Diagnostic value of α-enolase expression and serum α-enolase autoantibody levels in lung cancer / Valor diagnóstico da expressão de α-enolase e dos níveis séricos de autoanticorpos contra α-enolase no câncer de pulmão

ABSTRACT Objective: To investigate the diagnostic value of α-enolase (ENO1) and serum ENO1 autoantibody levels in lung cancer. Methods: Immunohistochemistry staining and ELISA were performed to detect ENO1 expression in lung tissue and serum ENO1 autoantibody levels, respectively. Results: The expression of ENO1 was higher in lung cancer tissues than in benign lung disease tissues (p < 0.001). The proportion of lung cancer samples expressing ENO1 was not significantly different among the various pathological classification groups. The proportion of samples expressing ENO1 was higher in lung cancer patients in stages I/II than in those in stages III/IV (χ2 = 5.445; p = 0.018). The expression of ENO1 in lung cancer tissues was not associated with age, gender, or smoking history. Serum ENO1 antibody levels were significantly higher in the lung cancer group than in the benign lung disease and control groups (p < 0.001). The differences among the pathological classification groups were not statistically significant. Serum ENO1 antibody levels were also in lung cancer patients in stages I/II than in those in stages III/IV (p < 0.01). Serum ENO1 antibody levels were not associated with age, gender, or smoking history in lung cancer patients. The ROC curve representing the diagnosis of lung cancer based on ENO1 antibody levels had an area under the curve of 0.806. Conclusions: Our results suggest that high levels of ENO1 are associated with the clinical stage of lung cancer and that ENO1 expression and its serum autoantibody levels show diagnostic value in lung cancer.

RESUMO Objetivo: Investigar o valor diagnóstico da α-enolase (ENO1) e dos níveis séricos de autoanticorpos contra ENO1 no câncer de pulmão. Métodos: Marcação imuno-histoquímica e ELISA foram realizados para detectar a expressão de ENO1 no tecido pulmonar e os níveis séricos de autoanticorpos contra ENO1, respectivamente. Resultados: A expressão de ENO1 foi maior nos tecidos de câncer de pulmão que nos tecidos de doença pulmonar benigna (p < 0,001). Não houve diferença significativa entre os diversos grupos de classificação patológica quanto à proporção de amostras de câncer de pulmão que expressaram ENO1. A proporção de amostras que expressaram ENO1 foi maior nos pacientes com câncer de pulmão nos estágios I/II que naqueles com câncer de pulmão nos estágios III/IV (χ2 = 5,445; p = 0,018). Não houve relação entre a expressão de ENO1 em tecidos de câncer de pulmão e idade, sexo ou histórico de tabagismo. Os níveis séricos de anticorpos contra ENO1 foram significativamente maiores no grupo câncer de pulmão que nos grupos doença pulmonar benigna e controle (p < 0,001). As diferenças entre os grupos de classificação patológica não foram estatisticamente significativas. Os níveis séricos de anticorpos contra ENO1 foram também significativamente maiores nos pacientes com câncer de pulmão nos estágios I/II que naqueles com câncer de pulmão nos estágios III/IV (p < 0,01). Nos pacientes com câncer de pulmão, não houve relação entre os níveis séricos de anticorpos contra ENO1 e idade, sexo ou histórico de tabagismo. A curva ROC do diagnóstico de câncer de pulmão baseado nos níveis de anticorpos contra ENO1 apresentou área sob a curva = 0,806. Conclusões: Nossos resultados sugerem que há relação entre níveis elevados de ENO1 e o estágio clínico do câncer de pulmão e que a expressão de ENO1 e os níveis séricos de autoanticorpos contra ENO1 têm valor diagnóstico no câncer de pulmão.

Hyaluronidases and hyaluronan synthases expression is inversely correlated with malignancy in lung/bronchial pre-neoplastic and neoplastic lesions, affecting prognosis

