Non-cancerous Lung Tissues Research Articles

Amplification, up-regulation and over-expression of C3G (CRK SH3 domain-binding guanine nucleotide-releasing factor) in non-small cell lung cancers.

The Ras-CRK-Rap1 cellular signal-transduction system is regulated by guanine nucleotide exchange factors (GEFs). Transcription of C3G on chromosome 9q34 and a key member of the GEF gene family is activated by the CRK-adaptor protein; the C3G product is a CRK SH3 domain-binding guanine nucleotide-releasing factor. We document here the amplification of C3G in five of 18 primary non-small cell lung cancers examined and its increased expression in 18 of 28 tumors in comparison to corresponding non-cancerous lung tissues. Immunohistochemical staining revealed prominent C3G protein in the cytoplasm of cancer cells, associated with faint staining at the nucleolar membrane, but C3G was not detectable in normal bronchial mucoepithelial cells or in broncholoalveolar cells of the bronchial/bronchiolar ducts or alveoli. These data indicate that amplification and increased expression of the C3G gene may play some role in human lung carcinogenesis through derangement of the CRK-Rap1 signaling pathway.

Clinical significance of heparanase activity in primary resected non-small cell lung cancer

Clinicopathological significances of heparanase activity in non-small cell lung cancer (NSCLC) were investigated by analyzing 76 resected specimens of NSCLC. Heparanase activities in NSCLC were significantly higher than non-cancerous lung tissues ( P<0.0001). The heparanase activities of NSCLC were significantly higher in larger diameter tumors ( P=0.0141) or with metastasis to ipsilateral mediastinal lymph nodes ( P=0.0004). The activities of heparanase in primary tumors were increased significantly according to the pathological stage of the progression of the disease ( P=0.0009). Among the clinicopathological parameters, histological cell type and evidence of ipsilateral lymph node metastasis showed a significant association with elevated heparanase activities, whereas age, degree of differentiation and tumor diameter did not. Kaplan–Meier curves for overall and disease-free survival demonstrated a significant difference between patients with elevated and non-elevated heparanase activity by log–rank test ( P=0.0145 and 0.0002, respectively). Multivariate analysis showed heparanase activity was an independent factor to influence disease-free survival in our study. These results suggest that heparanase activity could be used as a prognostic indicator for postoperative patients with NSCLC and heparanase might be a promising molecular target for treatment of NSCLC.

Clinical significance of telomerase activity in lung cancer

To evaluate the clinical significance of telomerase activity in lung cancer and to investigate the possibility of telomerase as cancer marker for lung cancer. The activity of telomerase was investigated by TRAP-PCR-ELISA in lung cancer tissues and corresponding adjacent noncancerous tissues obtained from resected specimens of 48 patients with lung cancer and 42 specimens of benign pulmonary lesions were examined simultaneously. Telomerase activity was detected in 42 (87.5%) of the 48 tumors and only 4 (8.3%) of the 48 adjacent noncancerous lung tissue samples (Chi-Square=13.029, P < 0.01), but in none of 42 specimens of benign pulmonary lesions (Chi-Square=14.016, P < 0.01). Correlation with pathological parameters showed that the expression of telomerase activity was associated with lymph node metastasis (93.5% vs 76.4% , Chi-Square=63.511, P < 0.01), but not with histological type, location and differenciated grade of tumor. Telomerase activation correlates with the carcinogenesis and aggressiveness of lung cancer. Telomerase might be one of the important diagnostic and prognostic factors in patients with lung cancer.

The expression of Cx43 protein in non-small cell lung cancer tissues and its clinical significance

To investigate the expression feature of Cx43 protein in non-small cell lung cancer and its clinical significance. Paraffin embedded tissues from 65 patients with primary non-small cell lung cancer and 20 adjacent non-cancerous lung tissues were investigated for expression of Cx43 protein by immunohistochemistry (ABC method). The relationship between Cx43 protein expression and clinicopathological characteristics of lung cancer was analyzed. Cx43 protein was positively expressed in 24 out of 65 lung cancer tissues (36.92%), and in all 20 adjacent non-cancerous lung tissues. There was a significant difference in Cx43 expression between stage I-II and stage III-IV groups ( P < 0.05), as well as between moderate-well and poor differentiation groups ( P < 0.01). Primary lung cancer tissues with lymph node metastasis showed lower expression of Cx43 than those without lymph node metastasis ( P < 0.01). The positive rate of Cx43 expression was not related to histological classification ( P > 0.05). Cx43 expression might play a role in the genesis, development and metastasis of lung cancer. It may be used to judge the biological behavior of lung cancer.

