Endothelial cell-anchored tissue factor pathway inhibitor regulates tumor metastasis to the lung in mice.

Tissue factor pathway inhibitor (TFPI) is a physiological inhibitor of the tissue factor (TF)-initiated coagulation pathway. Both circulating and tumor cell-associated TFPI significantly reduce tumor cell-induced coagulation activation and lung metastasis. However, the significance of endothelial cell-anchored TFPI in cancer biology remains largely unexplored. We generated mice with full-length disruption of TFPI (including TFPIα and TFPIß isoforms) in endothelial cells, using a Cre-LoxP system and gene inactivation (GI) strategy. Experimental pulmonary tumor metastasis models were used with TFPI-deficient mice to evaluate the role of endothelial cell-anchored TFPI in cancer progression. Finally, lung microvascular permeability and microenvironment were investigated. TFPI-deficient mice were viable and fertile, and showed decreased plasma TFPI levels and lung TFPI levels as compared with their control littermates. TFPI deficiency in endothelial cells promoted pulmonary tumor metastasis with an increased vascular permeability and altered lung microenvironment. Our observations suggest that endothelial cell-anchored TFPI controls lung tumor metastasis, and does so largely through the inhibition of local TF-induced thrombin generation and the regulation of the lung microenvironment in mice.

Grhl1 deficiency affects inner ear development in zebrafish.

Many genes that have been found to contribute to deafness are currently being studied. Some 87 non-syndromic hereditary deafness genes have been confirmed. Proteins associated with cochlear development have also been confirmed. Some of these proteins have important relationships with gap junctions (GJ) and tight junctions (TJ). However, the desmosome junction has received little attention due to controversy over whether it could be detected in the inner ear. GRHL1 is a conserved transcriptional regulator, and it is key to vertebrate desmosome formation. GRHL2 has been confirmed as a deafness gene at the DFNA28 locus. These two homologous proteins have similar sequences and functions. Here, a grhl1 down-regulated zebrafish model exhibited inner ear developmental malformations, including missing otoliths, disordered and abnormal numbers of hair cells in the inner ear and lateral line, and sound insensitivity. The mutant zebrafish swam in circles. Hair cell apoptosis was evident. Under electron microscopy, desmosomes in the otic sensory epithelium were found to be damaged. These defects were partially rescued by treatment with either GRHL1 or its target gene, DSG1. Collectively, these data are the first to indicate that grhl1 is important to the developing inner ear epithelia in zebrafish and that it acts via desmosome junction regulation.

Tissue factor pathway inhibitor-2 is critical in zebrafish cardiogenesis.

Human tissue factor pathway inhibitor-2 (Tfpi-2) is an extracellular matrix-associated Kunitz-type serine proteinase inhibitor and plays an important role in various cellular processes. We have previously shown that zebrafish Tfpi-2 (zTfpi-2) mainly expressed in the brain and heart of zebrafish, and it is involved in the development of central nervous system. Here, we identified zTfpi-2 as an evolutionarily conserved protein essential for zebrafish heart development, as embryos depleted of zTfpi-2 failed to undergo cardiogenesis. Changes of cardiogenic markers, vmhc, amhc and bmp4, confirmed zTfpi-2 knockdown caused cardiac defects, including retrenched ventricle, enlarged atrium and malformation of atrioventricular boundary. The sarcomeric organization was also disrupted by embryonic depletion of zTfpi-2, thus establishing the functional role of zTfpi-2 in cardiac contractility. In addition, hematopoietic defects were detected in the zTfpi-2-deficiency embryos. Importantly, injection of ztfpi-2 mRNA attenuated those cardiac and hematopoietic defects. Taken together, this study demonstrated a critical role of zTfpi-2 during embryonic cardiac development, as well as an important regulator of hematopoiesis.

[EGFR gene copy number, ERCC1 and BRCA1 protein expression and their relationship in non-small cell lung cancer].