We collected a series of 136 lung/bronchial and 56 matched lung parenchyma tissue samples from patients who underwent lung/bronchial biopsies and presented invasive carcinoma after lung surgery. The lung/bronchial samples included basal cell hyperplasia, squamous metaplasia, moderate dysplasia, adenomatous hyperplasia, severe dysplasia, squamous cell carcinoma and adenocarcinoma. Matched lung parenchyma tissue samples included 25 squamous cell carcinomas and 31 adenocarcinomas. Immunohistochemistry was performed to analyze for the distribution of hyaluronidase (Hyal)-1 and −3, and hyaluronan synthases (HAS)-1, −2, and −3. Hyal-1 showed significantly higher expression in basal cell hyperplasia than in moderate dysplasia (P=0.01), atypical adenomatous hyperplasia (P=0.0001), or severe dysplasia (P=0.03). Lower expression of Hyal-3 was found in atypical adenomatous hyperplasia than in basal cell hyperplasia (P=0.01) or moderate dysplasia (P=0.02). HAS-2 was significantly higher in severe dysplasia (P=0.002) and in squamous metaplasia (P=0.04) compared with basal cell hyperplasia. HAS-3 was significantly expressed in basal cell hyperplasia compared with atypical adenomatous hyperplasia (P=0.05) and severe dysplasia (P=0.02). Lower expression of HAS-3 was found in severe dysplasia compared with squamous metaplasia (P=0.01) and moderate dysplasia (P=0.01). Epithelial Hyal-1 and −3 and HAS-1, −2, and −3 expressions were significantly higher in pre-neoplastic lesions than in neoplastic lesions. Comparative Cox multivariate analysis controlled by N stage and histologic tumor type showed that patients with high HAS-3 expression in pre-neoplastic cells obtained by lung/bronchial biopsy presented a significantly higher risk of death (HR=1.19; P=0.04). We concluded that localization of Hyal and HAS in lung/bronchial pre-neoplastic and neoplastic lesions was inversely related to malignancy, which implied that visualizing these factors could be a useful diagnostic procedure for suspected lung cancer. Finalizing this conclusion will require a wider study in a randomized and prospective trial.

A socialização da medicina na era do adhemarismo / The socialization of medicine in the era of São Paulo Governor Adhemar de Barros

A análise observa as conexões entre o processo de profissionalização dos médicos paulistas e políticas de saúde do governo estadual de Adhemar de Barros em São Paulo (1947-1951), em meio a amplas mudanças na área de saúde denominadas pelos médicos paulistas “socialização da medicina”. Reconhecemos aspectos ambivalentes para o profissionalismo médico diante desse governo populista, como: a luta médica pela equiparação ante os advogados servidores públicos estaduais; a criação de uma secretaria de saúde estadual; e certos elos contraditórios entre a área de saúde adhemarista e a ideologia e a organização profissionais da medicina paulista. Nesse particular, o artigo aprofunda a análise de manifestações ideológicas de importantes lideranças médicas paulistas.

The article analyzes how the process of the professionalization of physicians in São Paulo related to healthcare policy under the administration of São Paulo governor Adhemar de Barros (1947-1951) during a period of broad change in the realm of health known by São Paulo physicians as the “socialization of medicine.” Medical professionalism confronted certain ambivalences under this populist administration, including doctors’ struggle to achieve pay equal to that of state public attorneys; the establishment of a state health department; and some contradictory ties between the area of health under Adhemar and the professional ideology and organization of medicine in São Paulo. The article undertakes a more in-depth analysis of the ideological manifestations of important leaders in the state’s medical community.

Predictive factors of alcohol and tobacco use in adolescents / Fatores preditivos do uso de álcool e tabaco em adolescentes / Factores predictivos del uso de alcohol y tabaco en los adolescentes

OBJECTIVES: to analyze the effect of self-esteem, assertiveness, self-efficacy and resiliency on alcohol and tobacco consumption in adolescents. METHOD: a descriptive and correlational study was undertaken with 575 adolescents in 2010. The Self-Esteem Scale, the Situational Confidence Scale, the Assertiveness Questionnaire and the Resiliency Scale were used. RESULTS: the adjustment of the logistic regression model, considering age, sex, self-esteem, assertiveness, self-efficacy and resiliency, demonstrates significance in the consumption of alcohol and tobacco. Age, resiliency and assertiveness predict alcohol consumption in the lifetime and assertiveness predicts alcohol consumption in the last year. Similarly, age and sex predict tobacco consumption in the lifetime and age in the last year. CONCLUSION: this study can offer important information to plan nursing interventions involving adolescent alcohol and tobacco users. .

OBJETIVOS: analisar o efeito da autoestima, assertividade, autoeficácia e resiliência sobre o consumo de álcool e tabaco em adolescentes. MÉTODO: estudo descritivo correlacional com 575 adolescentes, realizado no ano 2010. Foram utilizadas a Escala de Autoestima, o Questionário de Confiança Situacional, o Questionário de Assertividade e a Escala de Resiliência. RESULTADOS: o ajuste do modelo de regressão logística, considerando a idade, sexo, autoestima, assertividade, autoeficácia e resiliência foi significante em relação ao consumo de álcool e tabaco. A idade, resiliência e assertividade foram preditores do consumo de álcool em algum momento na vida e a idade e a assertividade foram preditores no último ano. Para o consumo de tabaco, a idade e o sexo foram preditores em algum momento na vida e a idade no último ano. CONCLUSÃO: este estudo pode proporcionar informações importantes para o planejamento de intervenções de enfermagem em adolescentes usuários de álcool e tabaco .

