bluuebunny/crossref_metadata_2025 huggingface dataset:
Generalized Hough transform to be extended as an affine‐invariant detector of arbitrary shapes
Authors: Kimura Akio , Watanabe Takashi
Published: May 2004

The problem of detecting an arbitrary curved shape from an image in an affine‐invariant form is an important basic research topic in image recognition. Although several conventional techniques for achieving this kind of detection goal have been proposed, none of them are able to deal with the following kinds of situations very well: (1) part of the contour of the shape to be detected is hidden due to overlap with another object and (2) breaks occur in the contour due to noise. Therefore, in this paper, the authors propose a new shape detection method that can even deal with these kinds of situations. First, to stably extract tangent information from a discontinuous contour shape that includes arbitrary curves, they propose a new tangent information extraction method based on arc fitting using a Hough transform. They then focus on the property that the “parallelism of straight lines is preserved under an affine transformation” and actively use tangent information at various contour point positions of the shape to extend Ballard's generalized Hough transform 1 as an affine‐invariant shape detection method. The authors performed evaluation experiments to verify the effectiveness of the proposed technique and obtained good detection results. © 2004 Wiley Periodicals, Inc. Electron Comm Jpn Pt 2, 87(6): 58–68, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjb.20095
Sampling correction in linkage analysis
Authors: Ginsburg Emil , Malkin Ida , Elston Robert C.
Published: May 2004

In a linkage analysis that requires the estimation of parameters other than the recombination fraction, we can construct a pedigree likelihood that leads to consistent parameter estimators if the sampling procedures are known. In particular, it is necessary to identify the subset of pedigree members “relevant to sampling” (RS), where by sampling we mean both pedigree ascertainment through a proband combination and the selective inclusion of the sampled pedigrees in the data that are analyzed. If both these procedures are independent of the marker phenotypes and the model of trait inheritance is known, then no sampling or ascertainment correction is needed to obtain a consistent estimator of the recombination fraction. Otherwise, the correction can be of two types: sampling‐model‐based, in which the ascertainment and inclusion procedures are modeled and used in the likelihood expression, or sampling‐model‐free, in which the data RS are “conditioned out” without any modeling of the sampling procedures. In either case, the pedigree proband sampling frame must be identified. © 2004 Wiley‐Liss, Inc.
Gamma radiation–induced apoptosis, G2 delay, and the risk of salivary and thyroid carcinomas—a preliminary report
Authors: Zheng Rong , Dahlstrom Kristina R. , Wei Qingyi , Sturgis Erich M.
Published: May 2004

Background.

While radiation has been the only well‐established risk factor for salivary and thyroid cancers, the exact mechanisms remain unknown. We hypothesized that individuals with altered apoptotic response to γ irradiation may be susceptible to salivary and thyroid cancers.

Methods.

We tested our hypothesis in a pilot case‐control study of 29 patients with neoplasms of the salivary and thyroid glands and 29 cancer‐free control subjects. Patients and control subjects were matched on age, sex, and ethnicity. In vitro γ radiation–induced apoptosis in lymphocytes was quantified utilizing the TUNEL assay and flow cytometry.

Results.

The mean apoptotic capacity was 13.55 ± 10.54 for control subjects, 5.75 ± 4.96 for patients with salivary gland carcinomas (p = .003), and 6.87 ± 4.45 for patients with thyroid carcinomas (p = .006). These differences were associated with a 10‐fold increased risk of salivary gland carcinoma (odds ratio [OR] = 10.71; 95% confidence interval [CI], 1.21–94.86) and a four‐fold increased risk of thyroid carcinoma (OR = 3.93; 95% CI, 0.90–17.08).

Conclusions.

Our data suggests that γ radiation–induced apoptosis may serve as a biomarker of genetic susceptibility to salivary and thyroid carcinoma, and further confirmatory studies with larger sample size are warranted. © 2004 Wiley Periodicals, Inc. Head Neck 26: 612–618, 2004
IL‐6 as an intracrine growth factor for renal carcinoma cell lines
Authors: Alberti Laurent , Thomachot Marie Cécile , Bachelot Thomas , Menetrier‐Caux Christine , Puisieux Isabelle , Blay Jean Yves
Published: May 2004

Interleukin‐6 (IL‐6) is produced at high levels by renal cell carcinoma cell lines. The molecular mechanisms involved in its possible role as an autocrine growth factor were investigated. IL‐6 and IL‐6 receptor expression was investigated in 8 renal cell carcinoma (RCC) cell lines. The modulation of RCC cell line proliferation by an anti‐IL‐6 Ab, an IL‐6 antisense oligonucleotide (ASON) directed against the second exon of IL‐6 and cytokines inhibiting IL‐6 production (IL‐4 and IL‐13) was investigated. All 8 RCC cell lines expressed IL‐6 mRNA, produced IL‐6 and expressed the soluble and membrane‐bound gp130 chain of IL‐6 receptor. The gp80 chain of IL‐6 receptor was undetectable at the surface of the 8 RCC cell lines tested, while the soluble form of gp80 was detectable in the supernatant of one of these cell lines. The addition of a blocking IL‐6 Ab did not inhibit the proliferation of any of the 8 RCC cell lines. In contrast, IL‐6 ASON inhibited specifically IL‐6 production and the proliferation of all RCC cell lines. Exogenous IL‐6 failed to restore RCC cell line proliferation blocked by ASON, indicating that IL‐6 acts through an intracrine loop in RCC cell lines. IL‐13 and IL‐4 inhibited the proliferation of 7 of the 8 cell lines without interfering with IL‐6 or IL‐6 receptor expression. IL‐6 ASON inhibited the proliferation of the 8 RCC cell lines tested additively with IL‐4 or IL‐13. IL‐6 is an intracrine growth factor in renal cell carcinoma cell lines. © 2004 Wiley‐Liss, Inc.
Gene transfer to cervical cancer with fiber‐modified adenoviruses
Authors: Rein Daniel T. , Breidenbach Martina , Wu Hongju , Han Tie , Haviv Yosef S. , Wang Minghui , Kirby Tyler O. , Kawakami Yosuke , Dall Peter , Alvarez Ronald D. , Curiel David T.
Published: May 2004

