bluuebunny/crossref_metadata_2025 huggingface dataset:
Characteristics of EBV‐infected cells in HIV‐related lymphadenopathy: Implications for the pathogenesis of EBV‐associated and EBV‐unrelated lymphomas of HIV‐seropositive individuals
Authors: Dolcetti Riccardo , Gloghini Annunziata , De Vita Salvatore , Vaccher Emanuela , De Re Valli , Tirelli Umberto , Carbone Antonino , Boiocchi Mauro
Published: February 2007

The present study was performed with the aim of better defining the possible role of Epstein‐Barr‐virus(EBV)‐infected cells in the pathogenesis of HIV‐related lymphadenopathy syndrome (LAS). In addition, since LAS has been considered as a pre‐lymphomatous lesion, we also wished to elucidate the possible contribution of EBV‐carrying cells present in LAS tissues to the development of HIV‐associated malignant lymphomas. To this end, we have characterized EBV‐infected cells in LAS lymph nodes in terms of EBV DNA prevalence, tissue distribution in relation to HlV‐carrying cells, virus sub‐type, expression of latent and replicative antigens, and presence of clonal EBV episomes. When compared with HIV‐unrelated lymphadenopathies (4/10, 40%), LAS showed a higher prevalence of EBV DNA (14/20, 70%). Comparable values of EBV prevalence were detected in LAS with follicular hyperplasia (12/16, 75%) and with follicular involution (4/4, 100%). All EBV+ non‐neoplastic lymph nodes from HIV‐seronegative patients carried type‐1 EBV, whereas LAS specimens showed almost equivalent distribution of the 2 EBV sub‐types. Of the 14 EBV‐carrying LAS, 4 (29%) were positive by Southern‐blot analysis for the BamHI‐W region of the virus genome but negative for the presence of monoclonal EBV episomes. In situ hybridization revealed a remarkably higher load of EBV‐infected cells in LAS than in HIV‐unrelated lymphadenopathies. In LAS lymph nodes, EBV‐carrying cells were identified as isolated, cytologically normal elements, sometimes with immunoblastic morphology, usually scattered throughout the interfollicular areas. By contrast, the expression of HIV p24 was restricted to germinal center cells. All the EBV+ LAS samples were negative for the expression of EBV‐encoded latent (LMP‐1 and EBNA‐2) and replicative proteins (BZLF‐1, BHLF‐1, EA‐D, EA‐R and VCA). In addition, amplification of the immunoglobulin heavy‐chain genes using 2 different polymerase‐chain‐reaction protocols showed evidence of B‐cell clonal expansion in 2/20 (10%) LAS, one EBV− case, and one sample with low numbers of EBV‐infected cells. These results suggest that: (i) EBV‐carrying cells are probably not involved in the development of LAS, either directly or indirectly; (ii) type‐2‐EBV‐infected cells are present in LAS lymph nodes from the early phases of HIV infection; (iii) EBV‐carrying LAS per se probably does not constitute a lesion at high risk for subsequent development of EBV+ lymphomas; (iv) it is unlikely that a high viral load or strong EBV‐mediated antigenic stimulation plays a contributory role in the development of EBV‐unrelated lymphomas of HIV‐seropositive individuals. © 1995 Wiley‐Liss, Inc.
Rapid increase in plasma tenascin‐C concentration after isolated limb perfusion with high‐dose tumor necrosis factor (TNF), interferon gamma (IFNγ) and melphalan for regionally advanced tumors
Authors: Schenk Susanne , Liénard Danielle , Gérain Jean , Baumgartner Murielle , Lejeune Ferdy J. , Chiquet‐Ehrismann Ruth , Rüegg Curzio
Published: February 2007

The matrix protein tenascin‐C (TN‐C) is present in the blood of healthy individuals at concentrations around 1 mg/1. Elevated serum levels have been reported in cancer patients. In this study we have measured the concentration of circulating TN‐C in 40 patients with melanoma, soft‐tissue sarcoma (STS) or squamous‐cell carcinoma (SCC) of the limbs, and have found a minor increase in the mean concentration compared with healthy subjects. Only 10 patients had TN‐C levels above the normal range. No correlation was observed between TN‐C levels and tumor burden. Nineteen patients were treated by isolation limb perfusion (ILP) with TNF, IFNγ, melphalan (11 melanoma, 2 SCC and 1 STS), melphalan alone (3 melanoma) or hyperthermia at 41.5°C (2 melanoma). ILP with TNF, IFNγ and melphalan induced a rapid increase in plasma TN‐C levels, peaking in most patients between 24 to 48 hr after ILP. Two patients treated with hyperthermia only had a slow increase in TN‐C concentration peaking at day 4, while the patients treated with melphalan alone had no significant change. In some cases elevated TN‐C levels persisted for over 8 weeks after ILP. The early rise in TN‐C concentration correlates with the increase in circulating C‐reactive protein. Our findings suggest that circulating TN‐C behaves, at least in part, as an acute‐phase protein and that it may play a role in the inflammatory response. © 1995 Wiley‐Liss, Inc.
Adenovirus‐mediated gene transfer of wild‐type p53 results in melanoma cell apoptosis in vitro and in vivo
Authors: Cirielli Corrado , Riccioni Teresa , Yang Chunlin , Pili Roberto , Gloe Torsten , Chang Joan , Inyaku Kyosuke , Passaniti Antonino , Capogrossi Maurizio C.
Published: February 2007

