bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhaled particles and lung cancer. Part A: Mechanisms
Authors: Knaapen Ad M. , Borm Paul J.A. , Albrecht Catrin , Schins Roel P.F.
Published: March 2004

Both occupational and environmental exposure to particles is associated with an increased risk of lung cancer. Particles are thought to impact on genotoxicity as well as on cell proliferation via their ability to generate oxidants such as reactive oxygen species (ROS) and reactive nitrogen species (RNS). For mechanistic purposes, one should discriminate between a) the oxidant‐generating properties of particles themselves (i.e., acellular), which are mostly determined by the physicochemical characteristics of the particle surface, and b) the ability of particles to stimulate cellular oxidant generation. Cellular ROS/RNS can be generated by various mechanisms, including particle‐related mitochondrial activation or NAD(P)H‐oxidase enzymes. In addition, since particles can induce an inflammatory response, a further subdivision needs to be made between primary (i.e., particle‐driven) and secondary (i.e., inflammation‐driven) formation of oxidants. Particles may also affect genotoxicity by their ability to carry surface‐adsorbed carcinogenic components into the lung. Each of these pathways can impact on genotoxicity and proliferation, as well as on feedback mechanisms involving DNA repair or apoptosis. Although abundant evidence suggests that ROS/RNS mediate particle‐induced genotoxicity and mutagenesis, little information is available towards the subsequent steps leading to neoplastic changes. Additionally, since most of the proposed molecular mechanisms underlying particle‐related carcinogenesis have been derived from in vitro studies, there is a need for future studies that evaluate the implication of these mechanisms for in vivo lung cancer development. In this respect, transgenic and gene knockout animal models may provide a useful tool. Such studies should also include further assessment of the relative contributions of primary (inflammation‐independent) and secondary (inflammation‐driven) pathways. © 2004 Wiley‐Liss, Inc.
Expression, regulation and function of autotaxin in thyroid carcinomas
Authors: Kehlen Astrid , Englert Nadine , Seifert Anja , Klonisch Thomas , Dralle Henning , Langner Jürgen , Hoang‐Vu Cuong
Published: March 2004

Autotaxin (ATX/NPP2) is a tumor cell motility‐stimulating factor that displays both a nucleotide pyrophosphatase/phosphodiesterase activity and a recently described lysophospholipase D (lysoPLD) activity. The precise function of ATX in tumor cells and the role of ATX in thyroid carcinoma remains unclear. We have quantified ATX mRNA expression in thyroid carcinoma cell lines and in tissues of patients with thyroid carcinomas. ATX gene activity was significantly higher in undifferentiated anaplastic thyroid carcinoma cell lines (UTC) and tumor tissues as compared to follicular thyroid carcinoma (FTC) cell lines, FTC tissues or goiter tissues that were used as a control. In the thyroid carcinoma cell line 1736, EGF and bFGF stimulated ATX mRNA expression, whereas the cytokines IL‐4, IL‐1β and TGF‐β reduced ATX transcriptional levels. FTC‐133 cells, stably transfected with an expression vector for ATX, showed a higher lysoPLD activity, a higher proliferation rate and an increased migratory behavior. In addition, ATX also displayed a paracrine stimulatory effect on the motility of different thyroid carcinoma cell lines. Overexpression of ATX in the stably transfected FTC‐133 resulted in down‐regulation of CD54/ intercellular adhesion molecule‐1 (ICAM‐1) gene expression and augmented gene activity of the pro‐angiogenic chemokine IL‐8. We conclude that ATX may be regarded as a new tissue marker for undifferentiated human thyroid carcinoma cells. ATX increases the proliferation and migration of thyroid carcinoma cell lines and may also affect the angiogenic potential of thyroid carcinoma cells. Further studies are needed to provide insight into the role of ATX in the normal and neoplastic thyroid gland. © 2004 Wiley‐Liss, Inc.
Effect of serum deprivation on constitutive production of granulocyte‐colony stimulating factor and granulocyte macrophage‐colony stimulating factor in lung cancer cells
Authors: Uemura Yoshiki , Kobayashi Makoto , Nakata Hideshi , Harada Ryoji , Kubota Tetsuya , Taguchi Hirokuni
Published: March 2004

We previously established 2 lung cancer cell lines, OKa‐C‐1 and MI‐4, which constitutively produce an abundant dose of granulocyte‐colony stimulating factor (G‐CSF) and granulocyte macrophage‐colony stimulating factor (GM‐CSF). Many other cases with G‐CSF or GM‐CSF producing tumors have been reported up to the present. However, the biological properties of the overproduction of G‐CSF and GM‐CSF by tumor cells have not been well known. Several reports demonstrated the presence of an autocrine growth loop for G‐CSF and GM‐CSF in nonhematopoietic tumor cells. We showed that exogenous G‐CSF and GM‐CSF stimulated cell growth in a dose‐dependent manner in OKa‐C‐1 and MI‐4 cells. We could detect the presence of G‐CSF and GM‐CSF receptors in both cell lines by RT‐PCR analysis. We have previously shown that inflammatory cytokines, tumor necrosis factor (TNF)‐α and interleukin (IL)‐1β enhance the expression of G‐CSF and GM‐CSF in the cell lines. However, the factors that regulate constitutive production of G‐CSF or GM‐CSF by tumor cells are still unknown well. In our study, we first reported that serum deprivation stimulated constitutive production of G‐CSF and GM‐CSF by lung tumor cells through activation of nuclear factor (NF)‐κB and p44/42 mitogen‐activated protein kinase (MAPK) pathway signaling. We suggest that G‐CSF and GM‐CSF constitutively produced by tumor cells could grow tumor itself and rescue tumor cells from the cytotoxicity of serum deprivation. © 2004 Wiley‐Liss, Inc.
Identifying progression‐associated genes in adult T‐cell leukemia/lymphoma by using oligonucleotide microarrays
Authors: Tsukasaki Kunihiro , Tanosaki Sakae , DeVos Sven , Hofmann Wolf K. , Wachsman William , Gombart Adrian F. , Krebs Johannes , Jauch Anna , Bartram Claus R. , Nagai Kazuhiro , Tomonaga Masao , Said Jonathan W. , Koeffler H. Phillip
Published: March 2004