Successful adenoviral (Ad) vector–mediated strategies for cancer gene therapy mandate gene‐delivery systems that are capable of achieving efficient gene delivery in vivo. In many cancer types, in vivo gene‐transfer efficiency remains limited due to the low or highly variable expression of the primary Ad receptor, the coxsackie Ad receptor (CAR). In this study, we evaluated the expression of CAR on cervical cancer cells as well as CAR‐independent targeting strategies to integrins (Ad5.RGD), heparan sulfate proteoglycans (Ad5.pK7) or both (Ad5.RGD.pK7). We used a panel of established cervical cancer cell lines and primary cervical cancer cells isolated from patients to quantify the expression of CAR mRNA and to evaluate the gene‐transfer efficiency of fiber‐modified Ads. Of the fiber‐modified vectors, Ad5.pK7 and Ad5.RGD.pK7 displayed significantly enhanced gene‐transfer efficiency in vitro. Gene‐delivery efficiency in vivo was evaluated using an s.c. cervical cancer mouse model. Ad5.RGD.pK7 significantly improves tumor targeting in vivo, resulting in a significantly improved tumor/liver ratio in mice. Our results suggest that the double‐modified Ad5.RGD.pk7 vector enhances gene transfer to clinically relevant cervical cancer substrates, while the infectivity of nontarget cells in the mouse is not increased and comparable to Ad5. The fiber‐modified virus described here can help achieve higher clinical efficacy of cervical cancer gene therapy. © 2004 Wiley‐Liss, Inc.
Identification of a small peptide that inhibits the phosphorylation of ErbB2 and proliferation of ErbB2 overexpressing breast cancer cells
Authors: Pero Stephanie C. , Shukla Girja S. , Armstrong Amy L. , Peterson Daniel , Fuller Susan P. , Godin Katherine , Kingsley‐Richards Sarah L. , Weaver Donald L. , Bond Jeffrey , Krag David N.
Published: May 2004

ErbB2 is overexpressed in approximately 30% of breast cancer patients with a correlation to poor prognosis. ErbB2 has been identified as a useful receptor for molecular targeting. A cyclic 20 amino acid phage display random peptide library was constructed using the fUSE5 gene III system. The library was panned against 2 different purified forms of the external domain of ErbB2. This resulted in the identification of several ErbB2‐binding phage clones with variable binding to different ErbB2 preparations. One clone (EC‐1) bound all preparations of ErbB2 including live cells and fresh frozen human breast cancer specimens. The synthetic peptide based on the deduced sequence of the EC‐1 clone and its biotin‐conjugated form retained binding affinity for purified ErbB2 and ErbB2 overexpressing cell lysates. EC‐1 peptide was able to effectively inhibit the phosphorylation of ErbB2 on residues Y1248 and Y877 in a dose‐ and time‐dependent manner. Furthermore, EC‐1 peptide selectively inhibits the proliferation of ErbB2 overexpressing breast cancer cells. The linear portion of the cyclic EC‐1 peptide was shown to be essential for binding ErbB2. In addition, 4 biased phage libraries were constructed allowing 4 different regions of the EC‐1 peptide to have random sequence. Screening these EC‐1 biased libraries did not result in higher affinity peptides but did demonstrate the importance of amino acids at position 1–4 on the N‐terminal flanking arm and 11–15 within the cyclic ring. Interestingly, EC‐1 contains homologous motifs with known ErbB receptor family ligands. We have identified a small peptide that binds to the extracellular domain of ErbB2, inhibits ErbB2 autophosphorylation and inhibits the proliferation of ErbB2 overexpressing cells. This supports the notion that small peptides can bind to targets important in cancer therapy even if a target does not have a natural ligand. Continuing research with this peptide includes increasing its affinity to ErbB2, evaluation of pharmacokinetics and evaluation of anti‐proliferative effects with conjugate anti‐cancer agents. © 2004 Wiley‐Liss, Inc.
Frequent amplification and overexpression of CCND1 in male breast cancer
Authors: Bärlund Maarit , Kuukasjärvi Tuula , Syrjäkoski Kirsi , Auvinen Anssi , Kallioniemi Anne
Published: May 2004

Genetic events underlying the pathogenesis of breast cancer have been studied extensively and several clinically significant markers have been identified. For example, amplification and overexpression of the ERBB2 oncogene is associated with poor prognosis in breast cancer and ERBB2 serves as a target for antibody‐based therapy. Current knowledge on the pathogenesis of male breast cancer (MBC) is limited. The purpose of our study was to investigate the potential relevance of a series of genes known to be amplified in female breast cancer (FBC) in a the development and pathogenesis of MBC. To this end, we applied fluorescence in situ hybridization and immunohistochemistry to the analysis of 128 breast tumors from males. Amplification of ERBB2, MYC, PPM1D and ZNF217 was detected rarely (1–2% of tumors) indicating a considerably lower amplification frequency than in FBC. CCND1 amplification was observed in 12% of cases, being in good concordance with findings from FBC. In addition, CCND1 overexpression was detected in 63% of tumors and was associated with ER positivity (p < 0.0001). Our results indicate distinct differences in the genetic basis of MBC and FBC and suggest that marked differences exist in the pathogenesis of these diseases. The lack of ERBB2 involvement was especially unexpected and implies that ERBB2‐targeted therapies are unlikely to be beneficial in MBC. Furthermore, the high frequency of hormone receptor positivity and the association between ER positivity and CCND1 overexpression supports the notion that hormonal regulation is likely to be essential for the development of MBC. © 2004 Wiley‐Liss, Inc.
In vitro blockade of receptor activator of nuclear factor‐κB ligand prevents osteoclastogenesis induced by neuroblastoma cells
Authors: Granchi Donatella , Amato Ilaria , Battistelli Luca , Avnet Sofia , Capaccioli Sergio , Papucci Laura , Donnini Martino , Pellacani Andrea , Brandi Maria Luisa , Giunti Armando , Baldini Nicola
Published: May 2004

