymph node involvement (N0 vs. N1-2, P=0.028). Down regulation of membranous and cytoplasmic expression for RAGE was also lower in lung cancer tissue than in nearby normal lung tissue. Correlation with serum sRAGE concentration and RAGE expression in lung cancer tissue was existed by CV values. The results indicate that serum sRAGE levels are decreased during lung cancer progression and could reflect decreased RAGE expression in tissue. Serum sRAGE may serve as an effective and convenient diagnostic biomarker for lung cancer. sRAGE, serum, RAGE, tissue, lung cancer.

The Clinical and Pathologic Features according to Expression of Acyl Protein Thioesterase-1 (APT1) in Stage I Non-small Cell Lung Cancer

Background: Acyl protein thioesterase-1 (APT1) is a cytosolic protein that may function in the depalmitoylation of numerous proteins, including the Ras family. However, the clinical role of depalmitoyl thioesterase in human cancer is not known. We evaluated the APT1 expression in lung cancer tissue and its clinicopathological findings according APT1 expression pattern. Methods: APT1 expression was examined by immunohistochemistry in the tumor tissue from 79 patients, who had undergone curative surgical removal of the primary lesion; all patients had been diagnosed with stage I non-small cell lung cancer between 1993 and 2004, at Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea. Results: The APT1 expression was seen in 50 out of 79 (63.3%) cases. The positive APT1 expression was significantly related with histologic subtype and T stage, but was not influenced by differentiation. The positive APT1 expression was not significantly related to patient age, gender, or smoking history. The median follow-up duration was 10.0 years; the 5-year survival rate was 71.0%. The positive APT1 expression group showed significantly worse overall survival and worse disease-free survival without statistical significance. Conclusion: We conclude that positive APT1 expression in stage I lung cancer after surgery is closely associated with overall survival. To evaluate APT1 as a prognostic marker in lung cancer, comprehensive studies on advanced stage cases are needed.

Diagnostic relevance of overexpressed Nanog gene in early lung cancers

Currently, no target molecules have been identified that enable the diagnosis of lung cancer with high sensitivity and specificity, especially in the early clinical stages of cancer. Recently, Nanog has been reported to play an important role in the self-renewal and regeneration of ES cells by maintaining these cells in the undifferentiated state and by accelerating cell proliferation. Here, we compared the degree of Nanog mRNA expression in lung cancer tissues with that in non-cancerous tissues. Nanog mRNA was detected in 84.8% (39/46) of lung cancer tissues. The sensitivity and specificity of this diagnostic technique was 80.4 and 93.3%, respectively, as estimated using the cut-off obtained from the analysis of the receiver operating characteristic curve. Further, comparison of paired cancerous and non-cancerous tissues from the same patient revealed elevated Nanog mRNA levels in all patients. No obvious correlations were detected between the clinicopathological factors and Nanog mRNA expression; however, Nanog mRNA was expressed at high levels even in the early clinical stages of the cancer. In addition, the transduction of Nanog siRNA in lung carcinoma cells resulted in growth inhibition. These results suggest that Nanog mRNA might be a new tool to support the diagnosis of lung cancers, irrespective of the clinical stage.

Promotion of MAG-1 on Metastasis of Lung Cancer Cells in vitro and Its Expression in Lung Cancer Tissue of 24 Cases.

Tumor metastasis is a multistep process with many genes involved in. A novel gene MAG-1, identified by suppression subtractive hybridization from lung cancer cells was found to be associated with tumor metastasis. The aims of this work are to investigate the metastasis related effects of MAG-1 on human lunggiant-cell line PLA801, and to compare the expression rate of MAG-1 in cancer tissue from lung cancer patients with different metastatic status. Sense and anti-sense expressing vectors of MAG-1 were constructed and transfected into PLA801C and PLA801D respectively. Colony forming, adherence assay, MTT assay and Transwell experiments were used to evaluate the alterations of clone forming, cell-matrix adherence, proliferating and invasion of the stable transfected cell strains. Western blot was employed to detect the proteins levels of CD44, and MMP-2 in cell strains and mRNA state of MAG-1 in lung cancer tissue from patients with or without pathological metastasis were also analyzed by RTPCR. MAG-1 could increase cell-ECM (Extracellular Matrix) adhesive capacity, promote invasion, enhance cell proliferation and had no effects on clone forming ability of PLA801 cells in soft agar. MAG-1 was also found have positive effects on the protein level of CD44 and MMP-2 in PLA801 cells, and the detection rate of MAG-1 mRNA was much higher in cancer tissue from metastatic patients (7/12) than that in non-metastatic patients (2/12). MAG-1 could promote lung cancer metastasis and might be a metastasis associated gene of lung cancer.