Adult T‐cell leukemia/lymphoma (ATL) is associated with human T‐lymphotropic virus type‐1 (HTLV‐1). To understand the changes in expression that occur in the progression of chronic phase of ATL to acute crisis, the gene expression profiles of fresh ATL cells were compared in 4 pairs of samples (progression of chronic to acute phase in 3 patients, as well as 1 typical chronic phase sample vs. 1 typical acute phase sample) using high‐density oligonucleotide DNA arrays. We identified 203 genes that were commonly upregulated in acute vs. chronic phase samples including ribosomal proteins, proteosome subunits, eukaryotic translation factors, immunophilins, heat shock proteins and genes important for DNA replication. Additionally, we identified 91 commonly downregulated genes including immune molecules related to MHC and a phosphatase. Several of the genes were previously identified to be associated with the Tax protein of HTLV‐1. Some of the upregulated genes were located in amplified regions identified by comparative genomic hybridization in the corresponding chronic/acute ATL sample. Using real‐time quantitative PCR, we confirmed the array‐results in those specimens analyzed by microarray. These results demonstrated that distinct sets of genes that are known to be critical in cellular transformation and/or activation are up‐ or down‐regulated during the transition to the acute phase of ATL. © 2004 Wiley‐Liss, Inc.
Beta‐glucan enhanced killing of renal cell carcinoma micrometastases by monoclonal antibody G250 directed complement activation
Authors: Sier Cornelis F.M. , Gelderman Kyra A. , Prins Frans A. , Gorter Arko
Published: March 2004

Metastases from renal cell carcinomas (RCC) are resistant to radiation and chemotherapy but are relatively immunogenic. We have investigated the possibility to eliminate human RCC micrometastases using MAb G250. G250 penetrates human micrometastases completely in a spheroid model and induces complement deposition rapidly on the outmost cell layers. However, complement dependent cytotoxicity (CDC) was barely detected using either51chromium release assays or confocal microscopy, due to relatively low expression of the G250 antigen and the effect of membrane bound complement regulatory proteins. Addition of blocking anti‐CD59 MAbs enhanced formation of C5b‐9 and consequently complement mediated lysis (13%). Complement assisted cellular cytotoxicity (CACC) was not detectable, although the iC3b ligand and CR3 receptor were present on respectively target and effector cells. Addition of soluble β‐glucan induced the killing of MAb and iC3b opsonized spheroids by effector cells (6–21%). Despite a lower affinity for G250 antigen, a bispecific anti‐G250*anti‐CD55 MAb enhanced cell killing in spheroids comparable to the parental G250 MAb. Our results suggest that complement‐activating G250 in combination with anti‐mCRP MAbs is able to kill human RCC cells in micrometastasisin vitro. For CACC the presence of CR3‐priming β‐glucan seems to be obligatory.In vivo, bi‐MAb may be more effective as therapeutic agent due to its increased C5a generating properties. © 2004 Wiley‐Liss, Inc.
Inhibition of MUC1 expression by indole‐3‐carbinol
Authors: Lee Insong J. , Han Feng , Baek Jin , Hisatsune Akinori , Kim Kwang Chul
Published: March 2004

MUC1 is a large transmembrane glycoprotein overexpressed by a majority of carcinomas. High expression of MUC1 is associated with aggressive tumors, and MUC1 antigen is used as a marker to monitor disease progression in breast cancer patients. Several lines of evidence strongly suggest that the overexpression of MUC1 contributes to cancer progression and metastasis. In this report, we demonstrate that the naturally occurring cancer preventative, indole‐3‐carbinol (I3C), inhibits the expression of MUC1 in breast cancer cells. I3C inhibited both MUC1 mRNA and protein levels in a dose‐ and time‐dependent manner. This inhibition was seen in the estrogen responsive MCF‐7 cells as well as unresponsive MDA‐MB‐468 cells, indicating that the inhibitory pathway is independent of estrogen receptor. Gene expression studies using the human MUC1 gene promoter connected to a luciferase reporter demonstrated that I3C inhibits the transcription of the MUC1 gene. Promoter deletion studies indicate that the region containing up to 600 bp upstream (−600) of the initiation site is sufficient for inhibition by I3C. Furthermore, I3C represses the activation of transcription mediated by the region between −600 and −450 bp. A putative xenobiotic response element was located within this region but the binding of AhR/Arnt heterodimer to this site was undetectable by electrophoretic mobility shift assays. Our results may point to the existence of a novel pathway of transcriptional inhibition by I3C in cancer cells as well as a new mechanism of MUC1 gene inhibition. Our findings might have implications in the use of I3C as a preventative as well as a therapeutic agent for breast cancer. © 2004 Wiley‐Liss, Inc.
ABCG2 overexpression in colon cancer cells resistant to SN38 and in irinotecan‐treated metastases
Authors: Candeil Laurent , Gourdier Isabelle , Peyron Delphine , Vezzio Nadia , Copois Virginie , Bibeau Frederic , Orsetti Beatrice , Scheffer George L. , Ychou Marc , Khan Qasim A. , Pommier Yves , Pau Bernard , Martineau Pierre , Del Rio Maguy
Published: March 2004