Gene transfer techniques may provide efficient treatment for a variety of malignant neoplasms. A replication‐deficient adeno‐virus (Ad) vector which carries the cDNA for wild‐type p53 (AdCMV.p53) was tested for its in vitro and in vivo effects on the growth of murine melanoma cell line B16‐G3.26 and human melanoma cell line SK‐MEL‐24. The growth of B16‐G3.26 cells infected with AdCMV.p53 was inhibited when compared to the uninfected cells or cells infected with the control vector AdCMV.NLSβgal. Similarly, the growth of SK‐MEL‐24 cells infected with AdCMV.p53 was also below that of AdCMV.NLSβgal‐infected and uninfected controls. DNA laddering using agarose gel electrophoresis and in situ labeling of DNA fragmentation (TUNEL) showed that AdCMV.p53‐infected murine and human melanoma cells underwent apoptosis. Nude mice injected s.c. either with B16‐G3.26 cells or with SK‐MEL‐24 cells developed localized tumors. These tumors were subsequently infiltrated with either AdCMV.p53, AdCMV.NLSβgal or saline alone. One week after infection, B16‐G3.26 tumors exposed to AdCMV.p53 were 2.5 times smaller than control tumors and exhibited DNA fragmentation. A similar growth‐inhibitory effect of AdCMV.p53 was observed with SK‐MEL‐24 tumors. Thus, Ad‐mediated wild‐type p53 overexpression resulted in melanoma cell apoptosis and inhibition of melanoma growth in vitro and in vivo. These gene therapy approaches may be useful in targeting rapidly growing, malignant melanomas in a clinical setting. © 1995 Wiley‐Liss, Inc.
Enhanced interaction between tubulin and microtubule‐associated protein 2 via inhibition of MAP kinase and CDC2 kinase by paclitaxel
Authors: Nishio Kazuto , Arioka Hitoshi , Ishida Tomoyuki , Fukumoto Hisao , Kurokawa Hirokazu , Sata Makoto , Ohata Masahiro , Saijo Nagahiro
Published: February 2007

Paclitaxel, an anti‐mitotic anti‐cancer agent, is active against solid tumors. The inhibition of depolymerization and promotion of microtubular assembly are essential for the anti‐tumor activity of paclitaxel. Microtubule‐associated proteins (MAPs) co‐polymerize with tubulin and play some roles in microtubular dynamics. We examined the effect of paclitaxel on the interaction between tubulin and MAPs. Human lung‐cancer cells, PC‐14, were synchronized to G1/S border by the thymidine‐double‐block technique. After release from exposure to thymidine, the cells were treated briefly with 2 nM paclitaxel and the levels of α and β tubulins and MAPs were examined after various times. Immunoblot analysis of paclitaxel‐treated cells showed no changes in the overall expression of α and β tubulins, microtubule‐associated protein 2 (MAP2) or MAPs in comparison with controls. The samples were immunoprecipitated with anti‐α‐ and anti‐β‐tubulin antibodies and reblotted with an anti‐MAP2 antibody, which showed that the amount of co‐immuno‐precipitated MAP2 in the synchronized cells, were increased by the brief paclitaxel treatment. These results suggest that paclitaxel treatment enhances the interaction between α and β tubulins and MAP2. Since the phosphorylation state of MAP2 regulates the affinity of MAP2 for tubulins, and mitogen‐activated protein (MAP) kinase is considered to be one of the kinases responsible for MAP2 phosphorylation, the effect of paclitaxel treatment on the MAP‐kinase activity of synchronized PC‐14 cells was examined. Two bands with molecular masses of 42 and 44 kDa were detected by an “intra‐gel” MAP‐kinase assay using myelin basic protein as the substrate. Paclitaxel treatment inhibited the MAP‐kinase activity of PC‐14 cells and inhibition was maximal at the G2/M phase of the cell cycle. Similar, concentration‐dependent inhibition by paclitaxel of cellular MAP kinase of human synchronized small‐cell lung carcinoma, H69, was observed. No inhibition of the MAP kinase of the paclitaxel‐resistant sub‐line H69/T×1 by paclitaxel was observed, suggesting that some change of the MAP‐kinase cascade had occurred in these cells. No direct inhibition of MAP‐kinase activity by paclitaxel was observed in the cell‐free assay (in vitro), suggesting that paclitaxel did not inhibit MAP kinase directly. Since it has been speculated that p34cdc2 kinase is also a kinase that phosphorylates MAP2, the effect of paclitaxel treatment on the p34cdc:2‐kinase activity of synchronized PC‐14 and PC‐9 cells was examined. Paclitaxel inhibited p34cdc2‐kinase activation at the G2/M phase. These results suggest that paclitaxel inhibited MAP kinase and p34cdc2 kinase in vivo indirectly. These actions of paclitaxel may be responsible for the increased affinity between MAP2 and tubulins that it induces. © 1995 Wiley‐Liss, Inc.
Fibroblast growth factor 3 is tumorigenic for mouse mammary cells orthotopically implanted in nude mice
Authors: Hajitou Amin , Calberg‐Bacq Claire‐ M.
Published: February 2007

Fibroblast growth factor‐3 (Fgf‐3) is involved in mouse mammary tumorigenesis since the Fgf‐3 gene is a main target for mouse mammary tumor virus (MMTV) insertional activation. Its action has been correlated with the appearance of pregnancy‐dependent tumors. We describe here the effects on normal mouse mammary EF43 cells of the short Fgf‐3 protein form which enters the secretory pathway. The genes, Fgf‐3 AUG or Fgf‐4 for comparison, were introduced in the mammary cells by means of retroviral vectors. Fgf‐3 expression did not modify EF43 cell morphology, had no effect on growth in soft agar nor on the inhibitory action exerted on cell growth by TGF‐β; however, it allowed the cells to grow under low serum conditions in the absence of insulin and EGF. The Fgf‐3‐expressing cells were not tumorigenic in nude mice when injected s.c., but tumors developed when the cells were implanted in the mammary gland. The tumors appeared after some latency; they had a slow growth phase followed by a phase of increased growth rate. An identical tumoral growth pattern was observed in ovariectomized nude mice. These results show that the secreted Fgf‐3 form can initiate tumorigenesis and that the induced tumors are hormone‐independent. The mammary‐gland environment, however, is required for the EF43 cells to grow and differentiate. During that process, which resembles natural cell growth during mammary‐gland development at pregnancy, the cells could pass through a stage which is specifically sensitive to Fgf‐3. © 1995 Wiley‐Liss, Inc.
Expression of estrogen, progesterone and epidermal growth factor receptors in primary and metastatic breast cancer
Authors: van Agthoven Ton , Timmermans Mieke , Dorssers Lambert C. J. , Henzen‐Logmans Sonja C.
Published: February 2007