Proliferation and differentiation of osteoclasts are regulated by a cytokine system that includes RANKL, which binds 2 receptors: RANK, which activates osteoclast differentiation, and osteoprotegerin (OPG), a decoy receptor that limits RANKL action. We investigated the role of the OPG/RANKL/RANK network in the pathogenesis of skeletal metastasis in neuroblastoma. Four different neuroblastoma cell lines (NB100, CHP212, SH‐SY5Y, SJ‐NK‐P) showed a large amount of OPG and RANKL transcripts. Soluble RANKL was detectable in all cell lines, but poor release of OPG was observed. SH‐SY5Y showed the lowest OPG‐to‐RANKL ratio and promoted osteoclastic differentiation of FLG29.1 and peripheral mononuclear cells, inducing expression of the osteoclast markers RANK, c‐src, c‐fos, cathepsin‐K and TRAP. SJ‐N‐KP, which released both OPG and RANKL, did not show the same capability. OPG, neutralizing anti‐RANKL antibody and antisense oligonucleotides were evaluated for their ability to inhibit RANKL activity. The neutralizing antibody hampered osteoclastic differentiation by blocking both the juxtacrine and the paracrine activity of RANKL. Our findings confirm that neuroblastoma cells induce osteoclastogenesis via RANKL and suggest that the RANKL expression associated with lack of the decoy receptor OPG could be a peculiarity of some tumors that makes them able to induce metastatic osteolysis. Moreover, our results suggest that RANKL could be a relevant target in the adjuvant therapy of bone metastatic neuroblastoma as proper neutralization revokes completely osteoclastic differentiation. © 2004 Wiley‐Liss, Inc.
Non‐self‐discrimination as a driving concept in the identification of an immunodominant HMW‐MAA epitopic peptide sequence by autoantibodies from melanoma cancer patients
Authors: Dummer Reinhard , Mittelman Abraham , Fanizzi Francesco P. , Lucchese Guglielmo , Willers Jörg , Kanduc Darja
Published: May 2004

We analyzed the sera of patients with melanoma to define the human humoral autoantibody profile towards HMW‐MAA. Computational proteome scanning using the non‐self‐discrimination principle as a guide led to the individuation of the low‐similarity HMW‐MAA781–789RATVWMLRL peptide fragment as an immunodominant B‐cell epitope. Linear B‐cell determinant individuation was experimentally validated by dot blot immunoassay and NMR spectroscopy analysis. Regulation of physiologic self‐reactivity by the non‐self‐discrimination principle is discussed. © 2004 Wiley‐Liss, Inc.
Clinical relevance of amphiregulin and VEGF in primary breast cancers
Authors: Desruisseau Sylvie , Palmari Jaqueline , Giusti Corinne , Romain Sylvie , Martin Pierre‐Marie , Berthois Yolande
Published: May 2004

The characterization of novel prognostic markers in breast cancer is necessary to improve the identification of high‐risk populations. In our study, the prognostic significance of VEGF and amphiregulin (AR) was investigated and compared to conventional prognostic factors in primary breast cancers. The analysis was performed using enzyme‐linked immuno‐assay in a series of 193 patients, and univariate and multivariate analysis were performed in the overall population as well as in pre‐ and post‐menopausal patients subdivided in node‐negative (N−) and node‐positive (N+) subsets. AR (median, 44.8 pg/mg protein) appeared strongly correlated with progesterone receptors (PgR) (p = 0.0018) in the premenopausal N+ population, and with uPA (p= 0.020) and VEGF (p= 0.0053) in the postmenopausal/N+ patients. Despite these attractive data, AR expression was not significant for recurrence or survival outcome. Data revealed strong correlation between VEGF and uPA, and PAI‐1, in the N+ population. Moreover, patients with high VEGF levels displayed poor outcome, with an increased risk for N+ subset. These data were confirmed by multivariate analysis that presented histologic grade (HR, 10.55, p = 0.001) and VEGF (HR, 3.89, p = 0.03) as the prominent prognostic markers for overall survival for the N+ population. Furthermore, infiltrating ductal carcinomas (IDC) were shown to express higher levels of both uPA (p < 0.0001) and VEGF (p = 0.002) than intralobular carcinomas. This retrospective study reinforces the pejorative biological role of VEGF in the progression of breast tumors. Our data also suggest that VEGF and uPA might play particular role in the biology and progression of IDC. © 2004 Wiley‐Liss, Inc.
Aerodigestive and gastrointestinal tract cancers and exposure to crocidolite (blue asbestos): Incidence and mortality among former crocidolite workers
Authors: Reid Alison , Ambrosini Gina , de Klerk Nicholas , Fritschi Lin , Musk Bill
Published: May 2004