Overcoming drug resistance has become an important issue in cancer chemotherapy. Among all known mechanisms that confer resistance, active efflux of chemotherapeutic agents by proteins from the ATP‐binding cassette family has been extensively reported. The aim of the present study was to determine the involvement of ABCG2 in resistance to SN38 (the active metabolite of irinotecan) in colorectal cancer. By progressive exposure to increasing concentrations of SN38, we isolated 2 resistant clones from the human colon carcinoma cell line HCT116. These clones were 6‐ and 53‐fold more resistant to SN38 than the HCT116‐derived sensitive clone. Topoisomerase I expression was unchanged in our resistant variants. The highest resistance level correlated with an ABCG2 amplification. This overexpression was associated with a marked decrease in the intracellular accumulation of SN38. The inhibition of ABCG2 function by Ko143 demonstrated that enhanced drug efflux from resistant cells was mediated by the activity of ABCG2 protein and confirmed that ABCG2 is directly involved in acquired resistance to SN38. Furthermore, we show, for the first time in clinical samples, that the ABCG2 mRNA content in hepatic metastases is higher after an irinotecan‐based chemotherapy than in irinotecan‐naive metastases. In conclusion, this study supports the potential involvement of ABCG2 in the development of irinotecan resistance in vivo. © 2004 Wiley‐Liss, Inc.
Flavaglines: A group of efficient growth inhibitors block cell cycle progression and induce apoptosis in colorectal cancer cells
Authors: Hausott Barbara , Greger Harald , Marian Brigitte
Published: March 2004

Flavaglines are flavonol–cinnamate‐derived cyclopenta[b]benzofurans, so far reported only for the genus Aglaia of the plant family Meliaceae. They represent a group of highly bioactive metabolites already known for their strong antileukemic activities. To assess their suitability as chemotherapeutic drugs in colorectal cancer, their cytostatic effects and the underlying mechanisms of action were analyzed in colorectal tumor cell lines. Aglaiastatin was the most active flavagline, inhibiting growth and inducing apoptosis at nanomolar concentrations in SW480 and HT29/HI1 carcinoma cells, while the premalignant adenoma cell lines VACO235 and LT97 as well as the normal intestinal epithelial cell line IEC18 were 1,000 times less sensitive (IC50 > 10 μM). In SW480 cells, aglaiastatin caused cell cycle block in early mitosis, demonstrated by a shift of cell cycle distribution 24 and 48 hr after addition of aglaiastatin and by an increased content of cyclin B after 6 hr together with a decreased level of cyclin A as early as 2 hr after exposure. In addition, induction of apoptosis could be shown by the characteristic morphology of apoptotic nuclei, loss of MMP and downmodulation of bclxl. Strong activation of p38 was observed after 2 hr of exposure, indicating that apoptosis may be induced via a p38‐mediated stress pathway. © 2004 Wiley‐Liss, Inc.
Synergistic effect of lymphotactin and interferon γ‐inducible protein‐10 transgene expression in T‐cell localization and adoptive T‐cell therapy of tumors
Authors: Huang Hui , Xiang Jim
Published: March 2004

The lack of efficient T‐cell infiltration of tumors is a major obstacle to successful adoptive T‐cell therapy. We have previously demonstrated that adenovirus (AdV)‐mediated transgene lymphotactin (Lptn) or IP‐10 expression in tumors can significantly enhance T‐cell tumor infiltration. In this study, active OVA‐specific CD8+ T cells were prepared by coculturing naive OVA‐specific CD8+ T cells from transgenic OT I mice with OVA‐I peptide‐pulsed dendritic cells in vitro. These XCR‐1‐ and CXCR3‐expressing T cells predominantly secreted IFN‐γ and displayed significant killing activity (84% at effector:target cell ratio of 1.5) against OVA‐expressing EG7 tumor cells through perforin‐mediated pathway. Our data also showed that chemokine Lptn and IP‐10 not only can chemoattract, but also stimulate proliferation of CD8+ T cells in vitro, and that a mixture of Lptn and IP‐10 can more efficiently chemoattract CD8+ T cells than either one of them. Furthermore, we demonstrated that the transferred CD8+ T cells detected in group of tumors treated with both AdVLptn and AdVIP‐10 (group a) are around 4 and 2 times more than that in groups of tumors treated with control AdVpLpA (group b) and either AdVIP‐10 (group c) or AdVLptn (group d), respectively. Around 87.5% of mice in group a were tumor‐free compared to the aggressive tumor growth in all 8 mice of group b and 25% or 37.5% cured mice seen in groups c and d (p < 0.05). Thus, our results indicate that enhancement of adoptive T‐cell therapy can be obtained by double tranmsgene Lptn and IP‐10 expression, which facilitates CD8+ T‐cell tumor localization through proliferation and chemoattraction of the transferred CD8+ T cells by in situ chemokine transgene expressions in the tumors. Collectively, our data provide solid evidence of a potent synergy between adoptive T‐cell therapy and adenovirus‐mediated Lptn and IP‐10 gene transfer into tumor tissues, which culminated in the T‐cell tumor localization and eradication of well‐established tumor masses. © 2004 Wiley‐Liss, Inc.
D‐PCa‐2: A novel transcript highly overexpressed in human prostate and prostate cancer
Authors: Weigle Bernd , Kiessling Andrea , Ebner Reinhard , Fuessel Susanne , Temme Achim , Meye Axel , Schmitz Marc , Rieger Michael A. , Ockert Detlef , Wirth Manfred P. , Rieber E. Peter
Published: March 2004