The prognostic value of epidermal growth factor receptor (EGFR) expression and its biological role in estrogen receptor‐positive (ER+) and ER‐negative (ER−) primary breast cancer is controversial. In this study, distributions of ER, progesterone receptor and EGFR have been established using immunohisto‐chemistry in both primary breast tumors and their matched axillary lymph node metastases of 26 patients or their matched distant metastases of 2 patients. In addition, 5 patients with bilateral breast cancer were studied. ER+ tumor cells were detected in 22 (69%) and EGFR+ tumor cells were detected in 11 (34%) primary breast carcinomas. Expression of ER and EGFR was inverse regarding the individual tumor cells in both primary tumors and metastases. Relationship of EGFR expression with poorly differentiated and large breast tumors was observed. Furthermore, primary tumors with a predominant lobular component were ER+ and, with one exception, EGFR−. Invasive ductal carcinomas were more frequently EGFR+. No apparent differences in receptor expression were observed between primary tumors and lymph node metastases or chro‐nously or metachronously occurring bilateral breast cancers. Only one ER+ primary tumor showed a switch to EGFR expression in the involved lymph node. Our study shows that a shift in receptor phenotype between primary tumors and lymph node metastases is a rare event and, thus, additional analyses of involved lymph nodes will not likely serve as a better predictor for response to anti‐estrogen therapy. We conclude that expression of EGFR is not a prerequisite for development of metastases. © 1995 Wiley‐Liss, Inc.
High frequency of K‐ras mutations in normal appearing lung tissues and sputum of patients with lung cancer
Authors: Yakubovskaya Marina S. , Spiegelman Vladimir , Luo Feng C. , Malaev Serge , Salnev Alexander , Zborovskaya Irina , Gasparyan Alexander , Polotsky Boris , Machaladze Zurab , Trachtenberg Alexander C. , Belitsky Gennady A. , Ronai Zeev
Published: February 2007

To evaluate the possible use of mutant ras as a biomarker for lung cancer, we have analyzed “normal appearing” lung tissue, lung tumor, lung metastases and sputum samples from patients with non‐small cell lung cancer (NSCLC). As a control, we used lung tissue and sputum samples from patients without oncological diseases or lung disorders. Our analyses were performed with the aid of enriched PCR (EPCR), a method which enables detection of ras mutation even if present at low incidence. EPCR identified K‐ras codon 12 mutations in 10% of lung tissues obtained from patients with no lung diseases, whereas the same mutation was detected in 60% of samples of normal appearing lung tissues obtained from patients with NSCLC, 62% of NSCLC tumors and 80% of metastases. Analysis of sputum samples of patients with NSCLC identified 47% to harbor mutant ras allele, whereas 12.5% of controls diagnosed with non‐oncological lung diseases carried this mutation. Most of these mutations were detected with the aid of EPCR only, indicating that a minority of cells in a given sample harbor this mutation. The ability to detect K‐ras codon 12 mutation in 60% of lung tissue samples and in 47% of sputum samples taken from patients with lung cancer (as compared with 10% and 12.5% of respective controls) points to the potential use of ras mutation as a biomarker for exposure and possible identification of patients who may be at a higher risk of developing lung cancer. © 1995 Wiley‐Liss, Inc.
Invasive phenotype of MCF10A cells overexpressing c‐Ha‐ras and c‐erbB‐2 oncogenes
Authors: Giunciuglio Daniela , Culty Martine , Fassina Gianfranco , Masiello Luciana , Melchiori Antonella , Paglialunga Giuseppina , Arand Gloria , Ciardiello Fortunato , Basolo Fulvio , Thompson Erik W. , Albini Adriana
Published: February 2007

Infection with erbB‐2 (E) of Ha‐roj (H) oncogene‐transfected cells has been previously shown to cooperatively induce anchorage‐independent growth of the MCF10A human mammary epithelial cell line in vitro, but not to induce nude mouse tumorigenicity. Here we show that oncogene‐transformed MCF10A are able to halt in the lungs of nude mice, a sign of organ colonization potential. We have therefore studied the transformants for in vitro migratory and invasive properties known to correlate with the metastatic potential of human mammary carcinoma cells in nude mice. MCF10A transfected with Ha‐ras, infected with a recombinant retroviral vector containing the human c‐erbB‐2 proto‐oncogene (MCF10A‐HE cells), show a higher invasive index than either the single transfectant (MCF10A‐H) or MCF10A‐erbB‐2 (MCF10A‐E) cells in the Boyden chamber chemotaxis and chemoinvasion assays. The MCF10A‐HE cells also adopted an invasive stellate growth pattern when plated or embedded in Matrigel, in contrast to the spherical colonies formed by the single transformants MCF10A‐H, MCF10A‐E, and the parental cells. Dot‐blot analysis of gelatinase A and TIMP‐2 mRNA levels revealed increasing gelatinase A mRNA levels (HE > E > H > MCF10A) and reduced TIMP‐2 expression in both single and double transformants. Furthermore, MCF10A‐HE cells show more MMP‐2 activity than parental MCF 10A cells or the single transformants. CD44 analysis revealed differential isoform banding for the MCF10A‐HE cells compared to parental ceils, MCF10A‐H and MCF10A‐E, accompanied by increased binding of hyaluronan by the double transformants. Our results indicate that erbB‐2 and Ha‐ros co‐expression can induce a more aggressive phenotype in vitro, representative of the malignancy of mammary carcinomas. © 1995 Wiley‐Liss, Inc.
Anti‐tumor activity of tumor necrosis factor in combination with interferon‐γ is not affected by prior tolerization
Authors: Takahashi Nozomi , Fiers Walter , Brouckaert Peter
Published: February 2007