OBJETIVOS: analizar el efecto de la autoestima, asertividad, autoeficacia y resiliencia sobre el consumo de alcohol y tabaco en adolescentes. MÉTODO: descritivo correlacional con 575 adolescentes, en 2010. Se utilizaron la Escala de Autoestima, el Cuestionario de Confianza Situacional, el Cuestionario de Asertividad y la Escala de Resiliencia. RESULTADOS: el ajuste del modelo de regresión logística, considerando la edad, sexo, autoestima, asertividad, autoeficacia y resiliencia, muestra significancia en el consumo de alcohol y tabaco. La edad, resiliencia y asertividad predicen el consumo de alcohol alguna vez en la vida y la edad y asertividad en el último año. De la misma forma la edad y sexo predicen el consumo de tabaco alguna vez en la vida y la edad en el último año. CONCLUSIÓN: este estudio puede proporcionar información importante para la planificación de intervenciones en enfermería de los adolecentes usuarios de alcohol y tabaco. .

Telomerase Activity and the Risk of Lung Cancer

Telomerase play a key role in the maintenance of telomere length and chromosome integrity. We have evaluated the association between telomerase activity and the risk of lung cancer in peripheral blood. Telomerase activity in peripheral blood mononuclear cells was measured by a PCR-designed telomeric repeat amplification protocol in 63 lung cancer patients and 190 healthy controls that were matched for age, gender, and smoking status. Telomerase activity was significantly lower in the lung cancer patients than in controls (mean +/- standard deviation; 1.32 +/- 1.65 vs 2.60 +/- 3.09, P < 1 x 10(-4)). When telomerase activity was categorized into quartiles based on telomerase activity in the controls, the risk of lung cancer increased as telomerase activity reduced (Ptrend = 1 x 10(-4)). Moreover, when the subjects were categorized based on the median value of telomerase activity, subjects with low telomerase activity were at a significantly increased risk of lung cancer compared to subjects with high telomerase activity (adjusted odds ratio = 3.05, 95% confidence interval = 1.60-5.82, P = 7 x 10-4). These findings suggest that telomerase activity may affect telomere maintenance, thereby contributing to susceptibility to lung cancer.

Cassia alata leaf extract induces cytotoxicity in A549 lung cancer cells via a mechanism that is caspase 8 dependent / El extracto de la hoja de cassia alata induce citotoxicidad en las células A549 del cáncer pulmonar humano a través de un mecanismo dependiente de la caspasa 8

OBJECTIVE: To evaluate the cytotoxic effect of a hexane extract of Cassia alata leaves in A549 lung cancer cells. METHOD: Parental A549 lung cancer cells were exposed to various concentrations (100"180 µg/ml) of Cassia alata leaf extract for 24 hours. Following treatment, the cells were evaluated using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay to determine the cytotoxic effect of the extract. Caspase 8, 3 and 9 negative A549 cells were also prepared using lentiviral based shRNA knockdown of the caspase 8, 3 and 9 genes, respectively. The cytotoxic effect of Cassia alata leaf extract was then evaluated in these knockdown cells using the MTT assay. Chemical analysis was performed on the extract using high performance liquid chromatography (HPLC). RESULTS: Cassia alata extract was cytotoxic in parental and caspase-9 negative, but not caspase 3 and 8 negative A549 cells. The IC50 values were 143 µg/ml and 145 µg/ml in parental and caspase 9 negative A549 cells respectively. The flavanoid kaempferol was identified as a constituent of Cassia alata leaf extract. CONCLUSIONS: Cassia alata produces cytotoxicity in A549 cancer cells that is mediated by caspase 8 activation. This effect may be attributable to kaempferol.

