Lymphatics-associated genes are downregulated at transcription level in non-small cell lung cancer.

The present study aimed to verify a possibility of ongoing lymphangiogenesis in non-small cell lung cancer (NSCLC) via examination of mRNA levels of a number of lymphangiogenesis-associated genes in tumors. It was hypothesized that transcriptional activation of these genes would occur in tumors that stimulate new lymphatic vessel formation. The study was performed on 140 pairs of fresh-frozen surgical specimens of cancer and unaffected lung tissues derived from NSCLC stage I-IIIA patients. mRNA levels were evaluated with the reverse transcription-quantitative polymerase chain reaction method and expressed as fold change differences between the tumor and normal tissues. Possible associations between expression and patient clinicopathological characteristics and survival were analyzed. In the NSCLC tissue samples, vascular endothelial growth factor (VEGF) C, VEGFD, VEGFR3, VEGFR2, VEGFR1, lymphatic vessel endothelial hyaluronan receptor 1, integrin subunit α 9, FOX2, neuropilin 2, fibroblast growth factor 2 genes were significantly downregulated (P<0.001 for all) compared with matched normal lung tissues, whereas mRNA levels for VEGFA, spleen associated tyrosine kinase, podoplanin, and prospero homeobox 1 genes were similar in both tissues. Neither lymph node status, nor disease pathological stage influenced expression, whereas more profound suppression of gene activities appeared to occur in squamous cell carcinomas compared with adenocarcinomas. The VEGFR1 mRNA expression level was significantly connected with patient survival in the univariate analysis, and was an independent prognostic factor for overall survival in the multivariate Cox's proportional hazards model (HR 2.103; 95% confidence interval: 1.005-4.401; P=0.049). The results support a hypothesis of absence of new lymphatic vessel formation inside growing NSCLC tumor mass, however do not exclude a possibility of lymphangiogenesis in narrow marginal tumor parts.

Serum soluble Fas ligand (sFasL) in patients with primary squamous cell carcinoma of the esophagus.

Esophageal carcinomas have been shown to express Fas ligand (FasL) and down-regulate Fas to escape from host immune surveillance. Circulating soluble FasL (sFasL) has been suggested to provide protection from Fas-mediated apoptosis. The aim of this study was to assess serum sFasL levels in esophageal cancer. The pretreatment levels of sFasL in the serum of 100 patients with esophageal squamous cell cancer and 41 healthy volunteers were determined by ELISA. Probability of survival was calculated according to the method of Kaplan-Meier. The prognostic influence of high and low level of sFasL was analyzed with the log-rank test. The mean serum level of sFasL in patients with esophageal cancer was significantly higher than that in healthy donors (1.567+/-1.786 vs 0.261+/-0.435, p<0.0001). The levels of serum sFasL were significantly higher in advanced stages (II vs IV p<0.034; III vs IV p<0.041; except II vs III p=0.281), patients with lymph node (N0 vs N1 p<0.0389) or distant (M0 vs. M1 p<0.0388) metastases and significantly lower in patients with well differentiated tumors (G1 vs G2 p<0.0272). The serum levels of soluble FasL were not related to gender, age, tumor size, T-stage, tobacco smoking and history of chronic alcohol intake. The survival difference between pretreatment high and low level of sFasL in surgery and chemio- and/or radiotherapy group was not statistically significant (p=0.525; p=0.840). Our results indicate that elevated serum sFasL levels might be associated with a disease progression in patients with esophageal squamous cell carcinoma.

MMP-10, MMP-7, TIMP-1 and TIMP-2 mRNA expression in esophageal cancer.

INTRODUCTION: Tissue inhibitors of metalloproteinases (TIMP) and the matrix metalloproteinases (MMP) are involved in the spread of cancer. METHODS: We have evaluated the matrix metalloproteinases' (MMP-10, MMP-7) and their inhibitors' (tissue inhibitors of metalloproteinases - TIMP-1, TIMP-2) mRNA expression in 61 esophageal cancer samples from patients who had undergone surgery, by using real-time quantitative RT-PCR, and correlated the results with the patient clinicopathologic features. RESULTS: MMP-10, MMP-7, TIMP-1, TIMP-2 were overexpressed in 73%, 85%, 55% and 42% of esophageal cancer samples, respectively. The expression of MMP-10, TIMP-1, and TIMP-2 correlated with the tumor size. The MMP-7 overexpression was associated with the tumour stage (I, II vs III, p=0.05) and lymph node metastasis (N0 vs N1, p=0.037). CONCLUSIONS: We conclude that in the resected esophageal cancer an increased mRNA expression of MMP-7, MMP-10 and TIMP-1 correlated with clinicopathologic features. We suggest that these genes may play a role during progression of the disease.

Prognostic value of vascular endothelial growth factor-C and podoplanin mRNA expression in esophageal cancer.

Vascular endothelial growth factor-C (VEGF-C), VEGF-D, VEGF receptor-3 (VEGFR-3) and podoplanin (PDPN) are involved in the spread of cancer. The current study evaluated VEGF-C, VEGF-D, VEGFR-3 and PDPN mRNA expression levels in 84 esophageal cancer samples from patients who had undergone surgery according to reverse transcription-quantitative polymerase chain reaction, and correlated the results with the clinicopathological features. The effects on lymph node metastasis and survival were identified by performing univariate and multivariate analyses. VEGF-C, PDPN, VEGF-D and VEGFR-3 were overexpressed in 52.4, 52.4, 32.1 and 51.2% of esophageal cancer samples, respectively. Furthermore, the expression of VEGF-C and PDPN was significantly correlated with lymph node metastasis, depth of tumor invasion and tumor stage (P<0.05). Logistic regression analysis identified tumor size (P=0.001), depth of invasion (P=0.002) and PDPN mRNA expression (P=0.022) as significant multivariable predictors of regional lymph node metastasis. Upon univariate survival analysis, the depth of tumor invasion, lymph node metastasis, histological grade, tumor stage, tumor size, residual tumor, and VEGF-C and PDPN mRNA expression were identified to be significant independent prognostic factors for overall survival (OS) time. Additionally, multivariate analysis identified tumor size (P=0.049), residual tumor (P<0.001) and PDPN mRNA expression (P=0.02) as independent factors for poor OS time. Thus, it was concluded that PDPN mRNA expression may serve as predictor for regional lymph node metastasis, and that VEGF-C and PDPN may be prognostic factors in patients with resected esophageal cancer.