Diagnostic value of plasma expression of microRNAs complementary to Drosha and Dicer in lung cancer patients.

OBJECTIVE: Lung cancer (LC) is diagnosed mostly in advanced, non-operable stage, with poor prognosis. The analysis of microRNAs may be a useful tool for early and non-invasive detection of cancer. Dicer and Drosha are enzymes with an essential role for microRNA biogenesis. The aim of our study was to analyze the expression of miRNA-27a-3p, miRNA-31, miRNA-182, miRNA-195 with the ability to reciprocal regulation of Dicer and Drosha expression in lung cancer patients. PATIENTS AND METHODS: The relative expression of microRNAs was detected by qPCR in plasma of 160 LC patients. The U-Mann Whitney test was used to compare the relative expression between particular groups of lung cancer patients and healthy individuals. The diagnostic value of microRNAs examination was analyzed using a receiver operating curve. RESULTS: We demonstrated that the plasma levels of miRNA-27, miRNA-31 and miRNA-182 were significantly higher and miRNA-195 significantly lower in the whole group of LC patients and in patients with early stages of NSCLC, in comparison with healthy donors. ROC analysis showed that four studied microRNAs have a potential diagnostic value for early stages of NSCLC with AUC=0.95 for miRNA-27a (94% sensitivity and 81% specificity, p=0.0001), 0.71 for miRNA-31 (73% sensitivity and 61% specificity, p=0.001) 0.77 for miRNA-182 (70% sensitivity and 79% specificity, p=0.0001) and 0.82 for miRNA-195 (74% sensitivity and 80% specificity, p=0.0001). CONCLUSIONS: We have proved that the expression of miRNA-27a-3p, miRNA-31, miRNA-182, and miRNA-195 in patients with LC is different from the expression of these molecules in healthy people. The examination of these microRNAs in plasma could be used in non-invasive lung cancer diagnosis.

Serum from patients with chronic obstructive pulmonary disease promotes proangiogenic behavior of the vascular endothelium.

OBJECTIVE: It has been documented that COPD is a risk factor for lung cancer. In COPD patients, changes in lung angiogenesis - a critical process in the development of lung cancer - have been poorly investigated. We aimed to determine whether serum from COPD patients could promote the proangiogenic capabilities of endothelial cells in vitro. PATIENTS AND METHODS: The research was carried out using sera from COPD patients and healthy volunteers, endothelial cells EA.hy926, and bronchial epithelial cells. The concentration of angiogenic molecules was quantified using ELISA tests. The proliferation and migration of EA.hy926 were tested using fluorescence-based methods. Tube formation was analyzed with a commercially available assay. RESULTS: Sera from COPD patients and conditioned media generated by epithelial cells exposed to these sera stimulate proliferation, but not migration, of EA.hy926. This coincided with increased tube formation in both experimental regimens. The sera from COPD patients contained increased levels of CCL2, CCL21, and HGF, whereas the conditioned media generated by epithelial cells treated with these sera exhibited increased levels of CCL2, CCL21, CXCL8, FGF, and sICAM-1. The concentration of angiogenic markers in the sera and conditioned media, and their effect on the behavior of the endothelium were independent of smoking status (COPD and controls), stage of obstruction, and disease group (COPD). CONCLUSIONS: The increased incidence of lung malignancy in COPD patients may be associated, at least to some extent, with the direct and indirect proangiogenic activity of their sera (via alterations in the secretome of epithelial cells).