Repetitive administration of low doses of tumor necrosis factor (TNF) results in a state of selective tolerance to some of its effects. We have demonstrated that tolerance does not impair the therapeutic efficacy of TNF against a syngeneic murine B16BL6 melanoma and allows a complete cure. Another study, performed with a distinct tumor model, came to apparently contradictory results. To clarify this, we investigated whether the outcome depended on the tumor type and on the inclusion of interferon‐γ (IFNγ) in the treatment. Three syngeneic tumors of different histological origin, i.e., B16BL6 melanoma, Lewis lung carcinoma (LLC) and EL4 lymphoma, were compared in C57BL/6 mice. The anti‐tumor efficacy of TNF against BI6BL6 and EL4 was not impaired in tolerant mice, but the effect of TNF against LLC was slightly, though significantly, reduced. Inclusion of IFNγ in the treatment regimen, however, abolished this difference and resulted in complete cure for all 3 tumor systems. As therapeutically optimal doses were lethal in normal mice, only tolerance allowed a long‐term cure. We conclude that the influence of tolerance on the anti‐tumor activity of TNF as a single agent depends on the tumor type; in combination therapy with IFNγ, however, tolerance allowed us to dissociate lethal toxicity from anti‐tumor activity, irrespective of the tumor type tested. © 1995 Wiley‐Liss, Inc.
Prostate‐specific antigen activates single‐chain urokinase‐type plasminogen activator
Authors: Yoshida Etsuo , Ohmura Sayuri , Sugiki Masahiko , Maruyama Masugi , Mihara Hisashi
Published: February 2007

Prostate‐specific antigen (PSA) increases in the plasma of patients with prostate cancer, and has therefore been used as a reliable tumor marker. It has been demonstrated that prostate cancer cells over‐express urokinase‐type plasminogen activator (uPA), which plays an important role in tumor invasion and metastasis. We found that PSA converts the single‐chain pro‐form of urokinase‐type plasminogen activator (scuPA) to an active 2–chain form. The active 2–chain uPA generated from scuPA by PSA was measured by hydrolyzation of S‐2444, a synthetic substrate for uPA. PSA activated scuPA time‐ and dose‐dependently. SDS‐PAGE analysis revealed that, after incubation with PSA, the intensity of the 55–kDa band of scuPA decreased concomitantly with increases in the intensity of the 2 bands at 33 kDa and 22 kDa. Amino‐acid‐sequence analysis indicated that PSA cleaved Lys158‐Ile159, which corresponds with the site cleaved by plasmin. PSA did not enhance or impair the activity of the 2–chain form of uPA. These findings imply that PSA could be an initiator of the protease cascade involved in prostate‐cancer invasion and metastasis. © 1995 Wiley‐Liss, Inc.
Malignant lymphoma induction in rabbits by intravenous inoculation of Epstein‐Barr‐virus‐related herpesvirus from HTLV‐II‐transformed cynomolgus leukocyte cell line (Si‐IIA)
Authors: Hayashi Kazuhiko , Koirala Tirtha R. , Ino Hideo , Chen Hong‐Li , Ohara Nobuya , Teramoto Norihiro , Yoshino Tadashi , Takahashi Kiyoshi , Yamada Masao , Nil Shiro , Miyamoto Kanji , Fujimoto Koji , Yoshikawa Yasuhiro , Akagi Tadaatsu
Published: February 2007

Maignant lymphomas, which were usually of T‐cell type, were Induced in 10 of 13 (77%) male rabbits (Japanese white, 8/10; New Zealand white, 2/3) inoculated i.v. with HTLV‐II‐transformed simian (Cynomolgus) leukocyte cell line (Si‐IIA) cells. Of 7 rabbits injected with cell‐free pellets from Si‐IIA cultures, 5 also developed malignant lymphoma (15–28 days). Lymphoma development was completely inhibited by inactivation of cell‐free pellets from Si‐IIA culture with ethyl ether and was almost suppressed by neutralization of the cell‐free pellets with anti‐Si‐IIA sera. Herpesvirus particles were discovered very rarely in Si‐IIA cells, in addition to C‐type virus particles, by electron microscopy. Si‐IIA cells were positive for Epstein‐Barr‐virus(EBV)‐associated nuclear antigen (EBNA) by immunofluo‐resconce (IF) test. Antibody response to viral capsid antigen of EBV was also detected in sera from rabbits inoculated with Si‐IIA. EBV‐encoded RNA‐I (EBER‐I) was demonstrated in Si‐IIA, the tumor tissues and all rabbit tumor cell lines by in situ hybridization. EBV DNA was also detected in Si‐IIA and rabbit lymphoma cell lines by polymerase chain reaction (PCR) and Southern blotting. However, EBV DNA was amplified only by some primers complementary to human EBV sequence (B95–8), but not by other primers. Integration of HTLV‐II provirus genome could not be detected in Si‐IIA‐induced rabbit tumor cells. Moreover, no lymphoma was induced by inoculation of HTLV‐IIC and MOT (other HTLV‐II‐producing human cell lines), B95‐8 (EBV‐producing cell line) or TALL‐1 and peripheral leukocytes from normal Cynomolgus (controls). Neither Herpesvirus saimiri nor H. ateles (simian oncogenic viruses) were detected in Si‐IIA cells by IF test. These data suggest that the high rate of lymphoma induction in rabbits may not be caused by HTLV‐II, human EBV (B95–8) or well‐known simian oncogenic viruses, but by EBV‐related herpesvirus derived from Si‐IIA cells or HTLV‐IIA cells, with which Si‐IIA was established. The availability of this animal model promises to clarify the role of EBV in human lymphoma and provides a means of studying prophylactic and therapeutic regimens. © 1995 Wiley‐Liss, Inc.
Familial clustering of breast cancer: Possible interaction between dna repair proficiency and radiation exposure in the development of breast cancer
Authors: Helzlsouer Kathy J. , Harris Emily L. , Parshad R. , Fogel Sandy , Bigbee William L. , Sanford Katherine K.
Published: February 2007

The capacity of peripheral blood lymphocytes to repair X‐ray‐induced DNA damage, manifest as chromatid damage 30‐90 min after G2‐phase X‐irradiation, was measured among available members of a family exhibiting a cluster of breast‐cancer cases occurring in one generation. The cancer patients had been exposed to repeated chest fluoroscopic examinations during early childhood and adolescence. The development of breast cancer was correlated with DNA repair proficiency and history of radiation exposure. The results of the family study provide preliminary support for the hypothesis that a deficiency in repair of X‐irradiation DNA damage may be a susceptibility factor for the development of breast cancer. This hypothesis, however, requires confirmation in a larger study. Studying the combined effect of susceptibility factors and environmental exposures may enhance our knowledge of the etiology of breast cancer and provide leads for effective prevention strategies aimed at reducing exposures or altering susceptibility to unavoidable exposures. © 1995 Wiley‐Liss, Inc.
Expression of mage genes in transitional‐cell carcinomas of the urinary bladder
Authors: Patard Jean‐Jacques , Brasseur Francis , Gil‐Diez Sixtina , Radvanyi François , Marchand Marie , François Philippe , Abi‐Aad Antoine , Van Cangh Paul , Abbou Clément Claude , Chopin Dominique , Boon Thierry
Published: February 2007