The objective of this article was to assess the association between the incidence and mortality from aerodigestive cancers and exposure to crocidolite (blue asbestos). Our study is a cohort study of former workers of the now‐defunct crocidolite mining and milling operation at Wittenoom, Western Australia, who have been followed up since 1979 and on whom asbestos exposure and smoking information was known. Standardised mortality and incidence rates were used to compare former workers with the Western Australian male population. Cases were matched with up to 10 randomly assigned controls, and conditional logistic regression was used to examine the relationship between asbestos exposure, smoking status and cancer incidence. There were 129 incident cases from all cancers of interest and 57 deaths. Former workers had a significantly higher risk of mortality from upper aerodigestive cancers than the Western Australian male population. The incidence of upper and lower aerodigestive cancers was higher in the Wittenoom cohort but not significantly so. Cumulative exposure to asbestos did not appear to be associated with the incidence of stomach cancer, colorectal cancer or upper aerodigestive cancers. Smoking status was strongly associated with the incidence of upper aerodigestive cancers, with current smokers experiencing the greatest risk. Our study with longer and more complete follow‐up, smoking information and a stronger study design does not show an association between cumulative asbestos exposure and stomach cancer or other gastrointestinal cancers. The excess mortality from upper aerodigestive cancers seen in this cohort of former asbestos workers compared to the Western Australian male population does not appear to be associated with exposure to crocidolite. © 2004 Wiley‐Liss, Inc.
Breast cancer metastatic potential: Correlation with increased heterotypic gap junctional intercellular communication between breast cancer cells and osteoblastic cells
Authors: Kapoor Preeti , Saunders Marnie M. , Li Zhongyong , Zhou Zhiyi , Sheaffer Nate , Kunze Elaine L. , Samant Rajeev S. , Welch Danny R. , Donahue Henry J.
Published: May 2004

The breast cancer metastasis‐suppressor gene BRMS1 is downregulated in metastatic breast cancer cells. Previous reports have shown restoration of gap junctional intercellular communication (GJIC) in the metastatic human breast carcinoma cell line MDA‐MB‐435 (435) transfected with BRMS1 cDNA. Metastasis, to a large extent in most breast cancers, occurs to bone. However, the reason for this preferential metastasis is not known. We explored cell‐to‐cell communication between 435 carcinoma cells and a human osteoblastic cell line, hFOB1.19, to determine whether carcinoma cells can form gap junctions with bone cells and to explore the role of these heterotypic gap junctions and the BRMS1 gene in breast cancer metastasis to bone. 435 cells displayed greater cell‐to‐cell communication with hFOB 1.19 cells than with themselves. Transfection of BRMS1 into 435 cells increased homotypic gap junctional communication but did not significantly affect heterotypic communication with hFOBs. However, heterotypic communication of BRMS1 transfectants with hFOB cells was reduced relative to homotypic communication. In contrast, parental 435 cells displayed greater heterotypic communication with hFOBs relative to homotypic communication. Our results suggest that there are differences in the relative homotypic and heterotypic GJIC of metastasis‐capable and ‐suppressed cell lines. © 2004 Wiley‐Liss, Inc.
Childbirth and breast cancer prognosis
Authors: Rosenberg Lena , Thalib Lukman , Adami Hans‐Olov , Hall Per
Published: May 2004

Although certain risk factors for breast cancer incidence may also effect survival, findings have been inconsistent and the long‐term role of childbirth is unknown. We studied the influence of number and timing of births on breast cancer prognosis prospectively. From 1958 to 1997, altogether 32,003 women, born 1932 or later, were notified to the Swedish Cancer Registry due to a primary invasive breast cancer. We obtained information on dates of all childbirths and achieved complete follow‐up through 1997 by means of linkage to other nation‐wide databases. Proportional hazards analyses were used to compute crude and multivariate hazard ratios (HR) with 95% confidence intervals (CI) of dying from breast cancer. We found a successively worse prognosis for women with a shorter delay between their last birth and breast cancer diagnosis (p for trend <0.0001). Compared to women with their last birth more than 10 years before diagnosis, the multivariate HR of breast cancer death was 1.39 (95% CI 1.17–1.67) for those with breast cancer diagnosis in the 3rd year after last birth and 1.72 (CI 95% 1.42–2.09) for those with diagnosis within 1 year after last birth. This adverse effect on prognosis of childbirth persisted beyond 10 years among women with a first birth before the age of 20 years. A pregnancy has marked adverse effects on the prognosis of a breast cancer diagnosed within 10 years after delivery. These findings suggest that pregnancy influences tumor biology. © 2004 Wiley‐Liss, Inc.
High BRAF mutation frequency does not characterize all melanocytic tumor types
Authors: Saldanha Gerald , Purnell David , Fletcher Alan , Potter Linda , Gillies Angela , Pringle J. Howard
Published: May 2004

Cutaneous melanoma (CM) is the most lethal form of skin cancer. Along with some benign melanocytic tumors, the majority shows BRAF or NRAS mutation, but it is not known whether these are essential to all forms of melanocytic neoplasia. We screened 79 melanocytic tumors of different types for BRAF and NRAS mutations and looked at MAPK pathway activity using immunohistochemistry in a subset. Significant differences in BRAF exon 15 mutation frequency were found: 14/16 (87.5%) in common acquired naevi (CANs), 9/12 (75%) in CMs, 0/26 in Spitz naevi and 3/25 (12%) in blue naevi (p < 0.01). We looked at whether Spitz and blue naevi showed a compensatory increase in BRAF exon 11 and/or NRAS exons 1 and 2 mutations to account for the low BRAF exon 15 mutation frequency. NRAS mutations were found in only 1/16 (6.3%) Spitz naevi and 0/15 blue naevi. In addition, NRAS mutations were found in 2/11 (18.2%) CANs and 3/12 (25%) CMs. None of the tumors showed BRAF exon 11 mutations. Despite their low combined BRAF and NRAS mutation frequency, Spitz naevi showed strong MAPK pathway activation as measured by cytoplasmic expression of dually phosphorylated ERK1/2, while blue naevi had weak pathway activation. We conclude that BRAF and NRAS mutations are not necessary for melanocytic tumor development and that some types of tumor must arise by alternative mechanisms. © 2004 Wiley‐Liss, Inc.
PIBF (progesterone induced blocking factor) is overexpressed in highly proliferating cells and associated with the centrosome
Authors: Lachmann Margit , Gelbmann Dieter , Kálmán Endre , Polgár Beata , Buschle Michael , von Gabain Alexander , Szekeres‐Barthó Júlia , Nagy Eszter
Published: May 2004

