The new adenocarcinoma classification has significant impact on death and recurrence in stage I lung adenocarcinoma. Patients with PL2 and micropapillary/solid predominant pattern have significant higher risk for recurrence. This information is important for patient stratification for aggressive adjuvant chemoradiation therapy.

Background: Metastasis is the most common cause of disease failure and mortality for non-small cell lung cancer (NSCLC) after surgical resection. Snail and TWIST1 are epithelial-mesenchymal transition (EMT) regulators which induce metastasis. Intratumoral hypoxia followed by stabilisation of hypoxia-inducible factor 1a (HIF-1a) promotes metastasis through regulation of certain EMT regulators. The aim of this study was to evaluate the prognostic value of HIF-1a, TWIST1 and Snail expression in patients with resectable NSCLC. Methods: A retrospective analysis of 87 patients with resectable NSCLC from Taipei Veterans General Hospital between 2003 and 2004 was performed using immunohistochemistry to analyse HIF-1a, TWIST1 and Snail expression. The association between HIF-1a, TWIST1 and Snail expression and patients' overall and recurrence-free survivals was investigated. Results: Overexpression of HIF-1a, TWIST1 or Snail was shown in 32.2%, 36.8% and 55.2% of primary tumours, respectively. Overexpression of HIF-1a, TWIST1 or Snail in primary NSCLCs was associated with a shorter overall survival (p = 0.005, p = 0.026, p = 0.009, respectively), and overexpression of HIF-1a was associated with a shorter recurrence-free survival (p = 0.016). We categorised the patients into four groups according to the positivity of HIF-1a/TWIST1/Snail to investigate the accumulated effects of these markers on survival. Coexpression of more than two markers was an independent prognostic indicator for both recurrence-free survival and overall survival (p = 0.004 and p,0.001, respectively, by multivariate Cox proportional hazards model). Conclusions: Co-expression of more than two markers from HIF-1a, TWIST1 and Snail is a significant prognostic predictor in patients with NSCLC.Non-small cell lung cancer (NSCLC) is the leading cause of cancer death worldwide. Surgical resection is the treatment of choice for early-stage NSCLC.

Dengue virus (DV) is a flavivirus and infects mammalian cells through mosquito vectors. This study investigates the roles of domain III of DV type 2 envelope protein (EIII) in DV bindingDengue virus (DV) is an arthropod-borne human pathogen that causes a serious public health threat in tropical and subtropical regions of the world (58). The World Health Organization reports that there are approximately 500,000 cases of dengue fever per year and that the infection rate is approximately 50 million per year (see reference 15 and literature cited therein). DV has four serotypes (DEN-1 to DEN-4) that cause diseases ranging from mild dengue fever to severe symptoms such as dengue hemorrhagic fever and dengue shock syndrome (17,25,27,70).The dengue viral genome is a single-stranded, positivestrand RNA with genome organization similar to those of other flaviviruses (47). DV infects a broad range of mammalian cell lines from several species in vitro but is transmitted to humans in vivo by mosquito vectors such as Aedes aegypti and Aedes albopictus (7). Primary human cells such as peripheral blood leukocytes, blood monocytes/macrophages, dendritic cells, and B lymphocytes could also be infected by DV (9,12,13,28,32,33,46,53,55,75,76,82).Previous studies indicate that cell surface heparan sulfates (HS) are involved in attachment of DV to mammalian cells including Vero, CHO, and human hepatoma cells (11,23,31,35,44,54). HS are repeating disaccharides composed of uronic acid or L-iduronic acid and a derivative of glucosamine that is variably O-sulfated (21). Extensive sulfate modification causes cell surface HS to be highly negatively charged. The biological roles of HS are quite diverse, including cell attachment and migration, compressive resilience of cartilage, control of fibrinogenesis, cell signaling, and virus infection (3). Many pathogenic microorganisms, including viruses, gram-positive and gram-negative bacteria, and parasites, attach to HS during entry into host cells (14,69,74). Since HS is ubiquitously expressed on many cell types and is commonly used by other pathogens to gain access into cells, an additional coreceptor has been postulated to explain the limited cell tropism of DV. This coreceptor may be related to a trypsin-sensitive protein or protein complex that was shown to play a role in virion binding to mammalian cells (18,52). Several candidate coreceptor proteins for DV have been suggested for many mammalian cell lines. These proteins are between 20 to 40 kDa and 60 to 90 kDa in size and bind to dengue virions in vitro (5,31,56,59,66). Recently, DC-SIGN, a dendritic cell surface lectin, was shown to mediate DV infection to primary dendritic cells (61,81). Nevertheless, the molecular mechanism by which DV enters these cells remains poorly characterized.Besides mammalian cells, mosquito cell lines that express cell surface proteins capable of binding to DV were also reported. In C6/36 cells, 40-to 45-kDa cell surface proteins were identified to bind 83). Additional candidate coreceptors on C6/36 cells w...