Human genes MAGE‐1 and MAGE‐3 code for distinct antigens, which are recognized on melanoma cells by autologous cytolytic T lymphocytes (CTL). These antigens may constitute useful targets for anti‐cancer immunotherapy, since no expression of MAGE genes has been observed in normal tissues other than testis. Out of 57 samples of primary transitional‐cell carcinomas of the bladder, 12 (21%) expressed MAGE‐1 and 20 (35%) expressed MAGE‐3. All but one of the tumors expressing MAGE‐1 also expressed MAGE‐3. Genes MAGE‐2 and MAGE‐4, which are closely related to MAGE‐1 and MAGE‐3, were expressed by 30% and 33% of the tumors respectively. MAGE expression was more frequent in advanced tumor stages: 61% of the invasive tumors (stage ± T2) were positive for expression of at least one of the four genes, whereas only 28% of the superficial tumors (stages Ta and Tl) expressed these genes. © 1995 Wiley‐Liss, Inc.
Expression of platelet‐derived endothelial cell growth factor/thymidine phosphorylase in human breast cancer
Authors: Toi Masakazu , Hosnina Seigo , Taniguchi Tadaaki , Yamamoto Yutaka , Ishitsuka Hideo , Tominaga Takeshi
Published: February 2007

We have investigated the expression of platelet‐derived endothelial‐cell growth factor/thymidine phosphorylase (PD‐ECGF/dThdPase) in human breast‐cancer tissues by the immunocytochemical method using anti‐PD‐ECGF/dThdPase monoclonal antibody. Out of 100 invasive‐ductal‐carcinoma tissue samples, 39 (39%) were evaluated as PD‐ECGF/dThdPase‐positive. The expression of PD‐ECGF/dThdPase was identified mainly in the cytoplasma of tumor cells. The expression of PD‐ECGF/dThdPase was significantly associated with the microvessel density assessed by immunostaining to factor‐VIII‐related‐antigen (p < 0.05). However, there was no correlation between expression of PD‐ECGF/dThdPase and menopausal status, tumor size, axillary lymph‐node metastases, hormone‐receptor status, epidermal‐growth‐factor receptor, or erb‐B‐2‐protein and p53‐protein expression. We suggest that expression of PD‐ECGF/dThdPase plays an important role in the promotion of angiogenesis in human breast cancer. © 1995 Wiley‐Liss, Inc.
Comparative analysis of p53 gene mutations and protein accumulation in human non‐small‐cell lung cancer
Authors: Top Bert , Mooi Wolter J. , Klaver Siegina G. , Boerrigter Lucie , Wisman Peter , Elbers Hans R. J. , Visser Sjoerd , Rodenhuis Sjoerd
Published: February 2007

A series of 54 resected primary non‐small‐cell lung carcinomas was analyzed for p53 gene mutations and for p53 protein accumulation and the findings were correlated with clinical parameters. Mutations in exons 5 through 8 of the p53 gene were identified by a denaturing gradient gel electrophoresis (DGGE) assay and cycle sequencing, whereas p53 protein accumulation was detected in paraffin‐embedded tissue by immunostaining using 2 different murine monoclonal antibodies (MAbs) (BP53‐12 and DO7). A p53 gene mutation and/or p53 protein accumulation was found in 37 of 54 tumors. Mis‐sense mutations were closely associated with positive immunostaining, which was intense in 15 out of 17 cases with a mutation. In 10 tumors, obvious p53 accumulation was detected in the absence of mutations in exons 5 through 8. Conversely, only one of 8 p53 non‐sense mutations led to detectable p53 accumulation. The most frequent single base changes were G → T transversions and C → T transitions. The presence of a p53 alteration was not related to age, tumor size, stage or histology. However, we found a significant inverse correlation between p53 alterations and the presence of a K‐ras mutation. This was reflected in the overall postoperative survival data: patients with p53 alterations in their tumors tended to have a better prognosis than those without a p53 alteration; however, this difference was lost when cases with a K‐ras mutation were omitted from the analysis. © 1995 Wiley‐Liss, Inc.
Chromosome‐9 loss detected by fluorescence in situ hybridization in bladder cancer
Authors: Sauter Guido , Moch Holger , Carroll Peter , Kerschmann Russel , Mihatsch Michael J. , Waldman Frederic M.
Published: February 2007

A loss of chromosome‐9 material is one of the most frequent genomic aberrations known in bladder cancer. In order to better understand the role of chromosome‐9 losses in bladder cancer, 125 formalin‐fixed and 37 unfixed bladder tumors were examined using fluorescence in situ hybridization (FISH). A repetitive probe for a pericentromeric region on 9q12 (pHUR98) was applied for chromosome‐9 copy‐number enumeration. Under‐representation of chromosome 9 was found in 74 of 162 cases. There was no association between loss of chromosome 9 and increased grade or stage, papillary growth pattern, p53 protein expression, or tumor‐cell proliferation (Ki‐67). These data show that chromosome‐9 loss is an early event in bladder‐cancer development, occuring independently of p53 alterations. In order to determine the prevalence of large sub‐regional chromosome‐9 deletions, dual hybridizations with pHUR 98 and cosmid probes for 9q34, 9q22, and 9p21 were performed. Partial deletion was detected in only 1 of 36 cases for 9q34 and in 1 of 24 cases for 9p21. Surprisingly, amplification of the interferon alpha locus on 9p21 was seen in 1 of 24 tumors. The finding of 9p amplification may indicate the site of an oncogene relevant for bladder cancer. © 1995 Wiley‐Liss, Inc.
High frequency of allelic imbalance at chromosome region 16q22‐23 in human breast cancer: Correlation with high pgr and low s phase
Authors: Skirnisdottir Sigurlaug , Eiriksdottir Gudny , Baldursson Trausti , Barkardottir Rosa B. , Egilsson Valgardur , Ingvarsson Sigurdur
Published: February 2007

