Wojciech Naumnik

Serum levels of HMGB1, survivin, and VEGF in patients with advanced non-small cell lung cancer during chemotherapy.

Recently, several reports have suggested that HMGB1 (the high-mobility group box-1) plays a key role in tumor angiogenesis through multiple mechanisms, including up-regulation of proangiogenic factors. This study was conducted to investigate the prognostic role and the effects of chemotherapy on serum (ELISA) angiogenic factors: HMGB1, survivin and VEGF (Vascular Endothelial Growth Factor) in patients with advanced stage non-small cell lung cancer (NSCLC). The study entered 40 patients (31 man) and 15 healthy volunteers (control group). Peripheral blood samples were taken before and after four cycles of chemotherapy. The mean serum HMGB1 and VEGF levels were significantly higher in patients with advanced NSCLC than in controls (p=0.024, p=0.028, respectively). The levels of survivin in NSCLC patients were comparable to controls. No correlation was found between HMGB1, survivin and VEGF concentrations and the histological type and staging of lung cancer. Similarly, no correlation was revealed between the concentrations of HMGB1, survivin and VEGF and the effect of chemotherapy. However, in patients with NSCLC, HMGB1 positevely correlated with survivin (R=0.814, p=0.007) before chemotherapy, and negatively with VEGF (R=-0.841, p=0.035) after chemotherapy. When the cut-off values of serum HMGB1, survivin and VEGF (2.38 ng/ml, 81.92 pg/ml, 443.26 pg/ml, respectively) were used, the prognoses of high and low groups were not different. Concluding, patients with NSCLC have a higher serum concentration of HMGB1 and VEGF, while survivin levels are comparable to healthy individuals. In our opinion, determination of HMGB1, survivin and VEGF concentrations has no clinical significance in the prognosis of the survival time in lung cancer.

Serum cathepsin K and cystatin C concentration in patients with advanced non-small-cell lung cancer during chemotherapy

A pathogenic implication of cathepsin K (Cath K) and its inhibitor - cystatin C (Cyst C) occur to be of growing importance in the mechanisms of tumor invasiveness in lung cancer. This study was conducted to investigate the prognostic role and the effects of chemotherapy on serum Cath K and Cyst C (ELISA) in patients with advanced stage non-small cell lung cancer (NSCLC). The study entered 40 patients (32 men) and 15 healthy volunteers (control group). Peripheral blood samples were taken before and after four cycles of chemotherapy. The mean serum Cyst C levels were significantly higher in patients with advanced NSCLC than in controls (p=0.003). The levels of Cath K in serum of NSCLC are comparable to those in controls. No correlation was found between Cath K and Cyst C concentrations and the histological type and staging of lung cancer. Patients with T4-stage had a lower level of Cyst C, than those with T2 (p=0.033). No correlation was found between the concentrations of Cath K, Cyst C and the effect of chemotherapy. However, Cyst C level positively correlated with serum creatinine concentration (R=0.535; p=0.005) in patients who responded to chemotherapy and with patients age (R=0.456; p=0.018) in whole group. When the cut-off values of serum Cath K and Cyst C (23.35 pmol/l, 1.29 mg/l, respectively) were used, the prognoses of high and low groups were not different. Concluding, patients with lung cancer have a higher serum concentration of Cyst C compared to healthy people. In our opinion, determination of Cath K and Cyst C concentrations has no clinical significance in the prognosis of the survival time in lung cancer.

Prognostic significance of DAPK and RASSF1A promoter hypermethylation in non-small cell lung cancer [NSCLC]

The epigenetic inactivation of tumor suppressor genes may play an important role in the development and progression of many cancer types, including lung cancer. Therefore, we investigated the association between the aberrant promoter methylation of 2 genes: the Death-Associated Protein Kinase (DAPK) and the Ras Association Domain Family 1A (RASSF1A) by using methylation-specific PCR, and the clinicopathological features and prognosis in 70 radically resected non-small cell lung cancers (NSCLCs). Hypermethylation of the DAPK and RASSF1A promoters was found in 24 (34%), and in 18 (26%) tumor DNA samples, respectively. Regarding different clinicopathological features of NSCLCs, the DAPK promoter methylation was more frequently observed in squamous cell carcinoma (46%) than in adenocarcinoma (25%) and large cell carcinoma (22%), but there were no significant statistical differences (p=0.3). On the other hand, a statistically significant trend was observed between the RASSF1A methylation and a histological type of tumor (p=0.06). 45% of adenocarcinoma tumors showed RASSF1A promoter methylation in comparison to 17% of squamous cell carcinomas and 22% of large cell carcinomas. When both markers were analyzed according to the tumor-node-metastasis (TNM) staging system, no statistically significant differences were observed between stage I, II and IIIa, and the DAPK (p=0.2) and RASSF1A methylation (p=0.1). In comparison, when stage I and II were grouped together and considered vs. stage IIIa, a significant association between RASSF1A methylation and the TNM was found (p=0.03). The group of patients with tumors showing DAPK promoter methylation had significantly poorer overall survival rates (p=0.02) than the patients with tumors that did not show DAPK promoter methylation. However, the association between the RASSF1A promoter methylation status and the overall survival rates was not statistically significant (p=0.48). In conclusion, this paper supports the importance of epigenetic gene regulation in lung cancer progression and prognosis.

