bluuebunny/crossref_metadata_2025 huggingface dataset:
Sporadic gastric carcinomas with microsatellite instability display a particular clinicopathologic profile
Authors: Seruca Raquel , Santos Nuno R. , David Leonor , Constáncia Miguel , Barroca Helena , Carneiro Fátima , Seixas Mario , Peltomäki Päivi , Lothe Ragnhild , Sobrinho‐Simöes Manuel
Published: February 2007

Mutations in recently identified genes on chromosomes 2 and 3 seem to be responsible for repair errors (RER+) throughout the genome. This novel genetic mechanism was first reported in hereditary non‐polyposis colorectal cancer syndrome and in cancers that are characteristic of this syndrome, such as carcinomas of the right colon, stomach and endometrium. We investigated the frequency of RER+ phenotype in a series of 34 sporadic gastric carcinomas, in an attempt to see if the RER+ cases displayed any particular morphologic features and/or if they showed distinctive clinicopathologic characteristics. Twelve loci were investigated. We found 23 RER‐ cases (67.6%) and 11 RER+ cases (32.4%). A significant association was found between RER+ carcinomas and localization of the tumors: 9 of the 11 RER+ carcinomas (81.8%) were located at the antrum whereas all the cardiac tumors were RER‐. The RER+ phenotype was also significantly related to the presence of moderate/abundant T‐cell lymphoid infiltration within the tumors. The 3‐year survival rate of patients with RER+ tumors was suggestively longer than that of patients with RER‐ tumors. No significant relationship was found between several clinicopathologic characteristics of the cases, including age, sex, staging, histologic type and ploidy, despite a trend towards an association between RER+ phenotype and advanced age of the patients and poorly differentiated, intestinal type of the carcinomas. The high frequency of microsatellite instability in sporadic gastric carcinomas supports the involvement of this genetic mechanism in gastric carcinogenesis. Gastric carcinomas with the RER+ phenotype tend to occur as poorly differentiated adenocarcinomas in the antrum of elderly patients, display abundant T‐cell infiltration and carry a relatively good prognosis. © 1995 Wiley‐Liss, Inc.
p53 mutations in ovarian tumors, detected by temperature‐gradient gel electrophoresis, direct sequencing and immunohistochemistry
Authors: Kappes Stefanie , Milde‐Langosch Karin , Kressin Philipp , Passlack Babette , Dockhorn‐Dworniczak Barbara , Röhlke Peter , Löning Thomas
Published: February 2007

Samples from 94 ovarian tumors, comprising 24 cystadenomas/adenofibromas, among them 6 benign and 18 borderline tumors, one benign Brenner tumor, 39 carcinomas, 17 sex‐cord stromal tumors, 5 germ‐cell tumors and 8 metastatic or recurrent neoplasms were screened for p53 aberrations by polymerase chain reaction (PCR), temperature‐gradient gel electrophoresis (TGGE), direct sequencing and immunohistochemistry. All sex‐cord stromal and germ‐cell tumors showed wild‐type p53, except for a heterozygous silent germ‐line mutation in one androblastoma. Somatic p53 mutations were detected in only one tumor of the cystadenoma/adenofibroma series (4.2%), in contrast to 38.5% of the carcinomas, among them 57.1% of serous papillary carcinomas, and 12.5 to 22.2% of endometrioid and mucinous carcinomas. By direct sequencing, the mutations of 13 cases were qualified as mis‐sense mutations (n = 10), or I to 2‐bp deletions (n = 3). Only 2 cases were immunohistochemically positive in the absence of detectable p53‐gene abnormalities. The presence of p53 aberrations was significantly correlated with high grade, but not with stage of disease. For 21 bilateral tumors and/or tumors spread to the peritoneum, samples from both ovaries and/or ascites were analyzed. Among these, 16 cases were identical as to the p53 genotype, 5 cases showed discordant p53 states in ovary and/or in ascites DNA. We conclude that somatic p53 mutations are very frequent in serous papillary carcinomas, particularly in tumors of high grade, bilaterality, and peritoneal spread, less frequent in other carcinoma types and extremely rare in borderline and benign tumors of the ovary. © 1995 Wiley‐Liss, Inc.
Tumor uptake of intravenously administered radiolabeled antibodies in ovarian carcinoma patients in relation to antigen expression and other tumor characteristics
Authors: Buist Marrije R. , Kenemans Peter , Molthoff Carla F. M. , Roos Jan C. , Hollander Wim Den , Brinkhuis Mariel , Baak Jan P. A.
Published: February 2007