Gene Expression Profiling of Non-Small Cell Lung Cancer.

cDNA microarray provided a powerful alternative, with an unprecedented view scope, in monitoring gene expression levels, and led to the discovery of regulatory pathways involved in complicated biological processes. This study was performed to gain better understanding of the molecular mechanisms underlying the carcinogenesis and progression of lung cancer. Using a cDNA microarray, representing 4, 600 cDNA clusters, we studied the expression profiles in 10 non-small cell lung cancer (NSCLC) samples and the adjacent noncancerous lung tissues form the same patients. The alterations in the levels of gene expression were confirmed by reverse-transcription PCR in 10 randomly selected genes. Genes that were differently expressed in the cancerous and noncancerous tissues were identified. One hundred and nine genes (of which 68 were known) and 69 cDNAs (of which 32 were known) were up- and down-regulated in>70% of the NSCLC samples, respectively. In the cancerous tissues, the genes related to the cell cycle, metabolism, cell structure and signal transduction, were mostly up-regulated. Furthermore, we identified a few putative tumor suppressor genes that had previously been proposed by other workers. S: These results provide, not only a new molecular basis for understanding the biological properties of NSCLC, but also useful resources for the future development of diagnostic markers and therapeutic targets for NSCLC.

Study on clonal expression of T cells bearing dominant TCR Vβ in non-small-cell lung cancers

To investigate the distribution of TCR Vβ subfamily T clonal cells in peripheral blood lymphocytes (PBL), tumor infiltrating lymphocytes (TIL) and lymphocytes in non-cancerous lung tissues of patients with non-small-cell lung cancer (NSCLC) and to see the inclination of the T cell antigen receptor (TCR) Vβ subfamilies' expression. Complimentarily determining region 3 (CDR3) of TCR 24 variable region genes was analyzed in PBL, TIL and lymphocytes in non cancerous lung tissues from 24 NSCLC cases with reverse transcriptase-polymerase chain reaction (RT-PCR) and gene scan techniques to identify the distribution and clonality of TCR Vβ subfamily T cells. Only a portion of Vβ T cells were found in patients with NSCLC, whereas 24 TCR Vβ subfamily T cells were detected in 10 healthy controls. Vβ5 subfamily was expressed mostly in TIL and the frequency of Vβ5 in TIL (6/18, 33.3%) was much higher than that of PBL (1/24, 4.2%) and T cells infiltrating non-cancerous lung tissues (0/12) (P < 0.05). Oligoc lonal T cells were found in 2 cases with Vβ5 subfamily and polyclonal T cells in 4 cases. There are dominant and clonal TCR Vβ subfamilies expressed in TIL of NSCLC patients, which may be the tumor associated antigens (TAA) specific.

A study on the relationship between E-cadherin, β-catenin expression and metastasis and prognosis in non-small cell lung cancer

To investigate the roles of E-cadherin and β-catenin genes in metastasis and prognosis of non-small cell lung cancer. The expression of E-cadherin and β-catenin were detected in 112 non-small cell lung cancer tissues and 30 benign pulmonary lesion tissues by immunohistochemical method (LSAB method). The expression of E-cadherin and β-catenin in lung cancer tissues was significantly lower than that in adjacent non-cancerous lung tissues and benign pulmonary tissues (P < 0.01); The expression of E-cadherin and β-catenin in lung cancer tissues with lymph node and/or distant metastasis was significantly lower than that in those without lymph node and distant metastasis (P < 0.01); The 5-year survival rate in patients with low E-cadherin expression and low β-catenin expression (6.78%) was remarkably lower than that in cases with high expression (6.78% vs 35.85% and 3.78% vs 37.29%) (P < 0.01). Underexpressions of E-cadherin and β-catenin are very common in non-small cell lung cancer, and it may play an important role in the progression and metastasis of lung cancer. Detection of expression of E-cadherin and β-catenin in lung cancer might be helpful to predict prognosis.