OBJETIVO: Evaluar el efecto citotóxico de un extracto de hexano de hojas de Cassia alata en las células A549 del cáncer pulmonar. MÉTODO: Células A549 parentales del cáncer pulmonar fueron expuestas a varias concentraciones (100-180 µg/ml) de un extracto de la hoja de Cassia alatadurante 24 horas. Tras el tratamiento, las células fueron evaluadas usando el ensayo de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolio (MTT) a fin de determinar el efecto citotóxico del extracto. También se prepararon células A549 negativas caspasa 8, 3 y 9 mediante silenciamiento génico vía ARN (shRNA knockdown) de los genes de las caspasas 8, 3 y 9 respectivamente, sobre la base de la inserción de vectores lentivirales. Entonces, usando un ensayo MTT se procedió a evaluar el efecto citotóxico del extracto de hojas de Cassia alataen éstas células genéticamente modificadas. Se realizó un análisis químico del extracto utilizando cromatografía líquida de alta eficacia. (HPLC). RESULTADOS: El extracto de Cassia alata resultó ser citotóxico en las células A549 negativas parentales y caspasa 9, pero no en las negativas caspasa 3 y 8. Los valores de IC50 fueron 143 µg/ml y 145 µg/ml en las células A549 negativas parentales y caspasa 9 respectivamente. El flavonol kaempferol fue identificado como un constituyente del extracto de las hojas de Cassia alata. CONCLUSIONES: La Cassia alata produce citotoxicidad en las células cancerosas A549, mediada por la activación de la caspasa 8. Este efecto puede ser atribuido al kaempferol.

Nuclear factor E2-related factor 2 Dependent Overexpression of Sulfiredoxin and Peroxiredoxin III in Human Lung Cancer

BACKGROUND/AIMS: Oxidative stress results in protein oxidation and is implicated in carcinogenesis. Sulfiredoxin (Srx) is responsible for the enzymatic reversal of inactivated peroxiredoxin (Prx). Nuclear factor E2-related factor 2 (Nrf2) binds to antioxidant responsive elements and upregulates the expression of Srx and Prx during oxidative stress. We aimed to elucidate the biological functions and potential roles of Srx in lung cancer. METHODS: To study the roles of Srx and Prx III in lung cancer, we compared the protein levels of Nrf2, Prxs, thioredoxin, and Srx in 40 surgically resected human lung cancer tissues using immunoblot and immunohistochemical analyses. Transforming growth factor-beta1, tumor necrosis factor-alpha, and camptothecin treatment were used to examine Prx III inactivation in Mv1Lu mink lung epithelial cells and A549 lung cancer cells. RESULTS: Prx I and Prx III proteins were markedly overexpressed in lung cancer tissues. A significant increase in the oxidized form of a cysteine sulfhydryl at the catalytic site of Prxs was found in carcinogenic lung tissue compared to normal lung tissue. Densitometric analyses of immunoblot data revealed significant Srx expression, which was higher in squamous cell carcinoma tissue (60%, 12/20) than in adenocarcinoma (20%, 4/20). Also, Nrf2 was present in the nuclear compartment of cancer cells. CONCLUSIONS: Srx and Prx III proteins were markedly overexpressed in human squamous cell carcinoma, suggesting that these proteins may play a protective role against oxidative injury and compensate for the high rate of mitochondrial metabolism in lung cancer.

Implication of leucyl-tRNA synthetase 1 (LARS1) over-expression in growth and migration of lung cancer cells detected by siRNA targeted knock-down analysis

Molecular mechanism of lung carcinogenesis and its aggressive nature is still largely elusive. To uncover the biomarkers related with tumorigenesis and behavior of lung cancer, we screened novel differentially expressed genes (DEG) in A549 lung cancer cell line by comparison with CCD-25Lu, normal pulmonary epithelial cell line, using annealing control primer(ACP)- based GeneFishing system. Of the DEGs, over-expression of leucyl-tRNA synthetase 1 (LARS1) was prominent and this up-regulation was confirmed by immunoblotting and real-time quantitative RT-PCR analysis. In addition to A549 cell line, primary lung cancer tissues also expressed higher level of LARS1 mRNA than their normal counter tissues. To explore the oncogenic potential of LARS1 over-expression in lung cancer, we knocked-down LARS1 by treating siRNA and observed the tumor behavior. LARS1 knock-down cells showed reduced ability to migrate through transwell membrane and to form colonies in both soft agar and culture plate. Taken together, these findings suggest that LARS1 may play roles in migration and growth of lung cancer cells, which suggest its potential implication in lung tumorigenesis.

Role of cyclooxygenase-2 in tumor progression and immune regulation in lung cancer.