The loss of genetic material from a specific chromosome region in tumors suggests the presence of tumor‐suppressor genes. Loss of heterozygosity (LOH) or allelic imbalance (AI) on the long arm of chromosome 16 is a known event in sporadic breast cancer. To locate the commonly deleted regions, and therefore (a) candidate tumor‐suppressor gene(s), a deletion map of chromosome 16 was made, using 10 microsatellite markers on 150 sporadic breast tumors. The 3 smallest regions of overlap (SRO) were detected on the long arm of chromosome 16. Allelic imbalance was observed with at least one marker in 67% of the tumors. One marker, D16S421, at the 16q22‐23 region, showed the highest allelic imbalance, 58%. Tumors with and without AI on 16q were tested for correlation with clinico‐pathological features of the tumors such as estrogen‐ and progesterone‐receptor content (ER and PgR), age at diagnosis, tumor size, node status, histological type, S‐phase fraction, AI on chromosome 3p, and ploidy. A correlation was found between AI on 16q and high PgR content, also low S‐phase fraction (99% confidence limits). A comparison of tumors with and without AI at the D16S421 marker locus revealed a slight correlation with high PgR content. The survival data showed no difference between patients with AI on 16q and those with a normal allele pattern on the long arm of chromosome 16. © 1995 Wiley‐Liss, Inc.
ras, p53 and hpv status in benign and malignant prostate tumors
Authors: Moyret‐Lalle Caroline , Marcais Christophe , Jacquemier Jocelyne , Moles Jean‐Pierre , Daver Alain , Soret Jean‐Yves , Jeanteur Philippe , Ozturk Mehmet , Theillet Charles
Published: February 2007

To study the role of ras, p53 genes and HPV virus (16 and 18) in the development of prostate cancer, we analyzed tissue sections from 27 patients affected with carcinomas (stages A to D) and from 24 patients with adenomas. Mutations of H, K and N‐ras and p53 (exons 2‐9) were studied by SSCP and DNA sequencing. Accumulation of p53 protein was studied by immu‐nohistochemistry on tissue sections. Tumors were also analyzed for the presence of HPV 16 and ‐18 sequences by PCR and DNA hybridization with sequence‐specific oligonucleotides. No mutation was found in the three ras genes studied, either in carcinomas or adenomas. By SSCP analysis we identified p53 mutations in only 2 of 19 carcinomas studied, both in exon 7. Immunohistochemical results strongly correlate with the SSCP results: p53 protein was positive in tumors with p53 mutation but not in others; 32% of studied adenomas had detectable HPV16 DNA, while 53% of carcinomas were HPV16+. Among these I presented a p53 mutation. No HPV 18 E6 sequence could be detected. Our data show that in prostate tumors from France, mutations of p53 and ras are rare events but that these tumors display detectable HPV 16 DNA at a high frequency. The low incidence of p53 mutation, associated to a significant proportion of tumors showing HPV16 DNA, could suggest that in prostate cancer HPV 16 infection could participate in p53 inactivation by E6. © 1995 Wiley‐Liss, Inc.
Detection of mage‐4 protein in lung cancers
Authors: Shichijo Shigeki , Hayashi Akihiro , Takamori Shinzo , Tsunosue Rika , Hoshino Tomoaki , Sakata Motoko , Kuramoto Terukazu , Oizumi Kotaro , Itoh Kyogo
Published: February 2007

Expression of genes of the MAGE family, which encode tumor‐rejection antigens recognized on HLA‐AI and ‐Cw1601 by cytotoxic T lymphocytes (CTL), was investigated in lung cancers at the mRNA [MAGE‐1, ‐2, ‐3/‐6, and ‐4 (4a and/or 4b)] and protein (MAGE‐4) levels. MAGE‐1, ‐2, ‐3/‐6 and ‐4 genes were expressed, respectively, at the mRNA level in 6, 7, 20 and 7 of 53 lung cancers (50 non‐small‐cell lung cancers and 3 small‐cell lung cancers) by the reverse transcription‐polymer‐ase chain reaction (RT‐PCR) method. Polyclonal antibody (Ab) and monoclonal antibody (MAb) against recombinant MAGE‐4b protein were developed to detect MAGE‐4 protein. Both the polyclonal Ab and the R5 MAb recognized a 45‐kDa protein in extracts of MAGE‐4 mRNA‐positive lung cancers, and showed no apparent cross‐reactivity with the other MAGE gene products except with MAGE‐4a by immunoblot analyses and transfec‐tion experiments. MAGE‐4 protein was detected on 13 of 44 (30%) lung cancers (18 to 55,989 pg/mg) by ELISA with the polyclonal Ab and R5 MAb. These 13 lung cancers consisted of 6 of 6 MAGE‐4 mRNA‐detectable and 7 of 38 MAGE‐4 MRNA‐undetectable lung cancers. Histologically, these comprised 7 of 10 squamous‐cell carcinomas, 4 of 30 adenocarcinomas and 2 of 3 small‐cell lung cancers. The proportions of MAGE gene‐positive samples, at both the mRNA and protein levels, correlated with the size of the primary tumors and with regional node involvement. These results should provide important information on specific immunotherapy of lung cancers using MAGE gene products. © 1995 Wiley‐Liss, Inc.
Predictive value of statin, a g‐associated cell cycle protein, in childhood acute lymphoblastic leukemia
Authors: Volmu Manfred , Hecker Stefanie , Sauerbrey Axel , Mattern Jurgen
Published: February 2007

Blast cells of 71 children with newly diagnosed acute lympho‐blastic leukemia (ALL) were examined with a novel monoclonal antibody (MAb), S‐44, immunoreactive to statin, a nuclear protein specifically expressed in non‐proliferating cells. The statin labeling index (LI) varied greatly from case to case and ranged from 0.3% to 86%, with a mean value of 10%. The degree of statin LI correlated inversely with the expression of Ki‐67. When patients were subdivided according to whether the statin LI was higher or lower than the mean value, patients with higher statin LI did not respond to chemotherapy and survived for a shorter period of time. Our results suggest that labeling of statin with the S‐44 MAb is a useful prognostic marker of the cytotoxic effects of anti‐cancer drugs in patients with ALL. © 1995 Wiley‐Liss, Inc.
c‐erbb growth‐factor‐receptor proteins in ovarian tumours
Authors: Simpson Barbara J. B. , Phillips Hamish A. , Lessells Alastair M. , Langdon Simon P. , Miller William R.
Published: February 2007