OBJECTIVE: To evaluate the expression of epidermal growth factor receptor (EGFR) gene copy number and the expression of ERCC1 and BRCA1 proteins in patients with non-small-cell lung cancer (NSCLC) and the correlation between them. METHODS: The status of EGFR gene copy number was determined by in situ hybridization (FISH), and the expression of ERCC1 and BRCC1 proteins was examined by immunohistochemistry (IHC). The relationship of EGFR gene copy number with the expression of ERCC1 and BRCA1 and the clinical pathologic features were analyzed. RESULTS: FISH-positive EGFR expression was identified in 40 of 166 samples (24.1%). More FISH-positive EGFR in the female than male patients (31.9% vs. 18.6%, P = 0.048), and non-smoker than smoker (32.8% vs. 16.7%, P = 0.045). FISH-positive EGFR was not associated with age, pathological type, clinical stage and metestasis status (P > 0.05). The expression of ERCC1 protein was identified in 60 of 132 samples (45.5%). The expression of ERCC1 protein varied significantly in tumors of different pathological types (P = 0.046), but not associated with age, gender, clinical stage, metestatic status and smoking status (P > 0.05). The expression of BRCA1 protein was identified in 46 of 131 samples (35.1%). The expression of BRCA1 was not associated with age gender, pathological type, clinical stage, metestatic ststus and smoking status (P > 0.05). There was a moderate correlation between the expressions of ERCC1 and BRCA1 (r = 0.449, P < 0.001), but EGFR gene copy number was not correlated with the expression of ERCC1 or BRCA1 protein. CONCLUSIONS: FISH-positive EGFR expression is associated with gender and smoking status, but not correlated with the expression of ERCC1 and BRCA1 proteins. There is a moderate correlation between the expressions of ERCC1 and BRCA1.

[Correlation between pre-treatment anemia and prognosis in non-small cell lung cancer patients].

BACKGROUND AND OBJECTIVE: The patients with non-small cell lung cancer (NSCLC) might contract anemia, however, whether anemia is one of the independent prognostic factors to the patients with NSCLC is still controversial. So the aim of this study is to investigate the correlation between anemia and overall survival (OS) in patients with NSCLC. METHODS: 1 018 patients with operable NSCLC were retrospectively analyzed in our hospital from January 2000 to December 2008. RESULTS: The occurrence of anemia before operation was 252/1 018 (24.1%). The OS in NSCLC patients without anemia was (2 425.98 +/- 50.03) days, and the OS in patients with anemia was (2 107.15 +/-93.86) days. There was significant difference in the OS between them (P = 0.001). The patients with anemia in stage I had shorter survival time than those without anemia (P < 0.001). But there was no difference in other stage patients. TNM stage, gender, tumor size and lymph nodes metastasis were correlated with OS using Cox regression analysis. CONCLUSIONS: Anemia is correlated with survival in operable NSCLC patients. Moreover, it is an independent prognostic factor in NSCLC patients with stage I.

Establishment and characterization of a new drug surviving cell line Am1010, derived directly from muscle metastases of a human lung adenocarcinoma patient with multi-drug-resistance to cisplatin, taxol, and gefitinib.

AIM: To Characterize a new human lung cancer cell line Am1010, derived from drug-surviving cells (DSCs). METHODS: The Am1010 cell line was established after 4 cycles of chemotherapy from an arm muscle metastatic tumor of a patient diagnosed with lung adenocarcinoma. The cell line has been remained in continuous culture for more than one year during this study. RESULTS: The Am1010 cell line demonstrated in vitro multi-drug-resistance to cisplatin, taxol, and gefitinib. The Am1010 cell doubling time without drug treatment was 42.395 h. The IC(50) value of cisplatin was 4.299 micromol/L and >10 micromol/L for the Am1010 and P0318 (a cell line derived from non-DSCs) cells, respectively. The IC(50) value of taxol was 0.067 micromol/L and >1 micromol/L for the Am1010 and P0318 cells, respectively. The IC(50) value of gefitinib was 15.233 micromol/L and >70 micromol/L for Am1010 and P0318 cells, respectively. 11 genes involved in the focal adhesion and cell adhesion pathways were found to be differentially expressed. The cells of Am1010 have a significantly larger chromosome number than most lung cancer cell lines. CONCLUSION: This novel DSCs derived lung cancer cell line will be a valuable in vitro tool for the investigation of lung cancer drug resistance and metastasis.

Establishment of a lung cancer biobank of a southern chinese population.

There is an increasing need for establishment of biobanks for human cancer, which may facilitate basic and clinical researches as well as the development of novel approaches to early diagnostics, prevention and treatment, including personalized medicine. Herein we report the establishment of a lung cancer biobanks using biological samples from a lung cancer patient population in Southern China. Since 2007, we have collected lung cancer tissue from 1,054 patients with lung cancer, from whom 11,895 frozen tumor and normal matched tissues, 4,899 tissue paraffin blocks, and 3,562 blood serum samples were accumulated. The information on clinical manifestations, laboratory tests, and follow-up was maintained by an independent information management system, including three data sets. The primary data set included frozen tissue specimens, formalin-fixed paraffin-embedded tissues, blood specimens and clinical and long-term follow-up data. The secondary data set included DNA, RNA and proteins extracted from corresponding tissue specimens. And the tertiary data set contained improved genome, RNA and proteomics correlation analysis with relevant clinical and long-term follow-up data. The lung cancer biobanks is accessible to academic research and public services. To our best knowledge, this biobanks represents the first lung cancer tissue reservoir in China and should facilitate the basic and clinical research of this disease and developing diagnostic markers and novel therapeutic modalities.