We previous reported that Sp1 recruits c-Jun to the promoter of the 12(S)-lipoxygenase gene in 12-myristate 13-acetate-treated cells. We now show that Sp1 that recruited HDAC1 to the Sp1/cJun complex was constitutively acetylated when cells were exposed to phorbol 12-myristate 13-acetate (PMA) (3 h). Prolonged stimulation of the cells with PMA (9 h), however, caused the dissociation of histone deacetylase 1 (HDAC1) and the deacetylation of Sp1, with the latter being able to recruit p300 that in turn caused the acetylation and dissociation of histone 3, thus enhancing the expression of 12(S)-lipoxygenase. We also overexpressed an Sp1 mutant (K703/A, lacking acetylation sites) in the cell and found that cells recruited more p300 and expressed more 12(S)-lipoxygenase. Taken together, our results indicated that Sp1 recruits HDAC1 together with c-Jun to the gene promoter, followed by deacetylation of Sp1 upon PMA treatment. p300 is then recruited to the gene promoter through the interaction with deacetylated Sp1 to acetylate histone 3, leading to the enhancement of the expression of 12(S)-lipoxygenase.

The role of specificity protein 1 (Sp1) in controlling gene expression in lung tumor development and metastasis is not well understood. In this study, we showed that the Sp1 level was highly increased and required for lung tumor growth in transgenic mice bearing Kras-induced lung tumors under the control of doxycycline. Furthermore, the Sp1 level was highly upregulated in lung adenocarcinoma cells with low invasiveness and in patients with stage I lung cancer. We also demonstrated that Sp1 was downregulated in lung adenocarcinoma cells with high invasiveness and in patients with stage IV lung adenocarcinoma. Moreover, Sp1 inversely regulated migration, invasion and metastasis of lung adenocarcinoma cells in vivo. In addition, a decrease in the Sp1 level in highly invasive lung adenocarcinoma cells resulted from instability of the Sp1 protein. Furthermore, overexpression of Sp1 in highly invasive lung adenocarcinoma cells increased expression of E-cadherin, a suppressor of metastasis, and attenuated the translocation of β-catenin into the cellular nucleus that leads to tumor malignancy. Taken together, Sp1 level accumulated strongly in early stage and then declined in late stage, which is important for lung cancer cell proliferation and metastasis during tumorigenesis.

In lung adenocarcinoma, the IASLC/ATS/ERS classification system has significant prognostic and predictive value regarding death and recurrence. Solid-predominant adenocarcinoma was also a significant predictor in patients undergoing adjuvant chemotherapy. Prognostic and predictive information is important for stratifying patients for aggressive adjuvant chemoradiotherapy.