Identification of genes selectively expressed in tumors or individual tissues is a crucial prerequisite for molecular diagnosis and treatment of cancer by addressing molecular targets. By screening an expression database, we identified the novel gene D‐PCa‐2 (Dresden prostate carcinoma 2), which is highly overexpressed in normal prostate tissue and prostate carcinoma (PCa). The corresponding transcript contained an open reading frame of 453 nucleotides encoding a putative protein of 150 amino acids. A large part of exon 8 of the D‐PCa‐2 gene shows strong similarity to the high‐mobility‐group nucleosomal binding protein 2 (HMGN2) cDNA. The highly specific transcription of the D‐PCa‐2 gene in normal and malignant prostate tissues and in a few additional tumors was demonstrated by using multiple tissue dot blot, cancer profiling dot blot and real‐time PCR analyses. Examination of 18 pairs of tumorous and nontumorous prostate tissues from PCa patients by quantitative RT‐PCR revealed D‐PCa‐2 transcripts in all specimens. The potential usefulness of D‐PCa‐2 as a sensitive marker for metastatic prostate carcinoma cells in lymph nodes was demonstrated by the detection of one LNCaP cell in 1 × 105 normal lymph node cells using real‐time RT‐PCR. Examination of 22 lymph nodes from PCa patients either containing metastatic prostate cancer cells or diagnosed as cancer‐free was in full concordance with histopathologic diagnoses. These results validate D‐PCa‐2 as a transcript with high tissue specificity and with a potential application in the diagnosis of PCa. © 2004 Wiley‐Liss, Inc.
Invariant natural killer T cells are preserved in patients with glioma and exhibit antitumor lytic activity following dendritic cell‐mediated expansion
Authors: Dhodapkar Kavita M. , Cirignano Barbara , Chamian Francesca , Zagzag David , Miller Douglas C. , Finlay Jonathan L. , Steinman Ralph M.
Published: March 2004

Brain tumors carry a poor prognosis, and newer approaches to their therapy are urgently needed. Natural killer T (NKT) cells are distinct innate lymphocytes with antitumor potentials. Defects in NKT cell function have been observed in patients with other forms of cancer. Here we show that both the frequency and interferon‐γ‐producing function of NKT cells are well preserved in adult patients with glioma (n = 9) and comparable to findings in healthy controls (n = 9). These cells can be readily expanded in culture using autologous mature dendritic cells loaded with the NKT ligand, α‐galactosyl ceramide. The expanded NKT cells from glioma patients are functional and, importantly, kill glioma cells in a ligand‐ and CD1d‐dependent manner. Expression of CD1d is detected both on primary glioma cells as well as endothelial cells in infiltrating new blood vessels by immunohistochemistry of glioma tissue sections. These data suggest that targeting NKT cells may provide a novel strategy for immunotherapy of glioma. © 2004 Wiley‐Liss, Inc.
Phenotypic alterations induced by the Hong Kong‐prevalent Epstein‐Barr virus‐encoded LMP1 variant (2117‐LMP1) in nasopharyngeal epithelial cells
Authors: Lo Angela Kwok Fung , Huang Dolly P. , Lo Kwok Wai , Chui Yiu Loon , Li Hoi Ming , Pang Jesse Chung Sean , Tsao Sai‐Wah
Published: March 2004

Epstein‐Barr virus (EBV) is closely associated with nasopharyngeal carcinoma (NPC), a common cancer in Hong Kong. The EBV‐encoded LMP1 protein is believed to play an important role in cell transformation. We have previously identified a prevalent LMP1 variant (2117‐LMP1) that is expressed in 86% of primary NPC in Hong Kong. In this study, the biologic phenotypes induced by 2117‐LMP1 were compared with those of the prototypic B95.8‐LMP1 in an immortalized nasopharyngeal epithelial cell line, NP69. The 2117‐LMP1 could induce cell proliferation and resistance to apoptosis induced by growth factor deprivation. Expression of 2117‐LMP1 also suppressed expression of p16, p21 and Bax but induced expression of CDK2 and A20. Compared with B95.8‐LMP1, 2117‐LMP1 could induce a higher migration ability in NP69 cells but was less efficient in inducing morphologic changes, anchorage‐independent growth and cell invasion. Relatively weaker ability of 2117‐LMP1 than B95.8‐LMP1 in upregulation of vimentin, VEGF and MMP9 as well as in downregulation of E‐cadherin was observed. 2117‐LMP1 could activate higher level of NF‐κB activity in HEK 293 cells than B95.8‐LMP1. The present study supports a role of 2117‐LMP1 in NPC development by enhancing cell proliferation, cell death inhibition and migration in premalignant nasopharyngeal epithelial cells. Furthermore, our study reveals significant functional differences between 2117‐LMP1 and the prototypic B95.8‐LMP1. Our results provide insights into the pathologic significance of this prevalent LMP1 variant, 2117‐LMP1, in the development of NPC in the Hong Kong population. © 2004 Wiley‐Liss, Inc.
Expression of GLUT‐1 glucose transfer, cellular proliferation activity and grade of tumor correlate with [F‐18]‐fluorodeoxyglucose uptake by positron emission tomography in epithelial tumors of the ovary
Authors: Kurokawa Tetsuji , Yoshida Yoshio , Kawahara Kazumi , Tsuchida Tatsuro , Okazawa Hidehiko , Fujibayashi Yasuhisa , Yonekura Yoshiharu , Kotsuji Fumikazu
Published: March 2004