Immunohistochemical expression of EGF‐R, c‐erbB‐2 and c‐erbB‐3, members of the type‐l family of receptor tyrosine kinases, were investigated in 67 primary ovarian‐tumour samples (46 malignant, 8 borderline and 13 benign), and related to tumour clinicopathological features. The incidence of all 3 receptor proteins was highest in overtly malignant tumours. No significant correlations were observed between either EGF‐R or c‐erbB‐3 and clinical parameters such as tumour stage, differentiation or extent of debulking surgery, but c‐erbB‐2 was significantly associated with several indicators of prognosis, including early stage and good/moderate differentiation in optimally debulked tumours. Multiple expression of c‐erbB receptor proteins was also significantly higher in malignant tumours compared with borderline and benign tumours. Early‐stage tumours were also more likely to express multiple c‐erbB‐receptor proteins than were late‐stage tumours. Co‐expression of EGF‐R with c‐erbB‐2, and c‐erbB‐2 with c‐erbB‐3 was significantly greater in malignant tumours than in borderline or benign tumours, and within the malignant tumour group, positive associations were observed between EGF‐R and c‐erbB‐3, also between c‐erbB‐2 and c‐erbB‐3. Because of the evidence of increased expression of individual c‐erbB proteins as well as multiple expression of this family of growth‐factor receptors in malignant ovarian tumours, we hypothesize that stimulation by the appropriate ligands may confer a selective advantage to cells expressing more than one receptor. Increased expression of c‐erbB growth‐factor receptors in malignancy may mediate increased propensity for tumour development. © 1995 Wiley‐Liss, Inc.
Expression of the mage gene family in primary and metastatic human breast cancer: Implications for tumor antigen‐specific immunotherapy
Authors: Russo Vincenzo , Traversari Catia , Verrecchia Alessandro , Mottolese Marcella , Natali Pier Giorgio , Bordignon Claudio
Published: February 2007

Some human tumors express known antigens that can be utilized as targets for specific immunotherapy. An absolute requirement for the efficacy of this therapeutic strategy is an adequate expression of the candidate antigen by all cells of the primary and metastatic tumor. To examine the presence and distribution of tumor‐associated antigens in metastatic breast cancer, we used PCR analysis and ethidium bromide staining to test the expression of genes of the MAGE family in 28 primary tumors and related metastatic samples. Overall, samples obtained from 7 of 28 patients revealed positive. However, 2 of 3 primary tumors positive for MAGE‐1 and/or MAGE‐3 had corresponding negative metastatic lesions. On the contrary, 4 of the 25 MAGE‐negative primary tumors gave rise to positive metastatic nodes. Our results confirm in vivo, at the molecular level, the tumor‐antigen heterogeneity previously observed at the cellular level by in vitro analysis. Our data strongly suggest that, at least in patients with breast cancer, multiple different antigens would be required to optimize the recognition of neoplastic cells in immunotherapeutic protocols using MAGE products as target antigens. © 1995 Wiley‐Liss, Inc.
Immunocytochemical localization of fos protein in human breast cancers and its relationship to a series of prognostic markers and response to endocrine therapy
Authors: Gee Julia M. W. , Ellis Ian O. , Robertson John F. R. , Willsher Peter , McClelland Richard A. , Hewitt Katherine N. , Blamey Roger W. , Nicholson Robert I.
Published: February 2007

The nuclear transcription factor Fos is inducible by both steroid hormones and peptide growth factors. It thus forms a potential point of interaction between steroid hormone‐ and growth factor‐directed pathways and may be critical in the subversion of steroid hormone control in breast cancer. In this light, the present study has used immunocytochemistry to demonstrate in clinical primary breast cancer that Fos expression is indeed significantly associated with a failure to respond to endocrine therapy, with preliminary analysis revealing a survival advantage for those patients whose tumours lacked Fos. Sustained elevated levels of Fos expression were significantly associated with further factors, notably peptide growth factors and their receptors (e.g., EGFR, TGFα), as well as with the proliferation marker Ki‐67, which have been linked previously to endocrine insensitivity in breast cancer. In contrast, there appeared to be a trend for Fos to be absent in those tumours expressing markers of endocrine responsiveness (e.g., oestrogen receptor [ER], and also ER‐mediated markers i.e., PR, pS2 or bcl‐2). Interestingly, many of these trends were maintained in ER+ patients, suggesting that Fos may be of importance in directing loss of endocrine sensitivity in ER+ disease. © 1995 Wiley‐Liss, Inc.
Peptide‐specific ctl in tumor‐infiltrating lymphocytes from metastatic melanomas expressing mart‐1/melan‐a, gp100 and tyrosinase genes: A study in an unselected group of hla‐a2.1‐positive patients
Authors: Spagnoli Giulio C. , Schaefer Christoph , Willimann Thomas E. , Kocher Thomas , Amoroso Antonio , Juretic Antonio , Zuber Markus , Luscher Urs , Harder Felix , Heberer Michael
Published: February 2007