Methylation of the DCLK1 promoter region in circulating free DNA and its prognostic value in lung cancer patients

Introduction: The possibility of detection of suppressor genes methylation in circulating free DNA (cf-DNA) of cancer patients and the lack of methylation in healthy individuals makes this epigenetic alternation an ideal diagnostic marker of neoplastic processes. Moreover, hypermethylation in several genes promoter was described as a biomarker of lung cancer. Methylation in the gene encoding doublecortin-like kinase 1 (DCLK1) is observed in patients with colorectal cancer and cholangiocarcinoma. However, there are no studies concerning DCLK1 methylation in lung cancer patients. The aims of the study was to evaluate the frequency of DCLK1 promoter methylation in cf-DNA of lung cancer patients and of healthy persons as well as the usefulness of this test for predicting the lung cancer course. Materials and methods: DCLK1 methylation status was evaluated in DNA isolated from peripheral blood plasma from 65 lung cancer patients and 95 healthy individuals. After DNA bisulfitation, DCLK1 methylation was determined using the qMSP-PCR technique. Moreover, the presence of DCLK1 methylation was correlated with the overall survival (OS) probability of lung cancer patients. Results: DCLK1 promoter methylation was detected in 32 lung cancer patients (49.2 %) and 8 healthy individuals (8.4 %). The methylation of the region before transcription start site (TSS) and the region after TSS of DCLK1 gene was detected in 28 and 11 patients, respectively. In seven cases (10.8 %), the DCLK1 promoter methylation in both regions was reported simultaneously. The methylation was observed slightly frequently in patients with small cell lung cancer (75 % of SCLC patients). The median overall survival of patients with DCLK1 promoter methylation was lower than that of patients without DCLK1 gene modification (p = < 0.001, HR = 4.235). Conclusions: The evaluation of DCLK1 promoter region methylation may be useful in both early diagnosis and prediction of the course of lung cancer (AU)

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Methylation of the DCLK1 promoter region in circulating free DNA and its prognostic value in lung cancer patients.

INTRODUCTION: The possibility of detection of suppressor genes methylation in circulating free DNA (cf-DNA) of cancer patients and the lack of methylation in healthy individuals makes this epigenetic alternation an ideal diagnostic marker of neoplastic processes. Moreover, hypermethylation in several genes promoter was described as a biomarker of lung cancer. Methylation in the gene encoding doublecortin-like kinase 1 (DCLK1) is observed in patients with colorectal cancer and cholangiocarcinoma. However, there are no studies concerning DCLK1 methylation in lung cancer patients. The aims of the study was to evaluate the frequency of DCLK1 promoter methylation in cf-DNA of lung cancer patients and of healthy persons as well as the usefulness of this test for predicting the lung cancer course. MATERIALS AND METHODS: DCLK1 methylation status was evaluated in DNA isolated from peripheral blood plasma from 65 lung cancer patients and 95 healthy individuals. After DNA bisulfitation, DCLK1 methylation was determined using the qMSP-PCR technique. Moreover, the presence of DCLK1 methylation was correlated with the overall survival (OS) probability of lung cancer patients. RESULTS: DCLK1 promoter methylation was detected in 32 lung cancer patients (49.2 %) and 8 healthy individuals (8.4 %). The methylation of the region before transcription start site (TSS) and the region after TSS of DCLK1 gene was detected in 28 and 11 patients, respectively. In seven cases (10.8 %), the DCLK1 promoter methylation in both regions was reported simultaneously. The methylation was observed slightly frequently in patients with small cell lung cancer (75 % of SCLC patients). The median overall survival of patients with DCLK1 promoter methylation was lower than that of patients without DCLK1 gene modification (p = <0.001, HR = 4.235). CONCLUSIONS: The evaluation of DCLK1 promoter region methylation may be useful in both early diagnosis and prediction of the course of lung cancer.

Circulating adipokines and organ involvement in patients with systemic sclerosis.