We evaluated whether tracer FDG uptake, quantified as an SUV by PET in ovarian epithelial tumors, correlates with clinical stage, tumor grade, cell proliferation and glucose metabolism, all of which are biomarkers for response to chemotherapy, prognosis and overall survival in ovarian cancer patients. Seventeen patients suspected of having ovarian cancer by physical examination, tumor marker analysis and anatomic imaging (such as sonography, CT and/or MRI) underwent whole‐body FDG‐PET within the 2 weeks prior to surgery. Seventeen epithelial ovarian tumor specimens (13 malignant tumors, 5 at stage I, 2 at stage II, 6 at stage III; 2 borderline tumors; and 2 benign lesions) were available for pathologic evaluation. They were graded histopathologically, and immunohistochemistry for MIB‐1 (proliferation index marker) and GLUT‐1 was performed. Correlation between FDG uptake and clinical stage, GLUT‐1 expression, MIB‐1 LI and histologic grading score was determined. No positive correlation was observed between FDG uptake and clinical stage (p = 0.14). Intensity of GLUT‐1 expression (r = 0.76, p = 0.001), MIB‐1 LI (r = 0.457, p = 0.014) and histologic grading score (r = 0.692, p = 0.005) showed statistically significant positive correlations with FDG uptake. Stepwise logistic regression analysis revealed that expression of GLUT‐1 transporters was the strongest parameter (r = 0.760, p = 0.0004) by which to predict positive FDG uptake. Therefore, glucose consumption, as determined by analysis of SUVs in FDG‐PET, may be a noninvasive biomarker for ovarian epithelial tumors. © 2004 Wiley‐Liss, Inc.
RASSF1A is a target tumor suppressor from 3p21.3 in nasopharyngeal carcinoma
Authors: Chow Lillian Shuk‐Nga , Lo Kwok‐Wai , Kwong Joseph , To Ka‐Fai , Tsang Kam‐Sze , Lam Ching‐Wan , Dammann Reinhard , Huang Dolly P.
Published: March 2004

Deletion on the short arm of chromosome 3 is one of the most important genetic abnormalities in the tumorigenesis of nasopharyngeal carcinoma (NPC). Both physical mapping and functional studies have targeted an NPC‐related tumor suppressor gene(s) to chromosome 3p21.3. We have reported recently that RASSF1A gene, located on a 120‐kb minimal deletion region on 3p21.3, was frequently inactivated by promoter hypermethylation in NPC. We further confirmed that RASSF1A is the critical target tumor suppressor from 3p21.3, with the evidence that loss of expression and aberrant methylation of the other 8 candidate genes/transcripts (HYAL2, FUS1, RASSF1C, BLU, NPRL2, 101F6, PL6 and CACNA2D2) in this 120‐kb region were rare in NPC samples. The contribution of RASSF1A in NPC tumorigenesis was investigated by restoring its expression in a RASSF1A deficient cell line, C666‐1. Transient transfection of wild‐type RASSF1A resulted in marked growth inhibition in NPC cells. Isolated stable clones expressing wild‐type RASSF1A demonstrated retarded cell proliferation in vitro. Soft‐agar assay also showed decreased number and sizes of colony formed in these clones. In vivo nude mice assay demonstrated the dramatic reduction of tumorigenic potential in the RASSF1A‐transfected clones. Our results provide strong evidence to support RASSF1A as a target tumor suppressor gene on 3p21.3 in NPC. © 2004 Wiley‐Liss, Inc.
Women smoking and testicular cancer: One epidemic causing another?
Authors: Pettersson Andreas , Kaijser Magnus , Richiardi Lorenzo , Askling Johan , Ekbom Anders , Akre Olof
Published: March 2004

In many countries the incidence of testicular cancer has increased epidemically, but the aetiology remains obscure. Maternal smoking during pregnancy has been suggested to be a cause, but a satisfactorily valid assessment of the hypothesis is still lacking. To evaluate the epidemiological relevance, we assessed the ecological correlation between female smoking habits and testicular cancer incidence in the Nordic countries. Data on smoking prevalence among women in 5‐year birth cohorts 1910–1940, at 25–29 years of age, were obtained through past studies of smoking habits in Sweden, Denmark, Norway and Finland. Testicular cancer incidence in the presumed offspring birth cohorts 1938–1968, was calculated using data from the respective cancer registries. For comparison, a similar analysis of male smoking and testicular cancer in the presumed offspring cohorts was carried out. The Pearson correlation coefficient between female smoking prevalence and testicular cancer incidence in the generation of the presumed offspring was 0.9 (p < 0.0001) for all countries combined. With the exception of Finland, the country‐specific correlations were of the same magnitude. In multivariate analysis, there was no correlation with male smoking. The strong geographical and temporal correlations between female smoking and testicular cancer indicate that if smoking during pregnancy is a cause of testicular cancer in the offspring, part of the increasing trends could be explained, and more importantly: avoided, by primary prevention. © 2004 Wiley‐Liss, Inc.
Role for dipeptidyl peptidase IV in tumor suppression of human non small cell lung carcinoma cells
Authors: Wesley Umadevi V. , Tiwari Shakuntala , Houghton Alan N.
Published: March 2004

Lung cancer is the leading cause of cancer death. Lung cancers produce a variety of mitogenic growth factors that stimulate tumor cell proliferation and migration. The cell surface protease, dipeptidyl peptidase IV (DPPIV), is involved in diverse biologic functions, including peptide‐mediated cellular growth and differentiation. DPPIV is expressed in various normal tissues, including lung tissue, and its expression is lost in many types of human cancers. DPPIV expression and its enzymatic activity are detected in normal bronchial and alveolar epithelium but different histologic subtypes of lung carcinomas lose DPPIV expression. To investigate the role of DPPIV in lung carcinoma, we examined the expression of DPPIV at both mRNA and protein levels in non small cell lung cancer (NSCLC) cell lines and normal human bronchial epithelial cells. DPPIV expression was detectable in normal lung epithelial cells, but was absent or markedly reduced in all NSCLC cell lines at both mRNA and protein levels. Restoration of DPPIV expression in NSCLC cells resulted in profound morphologic changes, inhibition of cell proliferation, anchorage‐independent growth, in vitro cell migration and tumorigenicity in nude mice. DPPIV reexpression also correlated with increased p21 expression, leading to induction of apoptosis and cell cycle arrest in G1 stage. These effects were accompanied by increased expression of cell surface proteins, fibroblast‐activating protein (Fapα) and CD44 that are associated with suppression of tumor growth and metastasis. Thus, DPPIV functions as a tumor suppressor, and its downregulation may contribute to the loss of growth control in NSCLC cells. © 2004 Wiley‐Liss, Inc.
Human mena protein, a serex‐defined antigen overexpressed in breast cancer eliciting both humoral and CD8+ T‐cell immune response
Authors: Di Modugno Francesca , Bronzi Giovanna , Scanlan Matthew J. , Del Bello Duilia , Cascioli Simona , Venturo Irene , Botti Claudio , Nicotra Maria Rita , Mottolese Marcella , Natali Pier Giorgio , Santoni Angela , Jager Elke , Nisticò Paola
Published: March 2004