PRL-3 facilitates angiogenesis and metastasis by increasing ERK phosphorylation and up-regulating the levels and activities of Rho-A/C in lung cancer

The aim of this study was to investigate the mechanism of PRL-3 in inducing angiogenesis and lymphangiogenesis to promote distant and lymph node metastasis in human lung cancer tissues and cells. We investigated the expression of PRL-3, VEGF, and VEGF-C from 94 patients with non-small cell lung cancer (NSCLC) using immunohistochemical staining. The relationship between PRL-3 expression and microvessel density (MVD), lymphatic vessel density (LVD), clinicopathological factors, and surgical treatment outcome was also studied. Following this, we studied the effect on A549 by blocking PRL-3. PRL-3 expression in NSCLC was high, and this over-expression is correlated with advanced clinical stage (p = 0.019), distant metastasis (p = 0.001), lymph node metastasis (p = 0.001), and poor post-operative survival. PRL-3 over-expression was associated with vascular endothelial growth factor (VEGF; p = 0.000) and VEGF-C (p = 0.008) expressions, MVD and LVD (p = 0.000 and p = 0.000). Blocking PRL-3 expression in A549 cell resulted in decreased cellular proliferative, migratory, and invasive abilities, and VEGF, VEGF-C, pERK, Rho-A, and Rho-C expression was inhibited. Following inhibition of either Rho or pERK, VEGF expression was down-regulated. PRL-3 induces microvascular and lymphatic vessel formation by facilitating VEGF and VEGF-C expression in lung cancer tissues, thus promoting distant and lymph node metastasis of lung cancer. PRL-3 up-regulates pERK and Rho expression and activity, facilitating VEGF expression, and accelerating angiogenesis and distant metastasis. How to regulate VEGF-C expression needs to be further studied.

Up-regulation of the Receptor for Advanced Glycation End Product (RAGE) in Esophageal Cancer and Down-regulation in Lung Cancer and Their Relationship to Clinicopathological Features

Objective: Esophageal and lung cancers are common thoracic malignant tumors. Although the receptor for advanced glycation end product (RAGE), a newly found tumor invasion gene, has been implicated in several human cancers, the relationship between expression of RAGE and clinical features of esophageal cancer remains unknown. Moreover, no data is available for comparison between RAGE expression in common thoracic tumors and their normal tissues. The aim of the present study is to investigate RAGE expression in esophageal and lung cancers and their relationship to clinicopathological features. Methods: We quantified RAGE mRNA and protein expression levels from 12 fresh esophageal cancer and 12 lung cancer tissues by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blotting, respectively. In addition, the relationship of RAGE expression and clinicopathological features was further determined by immunohistochemistry in 29 esophageal and 32 lung cancer patients. Results: RAGE mRNA expression was increased in esophageal cancer and decreased in lung cancer compared with normal tissues. Consistent with the expression in mRNA, RAGE protein expression in esophageal cancer and lung cancer also showed the difference in tumors and normal tissues. By immunohistochemistry, cells showed membranous and cytoplastic expression for RAGE. Higher RAGE expression was more frequently detected in infiltrating than noninfiltrating esophageal tumors. Lower RAGE expression in lung cancer tissues was associated with age and UICC stages, but not with tumor differentiation, histological typing, and lymph node metastasis. Conclusions: Expression of RAGE and relationships with their clinicopathological features were different in the thoracic tumors. RAGE was increased in esophageal cancer, especially in infiltrating tumors, but decreased and was closely associated with age and UICC stages in lung cancers. These results suggest that RAGE might be involved in the development of esophageal and lung cancers.

Prevention of chinese green tea on 3,4-benzopyrene-induced lung cancer and its mechanism in animal mode.