BACKGROUND: In recent years, mediators synthesized in the adipose tissue, the so-called adipokines, have been reported to play important roles in the pathogenesis of autoimmune rheumatic diseases. OBJECTIVE: To compare serum leptin, adiponectin and resistin levels in patients with systemic sclerosis (SSc) and healthy controls. To find possible relationship between serum levels of adipokines and organ involvement with focus on interstitial lung disease in SSc patients. PATIENTS AND METHODS: Lung involvement was assessed functionally (body plethysmography, diffusing capacity of the lung for carbon monoxide (DLCO) and six-minute walk test) and radiologically (using average disease extent on high resolution computed tomography (HRCT) of the lungs according to the percentage of interstitial changes) in 29 SSc patients. Quantitative sandwich ELISA was used to measure resistin, leptin and adiponectin concentrations in sera of patients and 30 healthy controls. RESULTS: We found no statistically significant differences in serum resistin, leptin and adiponectin levels between SSc patients and the controls. However, serum adiponectin concentrations were significantly lower in active than in inactive patients, they also correlated positively with vital capacity (VC) (p=0.04) and negatively with Valentini activity score (p=0.04). Serum resistin levels were significantly elevated in patients with digital ulcers (p=0.03) and serum concentrations of leptin were associated with the duration of SSc symptoms other than Raynaud's phenomenon (p<0.01) CONCLUSIONS: Serum adiponectin should be further investigated as a candidate for SSc activity marker and resistin may play a role in ulcer development in SSc patients.

Lung impairment in scleroderma.

Scleroderma typically manifests as fibrosis of the skin, but may also involve other organs, particularly the lungs. Interstitial lung disease and functional abnormalities are observed in the majority of patients. The aim of this study was to evaluate radiological changes in the lungs and their correlation with functional disorders in scleroderma patients. The study was conducted in 37 scleroderma patients (F/M-31/6). High resolution computed tomography (HRCT), Warrick score system and spirometry, body plethysmography, and lung diffusion examinations (DLco) were performed. The HRCT showed septal and subpleural lines in 70%, ground-glass opacities in 51%, and honeycomb lungs in 30% of the cases. The DLco values were decreased in 92% of the patients. Total lung capacity (TLC) showed a restrictive pattern in 24% of the patients, and only in 11% of them obstruction predominated. The Warrick score correlated inversely with both DLco (r=0.36; p>0.05). Interstitial lung disease often coexists with scleroderma and is accompanied by functional lung abnormalities.

Concentration of matrix metalloproteinase-9 in serum of patients with chronic obstructive pulmonary disease and a degree of airway obstruction and disease progression.

Chronic obstructive pulmonary disease (COPD) is chronic inflammation leading to irreversible airway obstruction. Previous studies showed increased metalloproteinases (MMP) level, especially MMP-9, as a sign of local inflammation. Up-to-date, only a few studies estimated the MMP-9 serum concentration in COPD with respect to correlation with systemic inflammation. The aim of the present study was to estimate the MMP-9 serum concentration in COPD and to evaluate the correlation between MMP-9 and a degree of airway obstruction in COPD. Twenty three COPD patients and 23 healthy controls were enrolled. In both groups spirometry was performed. MMP-9 concentration in sera taken from both groups was studied using ELISA. We found that COPD patients had increased serum MMP-9 concentration compared with the control group (P=0.0005). In the COPD group, the MMP-9 levels were negatively correlated with FEV1 (P=0.01) and FEV1/FVC (P=0.0002). In conclusion, the results suggest that MMP-9 plays an important role in systemic inflammation in COPD. Higher MMP-9 serum concentration is connected with higher airway obstruction and disease progression.

Tobacco smoking behavior among high school students in Poland.

The aim of the study was to examine Polish students' smoking behavior. A group of 671 students completed a questionnaire about smoking. 53.1% of them tried smoking at least once. The most common age at an attempt to smoke was 18 years. Males tried smoking earlier than females. 23.1% were active smokers during the study. The age of 18 was the most common time when smoking became regular. Males smoked more frequently than females. There were significant differences between the kind of school and the frequency of smoking. Students of smoking parents smoked more frequently than those of non-smoking ones. Most smokers (80%) were aware of detrimental effects of addict and declared a will to quit smoking. We conclude that a substantial percentage of Polish students smoke. Anti-tobacco prevention must be started early, before the age of 18 and continued up to 21.