Screening of a cDNA expression library from a primary breast tumor with the autologous patient serum led to the isolation of 6 cDNA clones corresponding to 3 different genes, including a novel gene that maps to chromosome 1 and encodes the human homologue of mouse Mena (hMena, cDNA clone RMNY‐BR‐55), a protein of the Ena/VASP family involved in the regulation of cell motility and adhesion. A cancer‐restricted antibody response against hMena was demonstrated, since 18/93 cancer patient sera, the majority (10/52) from breast cancer, showed anti‐hMena‐specific IgG, while no antibodies were present in healthy donors. When hMena protein expression was analyzed by Western blot and immunohistochemistry, the antigen was overexpressed in the majority of breast cancer cell lines and in 75% of primary breast tumor lesions evaluated. Furthermore, when HLA‐A2‐restricted peptides from the hMena sequence were used to stimulate CD8+ T cells, an hMena‐specific response was found in 9 out of 12 HLA‐A2+ breast cancer patients. In 4 patients, this cell‐mediated immune response was concomitant with antibody response to hMena. Furthermore, an hMena‐specific T‐cell line was established from an HLA‐A2+ breast cancer patient whose primary tumor lesion overexpressed the hMena protein. The present findings highlight the emerging role that overexpression of cytoskeleton regulatory components may have in the induction of a specific antitumor immune response. © 2004 Wiley‐Liss, Inc.
c‐jun NH2‐terminal kinase pathway is involved in constitutive matrix metalloproteinase‐1 expression in a hepatocellular carcinoma‐derived cell line
Authors: Sugioka Yoshihiko , Watanabe Tetsu , Inagaki Yutaka , Kushida Miwa , Niioka Maki , Endo Hitoshi , Higashiyama Reiichi , Okazaki Isao
Published: March 2004

Transcription factor c‐Jun serves for cellular proliferation, survival, differentiation and transformation and is recognized as an important factor in cancer development, including hepatocellular carcinoma (HCC). The purpose of present study is to determine the involvement of c‐Jun in matrix metalloproteinase‐1 (MMP‐1) expression, which is previously reported by us to be expressed only in the early stage of human HCC showing stromal invasion. Of 5 human HCC cell lines examined, only HLE cells revealed mRNA and protein expression as well as enzymatic activity of MMP‐1. Transient transfection of an MMP‐1 promoter/luciferase construct (including 4.4 kb full promoter region) into HLE and HCC‐T cells (MMP‐1 nonproducer) showed that high promoter activity was observed only in HLE cells without inducers, and that this promoter activity was still observed when a shorter 0.6 kb proximal promoter construct was transfected. The 0.6 kb promoter region contained 3 AP‐1 sites, and c‐jun mRNA was constitutively expressed in HLE cells without inducers. Furthermore, phosphorylated c‐Jun and c‐Jun NH2‐terminal kinase (JNK) were detected in HLE cells. Promoter activity of the 0.6 kb construct was suppressed with SP600125, a potent inhibitor of JNK, but not with PD98059 and SB203580, potent inhibitors of MEK1/2 and p38, respectively. The inhibitory effect of SP600125 was also observed at protein expression level and in enzymatic activity of MMP‐1. Taken together, this study suggests that the JNK pathway is involved in the expression of MMP‐1 in HCC cells and may represent a new functional role of c‐Jun for HCC development. © 2004 Wiley‐Liss, Inc.
First‐principles pair potentials across the metal–ceramic interface
Authors: Han Xiaoping , Zhang Yue , Xu Huibin
Published: March 2004

Systematic approaches are presented to invert the first‐principle binding energy to obtain the pair potentials across the metal/ceramic interfaces. The conformation characteristics of two virtual interface models suitable for the inversion are summarized: (1) the positions of one kind of ceramic ions stay unchanged, but those of the other change; or, (2) there is no change in the relative positions of the ceramic ions to the metal atoms but the positions of two kinds of ceramic ions are all exchanged each other. As an example, we derive the pair potentials of NiZr4+ and NiO2− across the Ni/ZrO2 interface with this method, which exhibit comparable accuracy to the first‐principle results. This provides direct evidence that the interfacial potentials obtained from this method can reasonably describe the general features of bondings across the metal–ceramic interface. The limitation of this method is also discussed. © 2004 Wiley Periodicals, Inc. J Comput Chem 25: 968–973, 2004
Ensemble variance in free energy calculations by thermodynamic integration: Theory, optimal “Alchemical” path, and practical solutions
Authors: Blondel Arnaud
Published: March 2004

Thermodynamic integration is a widely used method to calculate and analyze the effect of a chemical modification on the free energy of a chemical or biochemical process, for example, the impact of an amino acid substitution on protein association. Numerical fluctuations can introduce large uncertainties, limiting the domain of application of the method. The parametric energy function describing the chemical modification in the thermodynamic integration, the “Alchemical path,” determines the amplitudes of the fluctuations. In the present work, I propose a measure of the fluctuations in the thermodynamic integration and an approach to search for a parametric energy path minimizing that measure. The optimal path derived with this approach is very close to the theoretical minimum of the measure, but produces nonergodic sampling. Nevertheless, this path is used to guide the design of a practical and efficient path producing correct sampling. The convergence with this practical path is evaluated on test cases, and compares favorably with that of other methods such as power or polynomial path, soft‐core van der Waals, and some other approaches presented in the literature. © 2004 Wiley Periodicals, Inc. J Comput Chem 25: 985–993, 2004
On the applicability of the HSAB principle through the use of improved computational schemes for chemical hardness evaluation
Authors: Putz Mihai V. , Russo Nino , Sicilia Emilia
Published: March 2004