Functional cloning, sorting, and expression profiling of nucleic acid-binding proteins.

A major challenge in the post-sequencing era is to elucidate the activity and biological function of genes that reside in the human genome. An important subset includes genes that encode proteins that regulate gene expression or maintain the structural integrity of the genome. Using a novel oligonucleotide-binding substrate as bait, we show the feasibility of a modified functional expression-cloning strategy to identify human cDNAs that encode a spectrum of nucleic acid-binding proteins (NBPs). Approximately 170 cDNAs were identified from screening phage libraries derived from a human colorectal adenocarcinoma cell line and from noncancerous fetal lung tissue. Sequence analysis confirmed that virtually every clone contained a known DNA- or RNA-binding motif. We also report on a complementary sorting strategy that, in the absence of subcloning and protein purification, can distinguish different classes of NBPs according to their particular binding properties. To extend our functional annotation of NBPs, we have used GeneChip expression profiling of 14 different breast-derived cell lines to examine the relative transcriptional activity of genes identified in our screen and cluster analysis to discover other genes that have similar expression patterns. Finally, we present strategies to analyze the upstream regulatory region of each gene within a cluster group and select unique combinations of transcription factor binding sites that may be responsible for dictating the observed synexpression.

Clinicopathologic significance of telomerase activity and hTERT mRNA expression in non-small cell lung cancer

Telomerase has been reported to be a novel diagnostic marker for malignant diseases and has been recently proven to be composed of three main components, hTR (human telomerase RNA component), TP1 (telomerase-associated protein 1) and hTERT (human telomerase reverse transcriptase), the last of which plays a key role in telomerase activation. In the present study, quantitative levels of telomerase activity and hTERT gene mRNA ( hTERT) expression were analyzed in cancerous and non-cancerous lung tissues of 62 lung cancer patients by telomeric repeat amplification protocol and reverse transcription-PCR, respectively. The telomerase expression levels of each group of tissue samples were compared with clinicopathologic variables. Telomerase activity and hTERT were detected in cancerous tissues (75.8 and 75.8%, respectively), while these parameters were not observed in any non-cancerous tissues. In quantitative assessment of telomerase expression, both telomerase activity and hTERT were significantly correlated with lymph node metastasis (N0 vs. N1+2, P<0.05). Telomerase activity also correlated with tumor cell differentiation and stage classification ( P<0.05), but did not correlate with other clinicopathologic variables. The disease-free survival in patients with lung cancer demonstrated that patients with hTERT-positive tumor survived for a significantly shorter period than those with hTERT-negative tumor ( P=0.0334). Since hTERT levels are correlated with N factor which represents the true aggressiveness of patients’ disease concerning the evaluation of clinical outcome, hTERT was found to be one of the important markers revealing biological malignant potentials for lung cancer.

COMPARISON OF C-MYC EXPRESSION IN NORMAL HUMAN BRONCHIAL EPITHELIAL CELLS AND LUNG CANCER CELL LINES USING A QUANTITATIVE FLUORESCENCE-BASED REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION METHOD

Mutations, either heritable or induced,can modulate the expression of oncogenes and/or tumor suppressor genes and thereby alter the biology of the affected cells. Over expression of the c-myc oncogene appears to contribute to the development of small cell lung cancer, and has been used as an indicator of poor prognosis for this tumor type. However, neither the variability of c-myc expression in noncancerous human lung tissue retrieved from different donors nor the level of c-myc expression in various lung tumor types has been adequately studied. The authors havequantified and compared the expression of c-myc in normal human bronchial epithelial (NHBE)cells from several noncancerous lungs and in transformed human lung cell lines. c-myc mRNA levels were determined through the annealing of a gene-specific probe to a target cDNA copy of the mRNA. The fluorescence signal liberated by the 5-3 exonuclease-induced release of the reporter dyefrom theannealed probeduring the extension cycle of polymerasechain reaction (PCR) was quantified spectrophotometrically. While NHBE cells from four donors expressed comparable levels of c-myc mRNA, the c-myc mRNA levels in large cell lung carcinoma (LCLC) and small cell lung carcinoma (SCLC) cell lines were 2.4- and 5.0-fold greater than in NHBE cells, respectively. In contrast, the lung adenocarcinoma and adenosquamous carcinoma cell lines examined did not exhibit an increase in c-myc expression relative to the NHBE cells. In summary, this reverse/transcriptase polymerasechain reaction method readily quantifies and distinguishes the level of c-myc mRNA in NHBE and human lung cancer cells. This technology may serve to characterize oncogene-related changes during tumorigenesis, to identify genetically predisposed individuals, and to allow for earlier diagnosis and treatment of lung cancer.