The transcription factor Sp1 is ubiquitously expressed in different cells and thereby regulates the expression of genes involved in many cellular processes. This study reveals that Sp1 was phosphorylated during the mitotic stage in three epithelial tumor cell lines and one glioma cell line. By using different kinase inhibitors, we found that during mitosis in HeLa cells, the c-Jun NH 2 -terminal kinase (JNK) 1 was activated that was then required for the phosphorylation of Sp1. In addition, blockade of the Sp1 phosphorylation via inhibition JNK1 activity in mitosis resulted in the ubiquitination and degradation of Sp1. JNK1 phosphorylated Sp1 at Thr278/739. The Sp1 mutated at Thr278/739 was unstable during mitosis, possessing less transcriptional activity for the 12(S)-lipoxygenase expression and exhibiting a decreased cell growth rate compared with wild-type Sp1 in HeLa cells. In N-methyl-N-nitrosourea-induced mammary tumors, JNK1 activation provided a potential relevance with the accumulation of Sp1. Together, our results indicate that JNK1 activation is necessary to phosphorylate Sp1 and to shield Sp1 from the ubiquitin-dependent degradation pathway during mitosis in tumor cell lines. INTRODUCTIONThe transcription factor Sp1 is ubiquitously expressed in mammalian cells, and it is important in a variety of physiological processes, including cell cycle regulation, apoptosis, and differentiation (Firestone and Bjeldanes, 2003;Chu and Ferro, 2005;Wong et al., 2005;Deniaud et al., 2006). Sp1 binds specifically to the GC-rich promoter elements, via three C 2 H 2 -type zinc finger regions at the C terminus of Sp1, and it regulates the transcriptional activity of the target genes by using two major glutamine-rich transactivation domains localized, respectively, at the N terminus and the medial region (Suske, 1999;Bouwman and Philipsen, 2002). In addition, in Sp1, there are serine/threonine-rich sequences between the two transactivation domains that may be a target for posttranslational modification (Bouwman and Philipsen, 2002).The transcriptional activity of a transcription factor is determined at least by three factors: transactivational activity, DNA binding affinity, and protein level. Previous studies on the regulation of Sp1 activities focused mostly on transactivational activity, thereby allowing study of its interaction with other proteins and its DNA binding affinity. However, one of the apparent key elements regulating the activity of Sp1 is via its stability, which certainly needs to be explored and established. Recent studies revealed that the DNA binding ability, transactivational activity, and protein stability of Sp1 might be influenced by its posttranslational modifications such as sumoylation, glycosylation, ubiquitination, acetylation, and phosphorylation (Han and Kudlow, 1997;Mortensen et al., 1997;Wells et al., 2001;Ryu et al., 2003;Abdelrahim and Safe, 2005;Chu and Ferro, 2005;Hung et al., 2006;Spengler and Brattain, 2006). For example, Sp1 is sumoylated at Lys16, which might repress the transact...

The sigma-1 receptor (Sig-1R) chaperone at the endoplasmic reticulum (ER) plays important roles in cellular regulation. Here we found a new function of Sig-1R, in that it translocates from the ER to the nuclear envelope (NE) to recruit chromatin-remodeling molecules and regulate the gene transcription thereof. Sig-1Rs mainly reside at the ER-mitochondrion interface. However, on stimulation by agonists such as cocaine, Sig-1Rs translocate from ER to the NE, where Sig-1Rs bind NE protein emerin and recruit chromatin-remodeling molecules, including lamin A/C, barrier-to-autointegration factor (BAF), and histone deacetylase (HDAC), to form a complex with the gene repressor specific protein 3 (Sp3). Knockdown of Sig-1Rs attenuates the complex formation. Cocaine was found to suppress the gene expression of monoamine oxidase B (MAOB) in the brain of wild-type but not Sig-1R knockout mouse. A single dose of cocaine (20 mg/kg) in rats suppresses the level of MAOB at nuclear accumbens without affecting the level of dopamine transporter. Daily injections of cocaine in rats caused behavioral sensitization. Withdrawal from cocaine in cocaine-sensitized rats induced an apparent time-dependent rebound of the MAOB protein level to about 200% over control on day 14 after withdrawal. Treatment of cocaine-withdrawn rats with the MAOB inhibitor deprenyl completely alleviated the behavioral sensitization to cocaine. Our results demonstrate a role of Sig-1R in transcriptional regulation and suggest cocaine may work through this newly discovered genomic action to achieve its addictive action. Results also suggest the MAOB inhibitor deprenyl as a therapeutic agent to block certain actions of cocaine during withdrawal.T he endoplasmic reticulum (ER) plays important roles in cellular functions, including synthesis of proteins and regulation of Ca 2+ signaling between the ER and plasma membrane (1) and between the ER and mitochondria (2-4). However, because the ER interacts with other organelles in the cell, other functions related to the ER remain to be uncovered.The sigma-1 receptor (Sig-1R) (5-8) is an ER chaperone molecule that resides at the ER-mitochondrion interface referred to as the mitochondria-associated ER membrane (MAM), where the Sig-1R ensures proper ER-mitochondrion Ca 2+ signaling for cellular survival (3, 9), as well as sustains the activity of an ER stress sensor IRE1 at the MAM (10). The Sig-1R can translocate from the MAM to plasma membrane of the cell to regulate ion channels and receptors on the plasma membrane (8,(11)(12)(13)(14). Cocaine is a Sig-1R agonist (3) that causes the dissociation of Sig-1R from its cognate binding partner BiP (3,8), and consequently the translocation of Sig-1Rs to the plasma membrane, where Sig-1Rs interact with voltage-gated potassium channel subfamily A member 2 (Kv1.2) to shape the neuronal and behavioral responses to cocaine (15).Cocaine also causes the translocation of Sig-1Rs from the ER to the nucleus (16), where Sig-1Rs are shown to be present at the nuclear envelope (NE) (17). H...