PIBF was previously identified as a 34 kDa immunomodulatory molecule secreted by pregnancy lymphocytes and is thought to play a crucial role in preventing rejection of the embryo by the maternal immune response. Recent data suggested that PIBF protein was also expressed by the progesterone receptor (PR) positive MCF‐7 breast tumor cell line. Therefore our study was designed to analyze the expression of PIBF in malignant cell lines and primary tumors both at the mRNA and protein levels. RNA expression analyses of several human cell lines with different tissue origin and paired human tumor/normal tissues, as well as of several PR+ and PR− breast tumors revealed that PIBF mRNA was overexpressed in highly proliferating cells independent of the presence of PR. In addition to the full‐length PIBF mRNA encoding for a 90 kDa protein, several alternatively spliced species were detected, all resulting from perfect exon skipping. The most frequently identified splice variant is predicted to encode for an approximately 35 kDa protein. Immunofluorescence microscopy revealed a centrosomal localization for the full‐length PIBF, while the 35 kDa form showed a diffuse cytoplasmic staining. These data, together with the identification of the PIBF gene in the chromosomal region associated with breast cancer susceptibility, reveal a strong parallel with known tumor suppressor proteins, such as BRCA1 and p53 having the same centrosomal localization. Given the notion that a number of proteins shown to be involved in tumorigenesis are associated with the centrosome and disturbed centrosome function causes unequal segregation of chromosomes, studies to evaluate whether or not PIBF that is highly expressed in tumors is directly involved in tumorigenesis are thus warranted. © 2004 Wiley‐Liss, Inc.
Amount of DNA in plasma and cancer risk: A prospective study
Authors: Gormally Emmanuelle , Hainaut Pierre , Caboux Elodie , Airoldi Luisa , Autrup Herman , Malaveille Christian , Dunning Alison , Garte Seymour , Matullo Giuseppe , Overvad Kim , Tjonneland Anne , Clavel‐Chapelon Francoise , Boffetta Paolo , Boeing Heiner , Trichopoulou Antonia , Palli Domenico , Krogh Vittorio , Tumino Rosario , Panico Salvatore , Bueno‐De‐Mesquita H. Bas , Peeters Petra H. , Lund Eiliv , Gonzalez Carlos A. , Martinez Carmen , Dorronsoro Miren , Barricarte Aurelio , Tormo M. Jose , Quiros J. Ramón , Berglund Goran , Hallmans Goran , Day Nicholas E. , Key Timothy J. , Veglia Fabrizio , Peluso Marco , Norat Teresa , Saracci Rodolfo , Kaaks Rudolf , Riboli Elio , Vineis Paolo
Published: May 2004

Levels of plasma DNA concentrations in cancer patients have been shown to be higher than the plasma DNA concentrations found in healthy subjects. The value of plasma DNA levels for development of neoplastic or pulmonary disease was evaluated in a large prospective study. Plasma samples (n = 1,184) were analyzed from 776 controls, 359 cases of cancer (lung, bladder, oral cavity, pharynx, larynx, leukemia) and 49 deaths from chronic obstructive pulmonary disease (COPD), including never smokers and ex‐smokers, from 9 countries across Europe. The amount of plasma DNA was variable across the European Prospective Investigation into Cancer and Nutrition (EPIC) centers. High DNA concentrations in some centers might be due to the type of population recruited and/or the treatment of the samples. An elevated and statistically significant odds ratio (OR) was found for COPD deaths (OR = 2.53; 95% CI = 1.06–6.02), while nonsignificant increased ORs were present for oral cancers, cancers of the pharynx and larynx and leukemia. When the analyses were stratified by time since recruitment (below or above 36 months), the increased ORs were limited to the more recent period of recruitment, i.e., a time elapsed between blood drawing and disease onset lower than 36 months. This was particularly true for COPD deaths (OR = 12.7; 95% CI = 1.57–103) and leukemia (OR = 2.37; 95% CI = 1.20–4.67). © 2004 Wiley‐Liss, Inc.
Increased expression of amyloid precursor protein in oral squamous cell carcinoma
Authors: Ko Shun‐Yao , Lin Shu‐Chun , Chang Kuo‐Wei , Wong Yong‐Kie , Liu Chung‐Ji , Chi Chin‐Wen , Liu Tsung‐Yun
Published: May 2004

In our previous study, we identified amyloid precursor protein (APP) in an oral squamous cell carcinoma (OSCC)‐enriching subtractive hybridization library. Our present study attempts to define the significance of APP expression in the genesis of OSCC. RT‐PCR analysis showed increase in APP mRNA expression for more than 2‐fold in 76% of OSCC (n = 55) relative to corresponding non‐cancerous matched tissues (NCMT). The majority of esophageal SCCs also had increase in APP mRNA expression. OSCC patients exhibiting increase in APP mRNA expression had significantly lower survival rate compared to patients exhibiting the opposite status. Western blotting analysis identified APP751 and APP770 as the major APP isoforms in oral keratinocytes. A high correlation between mRNA and protein expressions of APP was noted in OSCC/NCMT pairs. Immunohistochemistry further showed a remarkable increase of APP in OSCC tissue relative to NCMT. Treatment with an antisense oligonucleotide against APP reduced cellular and secreted APP as well as growth in an OSCC cell line. Our study provides novel clues that APP expression is involved in the proliferation and carcinogenesis of OSCC. Correlated with such pathogenesis was the survival of its victims. The degree of APP expression could serve as an invaluable marker for oral carcinogenesis. © 2004 Wiley‐Liss, Inc.
Gene expression profiles in breast tumors regarding the presence or absence of estrogen and progesterone receptors
Authors: Nagai Maria Aparecida , da Rós Nancy , Neto Mário Mourão , de Faria Junior Silvio Rodrigues , Brentani Maria Mitzi , Hirata Roberto , Neves Eduardo Jordão
Published: May 2004

