Purpose: To investigate the diagnostic performance of circulating tumor cells (CTC) in discrimination between primary lung cancer and nonmalignant diseases as well as in prediction of distant metastasis. Patients and Methods: We prospectively evaluated CTCs in 7.5-mL samples of peripheral blood sampled from patients with a suspicion or a diagnosis of primary lung cancer. A semiautomated system was used to capture CTCs with an antibody against epithelial cell adhesion molecule. Results: Of 150 eligible patients, 25 were finally diagnosed as having nonmalignant disease, and 125 were diagnosed as having primary lung cancer with (n = 31) or without (n = 94) distant metastasis. CTCs were detected in 30.6% of lung cancer patients and in 12.0% of nonmalignant patients. CTC count was significantly higher in lung cancer patients than in nonmalignant patients, but a receiver operating characteristic (ROC) curve analysis showed an insufficient capability of the CTC test in discrimination between lung cancer and nonmalignant diseases with an area under ROC curve of 0.598 (95% confidence interval, 0.488-0.708; P = 0.122). Among lung cancer patients, CTC count significantly increased along with tumor progression, especially with development of distant metastasis. The area under ROC curve for CTC count in prediction of distant metastasis was 0.783 (95% confidence interval, 0.679-0.886; P < 0.001). When patients with one or more CTCs were judged as having metastatic disease, sensitivity and specificity of the CTC test were 71.0% and 83.0%, respectively. Conclusions: CTC is a useful surrogate marker of distant metastasis in primary lung cancer. (Clin Cancer Res 2009;15(22):6980-6)

In the present study, we analyzed genomic alterations of BRCA1-associated protein 1 (BAP1) in 23 malignant mesotheliomas (MMs), 16 epithelioid and seven non-epithelioid, consisting of 18 clinical specimens and five established cell lines. In examining these samples for homozygous deletions and sequence-level mutations, we found biallelic BAP1 gene alterations in 14 of 23 MMs (61%). Seven of these 14 MMs had homozygous deletions of the partial or entire BAP1 gene, another five had sequencelevel mutations, including small deletions, a nonsense mutation, and missense mutations with additional monoallelic deletions, and the remaining two had homozygous mutations without allelic loss. All but one of the 14 BAP1 gene mutations were found in the epithelioid-type MMs; BAP1 mutations were found in 13 of 16 epithelioid-type MMs, but in only one of seven non-epithelioid-type MMs (13/16 vs 1/7; P = 0.005). There was no BAP1 mRNA expression in MMs with biallelic deletion and repressed expression was confirmed in MM specimens with deletion/mutation as compared with Met5a, SV40-transformed normal mesothelial cells. Western blot showed that seven of eight epithelioid MMs analyzed were BAP1 negative. Immunostaining with anti-BAP1 antibody in normal lung tissues revealed clear nuclear staining of normal mesothelial cells. No nuclear staining was observed among BAP1 mutation-positive MM tumors, whereas nuclear staining was observed among BAP1 mutation-negative MM tumors. These results suggest that the lack of the tumor suppressor BAP1 may be more specifically involved in the pathogenesis of epithelioid MM rather than non-epithelioid MM, and would be useful for diagnosis of epithelioid-type MM. (Cancer Sci 2012; 103: 868-874) M alignant mesothelioma (MM) is an asbestos-related malignancy that arises primarily from surface serosal cells of pleural, peritoneal, and pericardial cavities. Although the use of asbestos has decreased in Western countries and Japan, the incidence of MM is expected to increase over the next few decades because of the long latency period (20-40 years) of this malignancy.(1) Although the prognosis of MM is generally poor, epithelioid-type MM has been reported to be associated with better prognosis than non-epithelioid types of MM.(2) Multiple modality approaches involving surgery with radiation, chemotherapy, or immunotherapy have generated favorable outcomes, particularly for patients with epithelioid-type MM.(3)

We documented a significant increase in CTC count in drainage PV blood after surgical manipulation, especially in tumours with lymphatic invasion. We are awaiting survival data at 5 year follow-up examination, which may provide clinical significance of the pvCTC-count.

No abstract

To develop a tool to obtain a high level of gene expression specifically in endothelial cells (ECs), we assessed enhancer activity of fragments in the first intron of the VE-cadherin gene using 3 different experimental systems: luciferase assay in the F2 EC line, green fluorescent protein (GFP) expression in ECs generated in embryonic stem (ES) cell differentiation culture, and GFP expression in transgenic mice. Although the 2.5-kbp (kilobase pair) 5 flanking sequence of the VE-cadherin gene is EC specific, adding 4 kbp of the 5 half of the first intron affected an enhancement of the gene expression level in all 3 assay systems. No other fragments tested in this study could confer such effects. Compared with other gene expression units, the unit described in this study would be the most optimum one available to date for EC-specific gene expression. Because this unit can express genes in VE-cadherin ؉ progenitors of hematopoietic cells but not in fully committed hematopoietic cells, it will be useful to manipulate specifically the uncommitted progenitor stage during hematopoietic cell differentiation. IntroductionEmbryonic stem (ES) cells are now established as a useful source of normal differentiated cells of various lineages, required not only for cell therapies of various degenerative diseases but also for preparing a panel of normal cells for drug screening. Although the advantage of this system lies in its ability to provide essentially unlimited amounts of cells, difficulties exist in steering the differentiation process to produce pure cell populations in culture. To overcome this problem, methods to purify the cells of interest from other unwanted populations have been developed. One such method uses cell sorting technology. We 1,2 and others 3 demonstrated that a set of surface markers can be used for defining and purifying various cell populations in the ES cell culture. Moreover, beta-galactosidase and green fluorescent protein (GFP) genes driven by specific promoters are also useful for purifying a subset of cells, particularly when applied in combination with surface markers. Another method is the drug-mediated selection of a particular lineage by which drug-resistant genes, driven by cell lineage-specific promoter units, are introduced into ES cells, thereby allowing the depletion of unwanted cell lineages during culture. 4 In both cases, the success of purification depends entirely on the availability of the lineage-specific promoter/enhancer unit. Moreover, such promoters are essential for expressing exogenous genes specifically in the cell lineage.In endothelial cell (EC) lineages, many promoters have been developed and shown useful for EC-specific gene expression. To our knowledge, however, most available promoters have a problem of expression in non-EC lineages. For instance, promoters of P-selectin and von Willebrand factor are active in megakaryocytes, 5,6 preproendothelin-1 is active in arterial smooth muscle cells, 7 and intercellular adhesion molecule-2 (ICAM-2) and platelet endothelial c...