Peptide specificity of cultured tumor‐infiltrating lymphocytes (TIL) was systematically investigated in a group of HLA‐A2.1+ metastatic melanoma patients consecutively referred to our department for surgical treatment. Seven samples from 6 patients were studied. All surgical specimens showed evidence of gp100, MART‐1/Melan‐A and Tyrosinose gene expression as detectable by reverse PCR (rPCR). Cultured TIL from 2 patients displayed cytotoxic activity against autologous or HLA‐matched EBV‐transformed cells previously pulsed with MART‐1/Melan‐A27‐35 peptide. In contrast, no CTL activity against gp 100280‐288 or tyrosinase1‐9 peptides could be observed. TIL were then repeatedly stimulated in vitro with the same peptides. After 6 restimulation courses at weekly intervals, specific recognition of gp100280‐288 and MART‐1/Melan‐A27‐35 peptides was detectable in 3 and 5 TIL populations, respectively. In one case Tyrosinase1‐9‐specific CTL could be demonstrated. Two TIL populations from metastases resected from a melanoma patient at 6 months' distance showed a different peptide specificity pattern, and no specific CTL could be generated from simultaneously sampled peripheral blood mononuclear cells (PBMC). All peptide‐specific CTL populations also displayed significant cytotoxic activity against HLA‐A2.1 matched melanoma cell lines expressing the antigens under investigation. Our data indicate that CTL specific for MART‐Melan‐A27‐35, gp100280‐288 or Tyrosinase1‐9 peptides could be expanded with varying frequency from TIL derived from 4 out of 6 HLA‐A2.1+ patients whose tumors expressed the genes encoding these tumor‐associated antigens (TAA). © 1995 Wiley‐Liss, Inc.
Preferential expression of the multidurg‐resistance‐associated protein (MRP) in adenocarcinoma of the lung
Authors: Sugawara Isamu , Yamada Hiroyuki , Nakamura Hisayoshi , Sumizawa Tomoyuki , Akiyama Shinichi , Masunaga Atsuko , Itoyama Shinji
Published: February 2007

The expression of multidrug‐resistance‐associated protein (MRP) was assessed in various types of untreated lung cancer using an immunohistochemical technique. MRP was abundantly expressed in 28 of 59 adenocarcinoma specimens (47%) and its expression was associated with the degree of glandular differentiation of the tumor. MRP expression in well‐differentiated adenocarcinomas (56%) was higher than in poorly differentiated adenocarcinomas (22%) (p < 0.01).

In contrast, MRP expression in other cancer types was lower—20% in squamous‐cell carcinomas, 20% in large‐cell carcinomas and 0% in small‐cell carcinomas and carcinoids. RT‐PCR showed that the MRP‐positive adenocarcinomas and squamous‐cell carcinomas expressed mrp mRNA significantly. Immunoelectron microscopically, MRP was localized in the plasma membrane and rough endoplasmic reticulum. It is thus important to take MRP into account when considering chemotherapy for lung cancers because levels of mdr 1 gene product, another multidrugresistance gene family, are low in untreated lung cancers. © 1995 Wiley‐Liss, Inc.
MDM2 gene amplification in bone and soft‐tissue tumors: Association with tumor progression in differentiated adipose‐tissue tumors
Authors: Nakayama Tomitaka , Toguchida Junya , Wadayama Bun‐Ichiro , Kanoe Hiroshi , Kotoura Yoshihiko , Sasaki Masao S.
Published: February 2007

We have studied 107 bone and soft‐tissue sarcomas and 8 lipomas for amplification of the MDM2 gene. This gene was amplified in 3 out of 67 osteosarcomas, 3 out of 20 malignant fibrous histiocytomas, 4 out of 20 liposarcomas, and 4 out of 8 lipomas. The amplification was associated with overexpression of mRNA. In osteosarcomas, contrary to previous findings, all amplifications were observed in primary lesions. In liposarcomas, the amplification was seen exclusively in well‐differentiated tumors with high frequency (4/5) but not in other subtypes (0/15). In addition, MDM2 amplification was also frequently found in deep‐seated intra‐ or intermuscular lipomas (4/5). Hence, it is suggested that MDM2 amplification plays a significant role in the development of differentiated adipose‐tissue tumors. Three well‐differentiated liposarcomas with MDM2 amplification coexisted with high‐grade dedifferentiated sarcomas, in which MDM2 amplification was also observed. Interestingly, in 2 of these cases, the grades of amplification correlated with the histological grades, indicating an important role of MDM2 overexpression in tumor progression. © 1995 Wiley‐Liss, Inc.
Incidence of germ‐line p53 mutations in patients with gliomas
Authors: Li You‐Jun , Sanson Marc , Hoang‐Xaun Khê , Delattre Jean‐Yves , Poisson Michel , Thomas Gilles , Hamelin Richard
Published: February 2007

Epidemiological studies on intracranial tumors have suggested that the observed familial aggregation of a proportion of gliomas may be due to inherited predisposition to their development. In the Li‐Fraumeni syndrome (LFS) associated with germ‐line mutations of the p53 gene, nervous‐system tumors are observed with increased frequency. However, the contribution of germ‐line p53 mutation to the incidence of brain tumors has not been investigated. In order to address this point, we have performed 2 independent investigations. First, we have examined an unselected series of brain tumors. Whenever the presence of a p53 mutation in the tumor was observed, the possible germ‐line origin of the mutation was investigated. Germ‐line p53 mutations were also analyzed in constitutional DNA of patients with gliomas that had been selected for an unusual personal or familial history of cancer. Germ‐line p53 mutations were detected in 1 out of 80 unselected cases and in 3 out of 15 selected cases (20%). We conclude that germ‐line p53 mutation may contribute to a small fraction of gliomas that develop in the general population. The presence of a personal or familial history of cancer in a patient with glioma should prompt the search for a germ‐line p53 mutation. However, the low frequency of p53 germ‐line mutation suggests that alterations of this gene may not account for most familial cases of gliomas. © 1995 Wiley‐Liss, Inc.
Risk modifiers in carriers of brca1 mutations
Authors: Narod Steven A. , Goldgar David , Cannon‐Albright Lisa , Weber Barbara , Moslehi Roxanna , Ives Elizabeth , Lenoir Gilbert , Lynch Henry
Published: February 2007

The majority of, but not all, women with mutations in the BRCAI gene will be affected with breast or ovarian cancer by the age of 70. To establish whether known risk factors modify susceptibility to cancer in these women, we have studied the reproductive histories of 333 North American women who were found by haplotype analysis to carry BRCAI mutations. An increased risk for breast cancer was associated with low parity and with recent birth cohort. The risk of ovarian cancer decreased with increasing age at last childbirth; however, in contrast to the case for sporadic cancer, the risk of ovarian cancer in BRCAI carriers was found to increase significantly with increasing parity. © 1995 Wiley‐Liss, Inc.