Concentration of surfactant protein D, Clara cell protein CC-16 and IL-10 in bronchoalveolar lavage (BAL) in patients with sarcoidosis, hypersensivity pneumonitis and idiopathic pulmonary fibrosis.

The process of interstitial inflammation, often chronic, goes fluently from alveolitis through granuloma formation to irreversible fibrosis and lung remodeling. Eventually, the loss of functional alveolar units leads to chronic respiratory failure. The pneumoproteins (e.g. SP-D, CC-16) are considered to be markers of interstitial inflammation. We measured BAL concentration of SP-D, CC-16 and IL-10 in patients with sarcoidosis (27), IPF (7) and HP (9). The level of each marker was determined by ELISA specific kit. We found the highest SP-D and CC-16 BAL concentration in patients with the III stage of sarcoidosis (96,67 ng/ml and 31,78 ng/ml, respectively). The lowest SP-D concentration was observed in patients with IPF (76,49 ng/ml), and the lowest CC-16 concentration in patients with HP (21,39 ng/ml). The differences were not statistically significant. In the group of the III stage of sarcoidosis higher SP-D levels were related to higher BAL cytosis and higher percentage of BAL neutrophils, just the opposite as in the IPF and HP group. In the III stage of sarcoidosis and HP, the lower SP-D levels, the lower FEV1 and VC values. The results show, that in acute interstitial inflammation with larger parenchyma engagement (III stage of sarcoidosis) the levels of SP-D were higher then in chronic interstitial inflammation (IPF).

Lymphatic vessel invasion detected by the endothelial lymphatic marker D2-40 (podoplanin) is predictive of regional lymph node status and an independent prognostic factor in patients with resected esophageal cancer.

The discovery of markers to lymphatic endothelial cells and the development of novel antibodies to these markers have brought increasing attention to the lymphatics and progress in the understanding of lymphangiogenesis and cancer metastasis. In this study, we investigate the presence of lymphatic vessel invasion (LVI) detected by D2-40 immunohistochemical staining in resected esophageal cancer and correlated with clinicopathologic data and patient survival. Sixty nine patients, who had a primary resection of esophageal cancer, were analyzed by univariate and multivariate logistic regression, and univariate and multivariate survival analysis. The total rate of LVI was 72% (50/69). Positive LVI was significantly correlated with lymph node metastasis (p < 0.001), tumor size (p < 0.001), histological grading (p = 0.017), tumor depth (p = 0.001), and stage (p < 0.001). Multivariate logistic analysis identified LVI (p = 0.036) as a predictor of regional lymph node metastasis. On univariate survival analysis, patients with LVI had a significantly shorter disease-free survival, cancer-specific survival and overall survival. Multivariate analysis proved that LVI diagnosed by D2-40 is an independent prognostic factor of both disease-free survival (p = 0.04) and overall survival (p = 0.032) in resected esophageal cancer. These results show that LVI assessment identifies patients at high risk for regional lymph node metastasis and that LVI is an independent prognostic factor in patients with esophageal cancer.

Angiogenic Axis Angiopoietin-1 and Angiopoietin-2/Tie-2 in Non-Small Cell Lung Cancer: A Bronchoalveolar Lavage and Serum Study

Angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2), ligands for the Tie-2 receptor expressed on endothelial cells, play a critical role in angiogenesis, in concert with vascular endothelial growth factor (VEGF). Angiogenesis is important for tumor growth and development and also is implicated in the pathogenesis of interstitial lung diseases. The aim of this study was to evaluate the concentration of Ang-1, Ang-2, Tie-2, interleukin-18 (IL-18), transforming growth factor beta-1 (TGF β1), and VEGF domain in both serum and bronchoalveolar lavage fluid (BALF) of lung cancer patients before chemotherapy. We studied 45 non-small cell lung cancer (NSCLC) patients (M/F; 38/7; mean age 62 ± 4 years). The age-matched control groups consisted of 15 sarcoidosis (BBS), 15 hypersensivity pneumonitis (HP), and 15 healthy subjects. The patients with NSCLC had a significantly higher level of Ang-1 compared with the BBS and healthy subjects, and a higher level of Ang-2 compared with the healthy subjects in both serum and BALF. BALF level of IL-18 was lower in the NSCLC than that in the HP group, but higher than that in the BBS patients. Serum level of IL-18 was higher in the NSCLC than in the healthy subjects. The NSCLC group had lower VEGF in BALF than that in healthy subjects. Receiver-operating characteristics (ROC) curves were applied to find the cut-off the serum levels of Ang-1 and Ang-2 levels in BALF. We did not find any correlation between the levels of Ang-1, Ang-2, Tie-2, and the stage of tumor or treatment response (prospectively). We conclude that the angiogenic axis Ang-1 and Ang-2/Tie-2 may play an important role in lung cancer development and their concentrations may be a useful marker at the time of initial diagnosis of lung cancer.