Estrogen acts via its receptor (ER) to stimulate cell growth and differentiation in the mammary gland. ER and progesterone receptor (PR), which is regulated by estrogen via ER, have been used as prognostic markers in clinical management of breast cancer patients. Patients with ER− breast tumors have a poorer prognosis than patients with ER+ tumors. The aim of the present study was the identification of tumor‐associated genes differentially expressed in breast tumors regarding the presence or absence of ER and PR hybridized with cDNA microarrays containing 4,500 tumor‐derived expressed sequence tags generated using the ORESTES technique. Samples of human primary breast carcinomas from 38 patients were analyzed. The experiments were performed in triplicates and data from each element were acquired by phosphoimage scanning. Data acquisition was performed using the ArrayVision software. After normalization statistical analysis was applied. In a preliminary analysis, 98 differentially expressed transcripts were identified, 46 were found to be more expressed in ER+/PR+ and 52 were found to be more expressed in ER−/PR− breast tumors. The biochemical functions of the genes in the reported expression profile are diverse and include metabolic enzymes, protein kinases, helicases, transcription factors, cell cycle regulators and apoptotic factors. ER−/PR− breast tumors displayed increased levels of transcripts of genes associated with neurodegeneration and genes associated with proliferation were found in ER+/PR+ tumors. © 2004 Wiley‐Liss, Inc.
Use of adenoviruses encoding CD40L or IL‐2 against B cell lymphoma
Authors: Meziane El Kahina , Bhattacharyya Tapan , Armstrong Anne C. , Qian Cheng , Hawkins Robert E. , Stern Peter L. , Dermime Said
Published: May 2004

Some B cell lymphomas lack important costimulatory properties that could prevent them from being used as cell based vaccines. Infection of A20 B lymphoma cells with a replication‐defective adenovirus encoding murine (m) CD40L, but not mIL‐2, produces an antigen presentation phenotype with upregulation of MHC Class I/II, induction of B7‐1/2 molecules and production of MIL‐12 and MIP‐1α. Subcutaneous vaccination with irradiated Ad‐mCD40L‐infected‐ or Ad‐mIL‐2‐infected‐A20 cells generated A20‐specific CD8+ T cell responses and cross reactive A20 Ig antibodies. Only vaccination with Ad‐mCD40L‐infected A20 cells produced a significant delay in tumor growth and long‐term survival (p = 0.0039). Stronger protective immunity to A20 challenge was generated by intravenous priming with A20 cells infected with Ad‐mCD40L, Ad‐mIL‐2 or their combination followed by a boost immunization with A20 cells activated with syngeneic fibroblasts expressing CD40L. Compared to Ad‐LacZ‐infected A20 priming, the combination priming was most effective followed by Ad‐mCD40L and Ad‐mIL‐2 (p = 0.0027, p = 0.0027, p = 0.0163 respectively). Significant A20‐specific CD8+ T cell‐mediated cytotoxicity was only demonstrated in splenocytes from these groups of vaccinated animals. By contrast, ELISPOT assay of splenocytes from all A20 prime/boosted vaccinated groups demonstrated increases in γ‐interferon release by T cells elicited by in vitro stimulation either with A20 cells or another syngeneic 2PK‐3 lymphoma, indicating the presence of cross reactive immunity. Similarly anti‐A20 immunoglobulin antibodies generated after vaccination were not necessarily A20 idiotype‐specific. Direct therapy of pre‐established tumors was achieved with the combination of Ad‐mCD40L and Ad‐mIL‐2 given at Days 4 and 8 at the tumor site with a significant long‐term survival of 85% of tumor‐bearing mice (p = 0.0001). Our study strongly supports the use of Ad‐CD40L and Ad‐IL‐2 combination therapy for the treatment of patients with B cell lymphoma. © 2004 Wiley‐Liss, Inc.
Inhibition of growth and survival of human head and neck squamous cell carcinoma cells by curcumin via modulation of nuclear factor‐κB signaling
Authors: Aggarwal Sita , Takada Yasunari , Singh Sujay , Myers Jeffrey N. , Aggarwal Bharat B.
Published: May 2004

Increased expression of proinflammatory and proangiogenic factors are associated with aggressive tumor growth and decreased survival of patients with head and neck squamous cell carcinoma (HNSCC). In as much as genes that are regulated by nuclear factor NF‐κB suppress apoptosis, induce proliferation, and mediate inflammation, angiogenesis and tumor metastasis, agents that suppress NF‐κB activation have potential as treatment for various cancers including HNSCC. We demonstrate that all HNSCC cell lines expressed constitutively active NF‐κB and IκBα kinase (IKK), which is needed for NF‐κB activation. Treatment of MDA 686LN cells with curcumin (diferuloylmethane), a pharmacologically safe chemopreventive agent, inhibited NF‐κB activation through abrogation of IKK. As a result expression of various cell survival and cell proliferative genes including Bcl‐2, cyclin D1, IL‐6, COX‐2 and MMP‐9 was suppressed. This, in turn, inhibits proliferation of all HNSCC cell lines, arrests cell cycle in G1/S phase (MDA 686LN) and induces apoptosis as indicated by upstream and downstream caspase activation, PARP cleavage, annexin V staining in MDA 686LN cells. Suppression of NF‐κB by cell‐permeable p65‐based peptide and NBD peptide also inhibited the proliferation and induced apoptosis in these cells. Our results indicate that curcumin is a potent inhibitor of cell proliferation and an inducer of apoptosis in HNSCC through suppression of IKK‐mediated NF‐κB activation and of NF‐κB‐regulated gene expression. © 2004 Wiley‐Liss, Inc.
Treatment with siRNA and antisense oligonucleotides targeted to HIF‐1α induced apoptosis in human tongue squamous cell carcinomas
Authors: Zhang Qunzhou , Zhang Zuo‐Feng , Rao Jian Y. , Sato J. Denry , Brown Jimmy , Messadi Diana V. , Le Anh D.
Published: May 2004

