ABSTRACT
<p><b>BACKGROUND</b>SYBR Green I is a non-probe real-time PCR method. It is quite convenient and its specificity is good. The aim of this study is to establish a quantitative SYBR Green I real-time PCR method for detection of PEDF gene in non-small cell lung cancer (NSCLC), and to investigate the relationship between PEDF mRNA expression and the clinicopathological characteristics.</p><p><b>METHODS</b>The target segments of PEDF and GAPDH proliferated by RT-PCR were diluted and used as the standard templates. PEDF mRNA was detected in 21 NSCLC tissues and matched paracancerous pulmonary tissues by relative quantitative method.</p><p><b>RESULTS</b>All the lung cancer tissues and the matched paracancerous pulmonary tissues had expression of PEDF mRNA. The relative level of PEDF mRNA expression was 0.5505±0.3590 (0.11-1.11) in the lung cancer tissues and 0.7219±0.2582 (0.29-1.31) in the matched paracancerous pulmonary tissues respectively (P=0.024). PEDF expression was significantly related to TNM stages (I-II vs III-IV, P=0.010) and the tumor size (T1 vs T2-4, P=0.007).</p><p><b>CONCLUSIONS</b>The established SYBR Green I quantitative real-time PCR method can successfully detect the expression of PEDF mRNA. The primary results show that PEDF may be a protective factor in oncogenesis and development of NSCLC.</p>
ABSTRACT
Tumour angiogenesis is a critical step in the growth, metastatic spread, and re-growth of many cancers. Tumour endothelial marker 8 [TEM-8] is a newly discovered molecule belongs to a family of endothelial markers that are raised during tumour angiogenesis. This study sought to examine the level of TEM-8 expression at the protein and mRNA level in human breast cancer tissues, and in a panel of human breast cancer cell lines, and also to determine if TEM-8 can be used as a suitable marker for identifying tumour associated micro-vessels. At the mRNA level more tumours showed positive TEM-8 expression compared to normal background tissues. TEM-8 was detected in a variety of breast cancer cell lines, endothelial cells [HEC V] and in a human fibroblast cell line [MRC5] at both the mRNA and protein level. Using immunohistochemistry the distribution of TEM-8 staining was more widespread in invasive breast cancer tissues compared to normal background tissues. Furthermore, the TEM-8 marker was found to be more discriminatory in identifying micro-vessels in tumour endothelium [2.8 +/- 0.83 vs. normal 1.66 +/- 0.52; P < 0.011], compared to the vWFA marker [1.61 +/- 0.54 vs. normal 2.71 +/- 0.76; P < 0.009]. Raised levels of TEM-8 were associated with shorter survival outcome, but were not correlated to disease free survival as shown by Kaplan-Meier and Cox regression analysis. We conclude that TEM-8 is a useful marker for identifying tumour associated micro-vessels and that elevated levels are associated with disease progression, which may have some bearing on the prognostic outcome in breast cancer. [Abbreviations ATR- antrax toxin receptor, cDNA -complementary dexoyribonucleic acid; EF-edema factor; LF- lethal factor; LeTx-lethal toxin; mRNA-messenger ribonucleic acid; PCR- polymerase chain reaction; Q-RT-PCR- quantitative real time polymerase chain reaction; RNA-ribonucleic acid; TEM-8 tumour endothelial marker-8]