Lung cancer is the leading cause of cancer death all over the world. The low 5-year survival rate (under 15%) has changed minimally in the last 25 years. Amongst different types of lung cancers, non-small cell lung carcinoma (NSCLC) types account 25-40%. To improve the survival of lung cancer patients, new therapeutic strategies are needed. The search for improved therapies has led to the investigation of agents that target novel pathways involved in tumor proliferation, invasion, survival and immune regulation. Cyclooxygenase-2 (COX-2) is one of the novel targets under evaluation for NSCLC therapy and chemoprevention. Although multiple genetic alterations are necessary for lung cancer invasion and metastasis, COX-2 may act as central element in orchestring these processes. COX-2 plays an important role in all aspects of tumor development and growth. It also plays a pivotal role in regulation of cytokines and immune responses in NSCLC patients. In this article, we review the experimental and clinical evidences on the possible link between COX and NSCLC.

Expression and significance of cyclooxygenase 2 gene in lung cancer / 华中科技大学学报（医学）（英德文版）

To study the expression of cyclooxygenase 2 (COX-2) gene and its relationship with clinicopathological characteristics of lung cancer, expression of the COX-2 mRNA was evaluated by reverse transcription polymerase chain reaction (RT-PCR) in cancerous tissues and paired adjacent non-cancerous tissues from 56 patients and benign lesions from 12 patients. Our results showed that expression of COX-2 gene was detected in a significantly greater proportion of cancerous tissues (60.7%) than adjacent noncancerous tissues (10.7%, P0.05). The up-regulation of COX-2 gene in lung cancer tissues especially in adenocarcinoma suggested that COX-2 may play a role in the lung carcinogenesis and COX-2 gene may serve as a potential therapeutic target in lung cancer.

Hydrogen peroxide mediates doxorubicin-induced transglutaminase 2 expression in PC-14 human lung cancer cell line

Increased expression of Transglutaminases 2 (TGase 2, TGase C) was observed in PC-14 human lung cancer cells in association with doxorubicin resistance and the reduction of the enzyme expression was correlated with the increasing cytotoxicity of the drug (Han and Park, 1999). Hydrogen peroxide was suggested to be a key mediator for doxorubicin-induced DNA fragmentation leading to apoptosis. A possible role of hydrogen peroxide as a putative mediator of TGase 2 expression in the doxorubicin sensitive PC-14 cells was examined. TGase 2 expression was increased in PC-14 cells treated with doxorubicin in a dose-dependent manner resulting in the concomitant increase of reactive oxygen species. The rise of TGase 2 expression by doxorubicin treatment was inhibited by N-acetylcysteine or glutathione treatment, while direct addition of hydrogen peroxide to PC-14 cells induced TGase 2 expression. These results suggest that generation of hydrogen peroxide induced by doxorubicin treatment is one of the key factors in an enhancement of TGase 2 expression in PC-14 cells.

Serum adenosine deaminase in pulmonary tuberculosis, malignancy and non-tubercular respiratory diseases.

The study has been conducted to find out the serum ADA levels in 120 patients with various pulmonary diseases which included patients with tubercular pleural effusion (n = 86), lung cancer (n = 10) and patients with non-tubercular pulmonary diseases like pneumonia, etc (n = 24). Twenty healthy individuals served as control subjects. The mean (+/- SD) of ADA activity was 23.38 (4.47), 7.29 (1.08), 12.71 (1.95) and 2.23 (1.00) units/litre in tuberculosis, malignancy, non-tubercular pulmonary diseases and healthy controls respectively with significant difference between each other (P less than 0.001). Patients with tuberculosis (100%) fall in 97% sensitivity range with a lower cut off limit at 17 units/litre ADA activity, while for malignancy and non-tubercular respiratory diseases, the sensitivity was 90% and 83% respectively. Within the sensitivity limits, the serum ADA activity can be used for the differential diagnosis of pulmonary diseases.

Elucidation of sialyltransferase as a tumour marker.

The report describes results of separation of sialyltransferase isoenzymes by electrofocusing plasma from healthy volunteers and patients having different types of malignant tumour. Extensive modification of the technique was adopted in determining enzyme activity, such as elution of gel strips with the buffer pH corresponding to the gel focusing point; assessment of the effect of different pH on endogenous incorporation of radioactivity to desialated fetuin; and quantitative analysis of protein present in each gel band for calculation of enzyme activity. Plasma from normal individuals showed the existence of 5 sialyltransferase isoenzymes at pI 4.8, 5.5, 6.3, 6.8 and 7.5. There were higher isoenzyme activities in plasma samples from patients afflicted with malignancy of lungs and colon in comparison to normal pattern. Endometrial and breast cancer patients also showed elevated levels of the enzyme which could be controlled by surgery and combined therapies with cytotoxic drugs and radiation, respectively. The observations suggest the potential use of sialyltransferase as a tool for tumour diagnosis, and are discussed in relation to prognosis of the disease in the course of therapy.