Overexpression of hypoxia inducible factor‐1α (HIF‐1α) in cancers has been correlated to a more aggressive tumor phenotype. We investigated the effect of HIF‐1α knockout on the in vitro survival and death of human tongue squamous cell carcinomas (SCC‐4 and SCC‐9). Under normoxic condition, a basal level of HIF‐1α protein was constitutively expressed in SCC‐9 cells, albeit an undetectable level of HIF‐1α messages. Exposure to hypoxia induced only a transient increase in mRNA transcript but a prolonged elevation of HIF‐1α protein and its immediate downstream target gene product, VEGF. Under normoxic or hypoxic conditions, treatment of SCC‐9 cells with AS‐HIF‐1α ODN suppressed both constitutive and hypoxia‐induced HIF‐1α expression at both mRNA and protein levels. Knockout of HIF‐1α gene expression via either AS‐HIF‐1α ODN or siRNA (siRNAHIF‐1α) treatment resulted in inhibition of cell proliferation and induced apoptosis in SCC‐4 and SCC‐9 cells. We also demonstrated that exposure of SCC‐9 cells to hypoxia led to a time‐dependent increase in the expression of bcl‐2 and IAP‐2, but not p53. The attenuated levels of bcl‐2 and IAP‐2, and the enhanced activity of caspase‐3 after treatment with AS‐HIF‐1α ODN may contribute partly to the effects of HIF‐1α blockade on SCC‐9 cell death. Collectively, our data suggest that a constitutive or hypoxia‐induced expression of HIF‐1α in SCC‐9 and SCC‐4 cells is sufficient to confer target genes expression essential for tumor proliferation and survival. As a result, interfering with HIF‐1α pathways by antisense or siRNA strategy may provide a therapeutic target for human tongue squamous cell carcinomas. © 2004 Wiley‐Liss, Inc.
Addition of integrin binding sequence to a mutant human endostatin improves inhibition of tumor growth
Authors: Yokoyama Yumi , Ramakrishnan S.
Published: May 2004

Tumor vasculatures express high levels of αVβ3/αVβ5 and α5β1 integrins. Consequently, peptides containing the RGD (Arg‐Gly‐Asp) sequence, which is present in ligands of integrins, is effective in targeting therapeutic reagents to tumor vascular endothelium. In our study, we investigated whether the biologic activity of endostatin can be enhanced by the addition of an integrin targeting sequence. RGD sequence was added to either the amino or carboxyl terminus of endostatin containing a point mutation, P125A‐endostatin. Earlier we have shown that the P125A mutation did not affect the biologic activity of endostatin but in fact had better antiangiogenic activity when compared to the native molecule. Further modification of P125A‐endostatin with the RGD motif showed specific and increased binding to endothelial cells, and the increased binding coincided with improved antiangiogenic properties. Both amino and carboxyl terminal RGD‐modification of P125A‐endostatin resulted in greater inhibition of endothelial cell migration and proliferation. RGD modification increased tumor localization without affecting the circulatory half‐life of P125A‐endostatin, and RGD‐modified P125A‐endostatin was found to be more effective when compared to the P125A‐endostatin in inhibiting ovarian and colon cancer growth in athymic mice. Complete inhibition of ovarian tumor growth was observed when P125A‐endostatin‐RGD was encapsulated into alginate beads. These studies demonstrate that addition of a vascular targeting sequence can enhance the biologic activity of an antiangiogenic molecule. © 2004 Wiley‐Liss, Inc.
Association of DNA repair gene XRCC1 polymorphisms with head and neck cancer in Korean population
Authors: Tae Kyung , Lee Hyung Seok , Park Bum Jung , Park Chul Won , Kim Kyung Rae , Cho Hye Young , Kim Lyoung Hyo , Park Byung Lae , Shin Hyoung Doo
Published: May 2004

Squamous cell carcinoma of the head and neck (SCCHN), which is relatively prevalent in Korea, is believed to be induced by environmental carcinogens and host genetic factors. Accumulating evidence has shown that genetic differences in DNA repair capacity resulting from genetic polymorphism influence the risk of environmental carcinogenesis. We therefore examined the associations of genetic polymorphisms in the DNA repair genes XRCC1 with the risk of SCCHN in a Korean population (hospital‐based, case‐control study; 147 cases and 168 controls). Three known polymorphisms in the XRCC1 gene were genotyped: R194W(C>T) in exon 6, R280H(G>A) in exon 9 and R399G(G>A) in exon 10. Although no significant associations were apparent with R280H(G>A) and R399G(G>A), a highly significant association (p = 0.0005) of R194W(C>T) with the increased risk (OR = 2.61; 95% CI 1.53–4.46) of SCCHN was detected among patients and normal controls under dominant model. The frequency of minor allele‐containing genotypes (TT and CT) was much higher in SCCHN patients (51.8%) compared to that in normal controls (30.3%) (p = 0. 0005). When considering a relatively small number of cases (n = 147) and controls (n = 168) in our study, larger studies are needed to validate the genetic effects of XRCC1 polymorphisms in Asian populations. In conclusion, the result from our study provides additional evidence of an association of the XRCC1 polymorphism (Arg194Trp) with SCCHN as markers of genetic susceptibility in the Korean population. © 2004 Wiley‐Liss, Inc.
Suppression of malignant growth of human breast cancer cells by ectopic expression of integrin‐linked kinase
Authors: Chen Ping , Shen Wei‐Zhen , Karnik Pratima
Published: May 2004