Finite difference schemes, named Compact Finite Difference Schemes with Spectral‐like Resolution, have been used for a less crude approximation of the analytical hardness definition as the second‐order derivative of the energy with respect to the electron number. The improved computational schemes, at different levels of theory, have been used to calculate global hardness values of some probe bases, traditionally classified as hard and soft on the basis of their chemical behavior, and to investigate the quantitative applicability of the HSAB principle. Exchange acid‐base reactions have been used to test the HSAB principle assuming the reaction energies as a measure of the stabilization of product adducts. © 2004 Wiley Periodicals, Inc. J Comput Chem 25: 994–1003, 2004
Conformational fluctuations and electronic properties in myoglobin
Authors: Aschi Massimiliano , Zazza Costantino , Spezia Riccardo , Bossa Cecilia , Di Nola Alfredo , Paci Maurizio , Amadei Andrea
Published: March 2004

In this article we use the recently developed perturbed matrix method (PMM) to investigate the effect of conformational fluctuations on the electronic properties of heme in Myoglobin. This widely studied biomolecule has been chosen as a benchmark for evaluating the accuracy of PMM in a large and complex system. Using a long, 80‐ns, molecular dynamics simulation and unperturbed Configuration Interaction (CISD) calculations in PMM, we reproduced the main spectroscopic features of deoxy‐Myoglobin. Moreover, in line with our previous results on a photosensitive protein, this study reveals a clear dynamical coupling between electronic properties and conformational fluctuations, suggesting that this correlation could be a general feature of proteins. © 2004 Wiley Periodicals, Inc. J Comput Chem 7: 974–984, 2004
Applied cognitive psychology: Implications of cognitive psychology for clinical psychology and psychotherapy
Authors: Eysenck Michael W.
Published: March 2004

Cognitive psychology has made numerous contributions to clinical psychology, and these contributions are considered especially with reference to the anxiety disorders. It is argued that there are four major contributions that can be identified. First, the cognitive approach has led to the development of complex models showing the main cognitive processes and structures of relevance to an understanding of anxiety disorders. Second, controlled laboratory studies permit a more detailed investigation of cognitive biases in anxious patients than generally is feasible in more naturalistic settings. Third, the cognitive approach provides relevant evidence with respect to the issue of whether cognitive biases play a role in the development and maintenance of anxiety disorders. Fourth, the enhanced understanding of the anxiety disorders that has arisen from the cognitive approach has had beneficial effects on therapeutic practice in a number of significant ways. In sum, it is claimed that clinical psychology has benefited considerably from cognitive theory and research. © 2004 Wiley Periodicals, Inc. J Clin Psychol.
Cognitive factors in health psychology and behavioral medicine
Authors: Montgomery Guy H.
Published: March 2004

Research in health psychology and behavioral medicine has increased exponentially over the past three decades. Prevention and control efforts across a variety of diseases have recognized that individual difference factors, belief systems, attitudes, behaviors, and environments can affect an individual's health. This work continues, as there has been no abatement in the rate at which aversive, chronic, and life‐threatening diseases continue to affect millions of Americans each year. The purpose of this review is to discuss the influence of cognitive factors on health psychology and behavioral medicine using the cancer literature as an example. The main discussion focuses on the impact of cognitive constructs on behavioral medicine and health psychology in cancer, which appears to be widespread. The article concludes with some implications for future research in the area. © 2004 Wiley Periodicals, Inc. J Clin Psychol.
An essential role for the His‐8 residue of the SDF‐1α–chimeric, tropism‐redirected Env protein of the Moloney murine leukemia virus in regulating postbinding fusion events
Authors: Katane Masumi , Fujita Rika , Takao Eiko , Kubo Yoshinao , Aoki Yasunobu , Amanuma Hiroshi
Published: March 2004

Background

To use retroviral vectors for the cell‐specific delivery of genes, it is necessary to redirect their receptor tropism to cell‐specific receptors. Previously, we reported that a Moloney murine leukemia virus (MLV) retroviral vector containing a human stromal‐derived factor‐1α (SDF‐1α)–chimeric envelope protein (Env) (S3) acquired the ability to transduce human cells via CXCR4, the cognate receptor for SDF‐1α, while retaining the ability to transduce mouse cells via mCAT1.

Methods

We constructed expression plasmids for derivatives of the S3 Env protein; S3‐D84K containing an Asp‐84‐to‐Lys (D84K) substitution, S3‐H8R‐D84K containing D84K and an additional His‐8‐to‐Arg substitution, and S3‐D84K‐RY containing D84K and additional Gln‐227‐to‐Arg plus Asp‐243‐to‐Tyr substitutions which have been suggested to suppress the loss of function of His‐8. Cellular expression, virion incorporation, and entry functions of these derivatives were investigated.

Results

All three derivatives were incorporated into virions. The S3‐D84K vector lost its ecotropism, but could transduce CXCR4‐expressing human and mouse cells at titers of 103 to 104 colony‐forming units (cfu)/ml. The S3‐H8R‐D84K vector did not show transduction, although its Env protein could bind to CXCR4. The transduction titer of the S3‐D84K‐RY vector via CXCR4 was slightly lower than that of the S3‐D84K vector. These results indicate that the His‐8 residue of the S3‐D84K Env protein is indispensable and may be fully functional in postbinding membrane fusion.