Serum levels of VEGF-C, VEGF-D, and sVEGF-R2 in patients with lung cancer during chemotherapy.

The aim of this study was to assess serum levels of vascular endothelial growth factor C and D (VEGF-C, VEGF-D) and soluble VEGF receptor 2 (sVEGFR-2) in patients with lung cancer during chemotherapy. The study included 80 patients (64 men and 16 women; mean age 61.1) diagnosed histologically with lung cancer. Forty-four (55%) had non-small cell lung cancer (NSCLC) and 36 (45%) had small cell lung cancer (SCLC). Squamous cell carcinoma was established in 56% (25 patients) of all patients with NSCLC, adenocarcinoma in 20% (9 patients), and non-small cell lung cancer in 23% (10 patients). The control group consisted of 20 healthy volunteers. Peripheral blood samples were taken before and after four cycles of chemotherapy. VEGF-C, VEGF-D, and sVEGFR-2 levels were assessed by ELISA method. Serum levels of VEGF-C and VEGF-D were significantly higher in both NSCLC and SCLC groups in comparison with controls. VEGF-C concentration decreased after chemotherapy, whereas VEGF-D concentration was at the same level. No correlation was found between VEGF-C and VEGF-D concentrations and the effect of treatment. Patients with lung cancer and progression after chemotherapy (PD) had the higher concentration of sVEGFR-2 than patients with partial remission (PR). The levels of sVEGFR-2 were lower before and after treatment than in controls. No relation was found between VEGF-C, VEGF-D, and sVEGFR-2 concentrations and the histological type and staging of lung cancer. Summing up, serum concentrations of VEGF-C and VEGF-D were higher in patients with lung cancer both before and after chemotherapy than in healthy controls, whereas sVEGFR-2 concentration was lower than in healthy controls. An increase in concentration of sVEGFR-2 during chemotherapy may suggest progression of the disease. However, it requires further examination.

Morphological aspects of carcinogenesis in the lung.

The origins of the various histological types of primary lung cancer are not well understood. Numerous recent studies have indicated that lung cancer is not a result of a sudden transforming event in the bronchial and alveolar epithelium and in the neuroendocrine cells, but a multistep process in which a sequence of morphological and genetic changes is occurring. New modern technical approaches like fluorescence bronchoscopy techniques and microdissection, provide facilities to obtain valuable specimens for morphological and genetic verification of the sequentional changes in lung cancerogenesis. With their help, cells with morphologically recognized changes thought to be preneoplastic, may be removed and prepared for molecular and genetic studies. Therefore, the knowledge of the morphological aspects of lung preneoplastic lesions is crucial to make progress in molecular studies of lung carcinogenesis. Presently the knowledge about the sequence of molecular events in the lung carcinogenesis and their relationship to morphology is not perfect. In this review we will describe morphological aspects of various preneoplastic lesions occurring in the bronchial and bronchiolo-alveolar compartments including neuroendocrine cells.

Clinical Significance of HMGB-1 and TGF-β Level in Serum and BALF of Advanced Non-Small Cell Lung Cancer

Lung cancer is associated with poor prognosis. The aim of this study was to evaluate the clinical usefulness of HMGB-1 (high-mobility group protein B1) and TGF-β (transforming growth factor beta) in patients with advanced non-small cell lung cancer (NSCLC). We studied 45 patients with NSCLC prior to chemotherapy, 23 patients with Besnier-Boeck-Schaumann (BBS) disease (sarcoidosis), and 15 healthy volunteers. HMGB-1 and TGF-β levels were measured in serum and BALF samples using ELISA method. A higher serum HMGB-1 and TGF-β levels were in NSCLC patients compared with the other groups. TGF-β concentration in BALF was significantly higher in NSCLC than in healthy controls (p=0.047) but lower than in BBS (p=0.016). Serum HMGB-1 in NSCLC correlated with age and gender while its level in BALF was associated with distant metastasis. A higher levels of HMGB-1 in the serum of NSCLC patients with progressive disease was linked with shorter overall survival and disease-free survival. We found a positive correlation between HMGB-1 and TGF-β in BALF of IIIB NSCLC group and overall survival (p=0.04; p=0.003). Our findings confirmed that the measurement of HMGB-1 and TGF-β levels in serum and BALF of patients with NSCLC prior to treatment may have clinical usefulness and predict poor prognosis.

Serum Levels of Angiopoietin-1, Angiopoietin-2, and Their Receptor Tie-2 in Patients with Nonsmall Cell Lung Cancer During Chemotherapy

This pilot study was conducted to investigate the prognostic role and the effects of chemotherapy on serum angiogenic factors enzyme-linked immunosorbent assay consisting of Angiopoietin-1 and 2 (Ang-1, Ang-2) and their receptor Tie-2 in patients with advanced stage nonsmall cell lung cancer (NSCLC). Concentration of Ang-2 was higher in NSCLC (n= 40) than in healthy people (n= 15), whereas Ang-1 and Tie-2 were comparable. In our opinion determination of Ang-1, Ang-2, and Tie-2 concentrations have no clinical significance in the prognosis of the survival time in lung cancer and can not be used as a predictor of response to the chemotherapy.