Expression of Transforming Growth Factor-alpha and Transforming Growth Factor-beta In Human Primary Lung Cancers

Background: Transforming growth factor- alpha(TGF-) may play important roles in carcinogenesis, tumor growth, and angiogenesis. Transforming growth factor-beta(TGF-) are known to be involved in cell-cycle control and regeneration. TGF- positively acts on growth control of many epithelial cells in contrast to the negative role of TGF-. Method: To evaluate the possible role of TGF- and TGF- in human primary lung cancers, the expression of TGF- and TGF- were immmunohistochemically investigated in tissue sections from forty seven cases with lung cancers and ten cases with non-cancerous lung tissues. Recombinant cloned monoclonal antibody of TGF- and neutralizing antibody of TGF- were employed as primary antibodies after dewaxing the formalin-fixed, paraffinized tissue sections. Results: TGF- was expressed in the cytoplasms of tumor cells in thirty five cases of forty seven(74.5%) primary lung cancers, whereas the control expressed in two of ten brochial epithelial cells. The expression of TGF- was disclosed in four cases of eleven(36.4 %) small cell carcinomas and thirty one cases of thirty six(86.1%) non-small cell carcinomas of the lung. Expressions of TGF- was discernible in bronchial epithelium in eight of ten non-cancerous lung tissues. The expression of TGF- was noted in the cytoplasms of tumor cells in eight cases of forty seven(17.0%) primary lung cancers. The expression of TGF- disclosed in two cases of eleven(18.2%) small cell carcinomas and six cases of thirty six(16.7%) non- small cell carcinomas of the lung. Conclusion: These findings suggest that up-regulation of TGF- and down-regulation of TGF- are involved during development and growth of primary lung cancers.

Higher Expression of Topoisomerase II in Lung Cancers Than Normal Lung Tissues: Different Expression Pattern from Topoisomerase I

To examine the expression of topoisomerase I and topoisomerase II in primary lung cancer specimens at mRNA level, we carried out Northern blot analysis. As for topoisomerase I expression, there was no remarkable difference between lung cancer specimens and non-cancerous lung tissues. On the other hand, we could detect topoisomerase II mRNA in almost all lung cancer specimens, but not in non-cancerous tissues. By Southern blot analysis, we could not detect large deletion nor rearrangement in DNA level. These results suggest that the expression of topoisomerase II is highly increased in lung cancer at mRNA level and drugs against topoisomerase II might be more tumor-specific than those against topoisomerase I.

ESTIMATING RISKS TO PREDICT MORBIDITY AND MORTALITY AFTER PULMONARY RESECTION FOR LUNG CANCER WITH PULMONARY EMPHYSEMA

Preoperative risks were estimated in resected cases of lung cancer associated with pulmonary emphysema. Subjects were 42 out of 136 cases undergone pulmonary resection for a primary lung cancer, which were selected by satisfying the following conditions, as well as by referring pathological findings of resected non-cancerous lung tissues: preoperative FEV1.0, less than 70%; RV/TLC, more than 35%; and improvement rate of forced expiratory volume in one second after administration of bronchodilator, less than 500ml. Critical postopertive complications were observed in 15 patients (35.7%). There were 5 operative deaths and another 5 deaths in the hospital among the 15 patients. Predicted-FEVI1.0/VBSA, %DLco/VA and CT-findings are more important predictors of morbidity and mortality. When the cases had at least 2 out of 3 predictors, namely, P-FEV1.0/BSA of less than 0.85l/m2, %DLco/VA of less than 60% and CT-grade III, postoperative complications occurred in an extremely high rate. Combination of those predictors should be reliabloe estimation of preoperative risk foctors for patients with lung cancer associated with pulmonary emphysema.