Allelic loss at the short arm of chromosome 11 is one of the most common and potent events in the progression and metastasis of breast cancer. Here, we present evidence that the Integrin‐Linked Kinase (ILK) gene maps to the commonly deleted chromosome 11p15.5 and suppresses malignant growth of human breast cancer cells both in vitro and in vivo. ILK is expressed in normal breast tissue but is downregulated in metastatic breast cancer cell lines and in advanced breast cancers. Transfection of wild‐type ILK into the MDA‐MB‐435 mammary carcinoma cells potently suppressed their growth and invasiveness in vitro and reduced the cells' ability to induce tumors and metastasize in athymic nude mice. Conversely, expression of the ankyrin repeat or catalytic domain mutants of ILK failed to suppress the growth of these cells. Growth suppression by ILK is not due to apoptosis but is mediated by its ability to block cell‐cycle progression in the G1 phase and by modulating the levels of integrins. These findings directly demonstrate that ILK deficiency facilitates neoplastic growth and invasion and suggest a novel role for the ILK gene in the suppression of tumor metastasis. © 2004 Wiley‐Liss, Inc.
Poly(ADP‐ribosyl)ation inhibitors: Promising drug candidates for a wide variety of pathophysiologic conditions
Authors: Beneke Sascha , Diefenbach Joerg , Bürkle Alexander
Published: May 2004

Poly(ADP‐ribose) polymerases are involved in many aspects of regulation of cellular functions. Using NAD+ as a substrate, they catalyse the covalent transfer of ADP‐ribose units onto several acceptor proteins to form a branched ADP‐ribose polymer. The best characterised and first discovered member of this multiprotein family is PARP‐1. Its catalytic activity is markedly stimulated upon binding to DNA strand interruptions, and the resulting polymer is thought to function in chromatin relaxation as well as in signalling the presence of damage to DNA repair complexes and in regulating enzyme activities. Moderate activation of PARP‐1 facilitates the efficient repair of DNA damage arising from monofunctional alkylating agents, reactive oxygen species or ionising radiation, but severe genotoxic stress leads to rapid energy consumption and subsequently to necrotic cell death. The latter aspect of PARP‐1 activity has been implicated in the pathogenesis of various clinical conditions such as shock, ischaemia‐reperfusion and diabetes. Inhibition of ADP‐ribose polymer formation has been shown to be effective, on the one hand, in the treatment of cancer in combination with alkylating agents by suppressing DNA repair and thus driving tumour cells into apoptosis, and on the other hand it appears to be a promising drug target for the treatment of pathologic conditions involving oxidative stress. In view of the existence of several members of the PARP family in mammalian cells, one has to be aware of possible side effects but also of a wide spectrum of potential clinical applications, which calls for the development of more specific inhibitors. © 2004 Wiley‐Liss, Inc.
Molecular detection of circulating tumor cells is an independent prognostic factor in patients with high‐risk cutaneous melanoma
Authors: Mocellin Simone , Del Fiore Paolo , Guarnieri Laura , Scalerta Romano , Foletto Mirto , Chiarion Vanna , Pilati Pierluigi , Nitti Donato , Lise Mario , Rossi Carlo R.
Published: May 2004

Detection of circulating tumor cells (CTCs) might improve current staging procedures by identifying a subgroup of patients with minimal residual disease and thus a higher risk of disease recurrence. Forty patients with ≥2‐mm‐thick cutaneous melanoma with or without lymph node metastasis were enrolled. After standard radical surgery and adjuvant therapy in case of lymph node metastasis, patients were followed up with routine physical and radiologic assessments as well as serial PCR‐based analysis of CTCs using 2 melanoma markers (tyrosinase and Melan‐A/Mart‐1). After a median follow‐up of 30 months, 18 patients had disease recurrence and 28 were PCR‐positive before the disease became clinically evident. The sensitivity of the molecular test was 83%. Median time to PCR positivity and median PCR‐to‐relapse time were 12 and 8 months, respectively. At multivariate analysis, PCR positivity was an independent predictor of disease recurrence (hazard ratio = 2.06, 95% CI 1.07–3.35; p = 0.03). Among high‐risk melanoma patients, serial PCR‐based analysis of CTCs can identify a subgroup at higher risk of disease recurrence, with clinically significant advance. Therefore, CTC detection might be employed for the selection of patients for adjuvant treatment and during follow‐up for early indication of therapeutic failure. © 2004 Wiley‐Liss, Inc.
Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO4in vivo footprinting on its promoter region
Authors: Haga Kazunori , Fujita Hisakazu , Nomoto Minoru , Sazawa Ataru , Nakagawa Koji , Harabayashi Toru , Shinohara Nobuo , Takimoto Masato , Nonomura Katsuya , Kuzumaki Noboru
Published: May 2004

We previously reported that gelsolin gene expression is reduced in various tumors. In an effort to gain further insights into the mechanism of gelsolin downregulation in tumors, we examined the in vivo properties of the gelsolin promoter in urinary bladder cancer cell lines. Neither mutation nor hypermethylation was responsible for gene silencing at the promoter. After exposure to trichostatin A (TSA), a histone deacetylase inhibitor, gelsolin promoter activity was markedly enhanced in the cancer cells, not in cells derived from normal tissue. Chromatin immunoprecipitation assays revealed that both histones H3 and H4 were hypoacetylated in the promoter region of the cancer cells, and the accumulation of acetylated histones was detected by TSA treatment. In vivo footprinting analysis revealed the presence of dimethylsulfate (DMS) hypersensitive site in the untranslated region around nucleotide −35 only in the cancer cells but not in cells derived from normal tissue, and analysis of KMnO4 reactive nucleotides showed that the stem loop structure could be formed in vivo of the cancer cells. This novel stem loop structure may play a part in regulating the transcription of the gelsolin gene in the cancer cells. These results suggest that nucleosome accessibility through histone deacetylation and structural changes (DMS hypersensitivity and stem loop structure) in the promoter region form the basis of the mechanism leading to the silencing of gelsolin gene in human bladder cancer. © 2004 Wiley‐Liss, Inc.