Conclusions

Insertion of a ligand at Pro‐79 of the Moloney MLV Env protein has proved to be a valuable strategy for constructing direct targeting retroviral vectors, since it permits the formation of a redirected Env protein without ecotropism, and it does not disrupt the function of the essential His‐8 residue. Copyright © 2004 John Wiley & Sons, Ltd.
LIM kinase‐2 targeting as a possible anti‐metastasis therapy
Authors: Suyama Eigo , Wadhwa Renu , Kawasaki Hiroaki , Yaguchi Tomoko , Kaul Sunil C. , Nakajima Motowo , Taira Kazunari
Published: March 2004

Background

Metastatic properties of tumors involve movement of cancerous cells from one place to another and tissue invasion. Metastatic cells have altered cell adhesion and movement that can be examined by in vitro chemotaxis assays. The Rho/ROCK/LIM kinase pathway is one of the major signaling pathways involved in tumor metastasis. It is involved in the regulation of the actin cytoskeleton. Using the randomized ribozyme library, we initially found that metastatic human fibrosarcoma cells harboring ribozyme specific for ROCK lose their metastatic properties. In this study, we have determined the effect of ribozymes specific for LIM kinase‐2 on metastatic and proliferative phenotypes of human fibrosarcoma cells.

Methods

We attempted to target LIM kinase‐2 (LIMK‐2) expression by hammerhead ribozymes (Rz) in human metastatic fibrosarcoma cells. An effective ribozyme was selected based on the expression analysis. Cells were stably transfected with Rz specifically effective for LIMK‐2 and were examined for metastatic and proliferative properties.

Results

Analyses of cellular phenotypes such as cell proliferation, cell migration and colony‐forming efficiency revealed that the suppression of LIMK‐2 expression in human fibrosarcoma cells limits their migration and dense colony‐forming efficiency without affecting cell proliferation rate or viability.

Conclusions

Specific targeting of metastatic and malignant properties of tumor cells by LIMK‐2 ribozyme may serve as an effective therapy for invasive tumors with minimum effect on the surrounding normal cells. Copyright © 2004 John Wiley & Sons, Ltd.
Lactosylated polyethylenimine for gene transfer into airway epithelial cells: role of the sugar moiety in cell delivery and intracellular trafficking of the complexes
Authors: Grosse Stéphanie , Aron Yolande , Honoré Isabelle , Thévenot Guiti , Danel Claire , Roche Annie‐Claude , Monsigny Michel , Fajac Isabelle
Published: March 2004

Background

As we have previously shown that lactosylated polyethylenimine (PEI) is the most efficient glycosylated PEI for gene transfer into human airway epithelial cells in primary culture, we have studied here the role of the lactose residue in the enhancement of gene transfer efficiency observed with lactosylated PEI as compared with unsubstituted PEI in immortalized (ΣCFTE29o‐ cells) and primary human airway epithelial cells.

Methods and results

After three transfections of 1 h performed daily, 60% of ΣCFTE29o‐ cells were transfected with lactosylated PEI, whereas 25% of cells were transfected with unsubstituted PEI (p < 0.05). Cell viability was 1.8‐fold greater with lactosylated PEI as compared with unsubstituted PEI (p < 0.05). As assessed by flow cytometry, the cellular uptake of lactosylated complexes was greater than that of complexes made with unsubstituted PEI (p < 0.05) and involved mostly a receptor‐mediated endocytosis. The study of the intracellular trafficking in airway epithelial cells of complexes showed an endosomal and lysosomal accumulation of lactosylated complexes. In the presence of a proton pump inhibitor, the level of lactosylated and unsubstituted PEI‐mediated gene expression was reduced more than 20‐fold, whereas the cell viability increased to almost 100%. For both complexes, a nuclear localization was observed for less than 5% of intracellular complexes.

Conclusions

Our results show that the greater gene transfer efficiency observed for lactosylated complexes may be attributed to a higher amount of lactosylated complexes incorporated by airway epithelial cells and a lower cytotoxicity that might be related to reduced endosomolytic properties. However, the lactose residues substituting the PEI did not promote the entry of the plasmid into the nucleus. Copyright © 2004 John Wiley & Sons, Ltd.
Biodistribution of the RD114/mammalian type D retrovirus receptor, RDR
Authors: Green Bronwyn J. , Lee C. Soon , Rasko John E. J.
Published: March 2004

Background

The limited expression of viral receptors on target cells is a recognized barrier to therapeutic gene transfer. Previous analysis of receptor expression has been performed using indirect methods due to a lack of receptor‐specific antibodies.

Methods

In this report we have used anti‐RDR antiserum to provide direct histochemical and flow cytometric analysis of the expression of RDR, which is the cognate receptor for RD114‐pseudotyped vectors as well as being a neutral amino acid transporter.

Results

RDR was present on a range of normal tissues with relevance to gene therapy including: colon, testis, ovary, bone marrow and skeletal muscle. It was also highly expressed on immature cells present in the squamous epithelia of skin, cervix, nasal mucosa, bronchus and tonsil. Of relevance to possible germline gene transfer, we demonstrated a lack of RDR expression on male or female germ cells. RDR expression on mature hemopoietic cell subsets showed up to 5‐fold variability between individuals within each lineage—with some individuals expressing low levels of RDR across all blood lineages. Both myeloid and monocytic lineages contained the highest fraction of cells expressing RDR, whereas lymphoid lineages showed the lowest. Coexpression of CD34 and RDR ranged from 2.04 to 0.44% in G‐CSF‐mobilized peripheral blood samples.

Conclusions

As a means to optimize gene transfer protocols, biodistribution studies such as these are fundamental to enable targeting of the virus receptor most abundantly expressed on relevant populations. The inter‐individual variation of receptor expression seen here also raises the possible requirement for tailor‐made gene therapy protocols. Copyright © 2004 John Wiley & Sons, Ltd.