Chinese green tea is one of the daily consumption beverages in the world and is considered a promising cancer chemopreventive agent. In the present study, we investigate the role of lung cancer prevention by green tea and its mechanism. Three groups of female SD rats were kept with the same feed. Rats in group A were administrated with 1% green tea drinking, while in group B and group C with water only. Animals in group A and group B were given 3,4-benzopyrene-corn oil mixture pulmonary injection fortnightly for 4 times, while in group C corn oil only. Rats were sacrificed 1 year after the first injection under narcotism. Lung tumors and lung tissues were performed H&E staining for cancer identification. Each case of lung cancer was examined for expression of p53 and Bcl-2 with in situ hybridization analysis and immunohistochemistry staining. No cancer was found in rats in group C. However, in group B, 15 out of 20 rats were found generating lung cancer, and in group A, 6 out of 20 rats inducing lung cancer were recorded. The rate of lung carcinogenesis in rats was decreased from 75% to 30% by 1% chinese green tea oral administration (Chi-Square=8.12, P <0.01). Higher level of p53 expression in lung cancer tissues of group A was observed under microscope than that of group B, but the difference has no statistic significance (P >0.05). However, significantly lower level of Bcl-2 expression was found in lung cancer tissues of group A than that of group B (P <0.05). The results indicate that chinese green tea inhibits lung carcinogenesis. Chinese green tea can slightly upregulate expression of p53, but significantly downregulate expression of Bcl-2 in lung cancer, and this may be related to the mechanism of lung cancer prevention.

HnRNP K expression and its clinical significance in human lung cancer tissues.

There is no any specific criterion in early diagnosis of lung cancer yet. The aim of this study is to determine hnRNP K expression in lung cancer tissues and normal lung tissues and to provide data for the diagnosis and treatment of lung cancer. The expression of hnRNP K was studied by using the SP method of immunohistochemistry in lung tissue samples of 70 cases of lung cancer and 27 cases of control groups. The positive rate of hnRNP K was 64.9% (76/117) in lung cancer tissues , which was significantly higher than that in control group (33.3%, 9/27) (P <0.001) . hnRNP K was localized in the cytoplasm and/or nucleus. Three subtypes of lung cancer all showed positive staining of hnRNP K protein. The positive rate of hnRNP Kwas 73.7% (14/19) in SCLC and 54.9% (28/51) in NSCLC (P >0.05) . hnRNP K can highly express in lung cancer tissues, but it does not correlate with histological classification of lung cancer.

VILIP-1 Downregulation in Non-Small Cell Lung Carcinomas: Mechanisms and Prediction of Survival

VILIP-1, a member of the neuronal Ca++ sensor protein family, acts as a tumor suppressor gene in an experimental animal model by inhibiting cell proliferation, adhesion and invasiveness of squamous cell carcinoma cells. Western Blot analysis of human tumor cells showed that VILIP-1 expression was undetectable in several types of human tumor cells, including 11 out of 12 non-small cell lung carcinoma (NSCLC) cell lines. The down-regulation of VILIP-1 was due to loss of VILIP-1 mRNA transcripts. Rearrangements, large gene deletions or mutations were not found. Hypermethylation of the VILIP-1 promoter played an important role in gene silencing. In most VILIP-1-silent cells the VILIP-1 promoter was methylated. In vitro methylation of the VILIP-1 promoter reduced its activity in a promoter-reporter assay. Transcriptional activity of endogenous VILIP-1 promoter was recovered by treatment with 5′-aza-2′-deoxycytidine (5′-Aza-dC). Trichostatin A (TSA), a histone deacetylase inhibitor, potently induced VILIP-1 expression, indicating that histone deacetylation is an additional mechanism of VILIP-1 silencing. TSA increased histone H3 and H4 acetylation in the region of the VILIP-1 promoter. Furthermore, statistical analysis of expression and promoter methylation (n = 150 primary NSCLC samples) showed a significant relationship between promoter methylation and protein expression downregulation as well as between survival and decreased or absent VILIP-1 expression in lung cancer tissues (p<0.0001). VILIP-1 expression is silenced by promoter hypermethylation and histone deacetylation in aggressive NSCLC cell lines and primary tumors and its clinical evaluation could have a role as a predictor of short-term survival in lung cancer patients.