To study factors that possibly influence the heterogeneous tumor uptake of radiolabeled antibodies, tissues from 34 ovarian‐carcinoma patients were obtained 2 to 8 days after i.v. injection with radiolabeled murine OV‐TL 3 or chimeric MOv 18 (cMOv18). The tumor uptake and the ratio of tumor to normal tissue (T/NT) were studied in relation to the histopathological classification, prior treatment, site of tumor, time interval, antigen expression, volume percentage of (malignant) epithelium in the tumor tissue, and the size of the tumor. The results of immuno‐scintigraphy were also included. In addition, autoradiography using storage phosphor technology was performed on tissue sections from patients injected with iodine‐labeled cMOv18.

Tumor uptake varied largely, not only between patients, but also between tumor deposits within the same patient. Uptake of OV‐TL 3 F(ab')2 was higher than of cMOv18 F(ab')2, but the T/NT ratios were similar. The antibody uptake was positively correlated with the pattern of antigen expression and inversely correlated with the time interval between injection and surgery. No correlation was observed with any of the other factors studied. The visibility with immunoscintigraphy was related to the size of the detected lesion, but not to the other factors studied. Autoradiography showed that antibodies preferentially localized in areas with cancer cells, which were immunohisto‐chemically positive for MOv18. In areas with weak antigen expression, autoradiography revealed less activity. The antigen expression by the tumor is an important factor for estimation of the tumor uptake of radiolabeled antibodies. © 1995 Wiley‐Liss, Inc.
High nm23‐h1/ndpk‐a expression in ewing tumors: Paradoxical immunohistochemical reactivity and lack of prognostic significance
Authors: Aryee Dave N. T. , Ströbel Thomas , Kos Karin , Salzer‐Kuntschik Mechthild , Zoubek Andreas , Veron Michel , Ambros Inge M. , Traincart Francois , Gadner Helmut , Kovar Heinrich
Published: February 2007

Expression of nm23‐HI/NDPK‐A has been reported to correlate inversely with metastasizing potential of rodent experimental cells and some human tumors. In the search for reliable molecular prognostic indicators for Ewing tumors (ET), a group of aggressive presumably neuroectodermal malignancies in children and adolescents, we studied nm23‐HI/NDPK‐A expression. Northern‐blot and RT‐PCR analyses were employed to semi‐quantificatively measure nm23‐HI mRNA levels in ET cell lines and tissue extracts. A panel of monoclonal antibodies (MAbs) were used to evaluate protein abundance by Western blotting and immunohistochemistry. The nm23‐HI/NDPK‐A gene was also investigated on the DNA level to define possible genomic alterations. Our results revealed neither nm23‐HI allelic loss nor gene amplification and failed to show any significant variation in nm23‐HI mRNA or NDPK‐A protein levels of primary or metastatic ET. NDPK‐A protein levels were high and comparable to those of MCF‐7 breast‐cancer cells and to aggressive stage‐IV neuroblastoma cell lines. nm23‐H2/NDPK‐B expression in ET was slightly more variable but generally lower than in MCF‐7 cells. In the immunohistochemical analysis, however, discrepancies in the reactivity patterns with different antibodies were observed. Differential sensitivity to various fixation methods and heat treatment pointed to a structurally polymorphic NDPK‐A protein. nm23‐H1 expression studies using immunohistochemistry for prognostic counselling should thus be interpreted with caution. © 1995 Wiley‐Liss, Inc.
Expression of basic fibroblast growth factor in squamous cell carcinoma of the head and neck is associated with degree of histologic differentiation
Authors: Janot Fraçois , El‐Naggar Adel K. , Morrison Richard S. , Liu Ta‐Jen , Taylor Dorothy L. , Clayman Gary L.
Published: February 2007

Basic fibroblast growth factor (bFGF) is a potent mitogen and angiogenic protein that may function as an autocrine growth regulator in a variety of malignancies. Expression of bFGF in squamous cell carcinoma of the head and neck (SCCHN) was characterized by Western blot and immunohistochemical analyses. We found that the levels of bFGF in tumors were the same or reduced relative to non‐malignant adjacent mucosa. Reverse transcription‐polymerase chain reaction and Southern blot transfer of mRNA derived from 7 SCCHN cell lines showed that the IIIb isoform of FGF‐receptor 2(FGFR2) was expressed at high levels, whereas the IIIC isoform and FGFRI were weakly expressed or not detected. No correlation was observed between levels of bFGF revealed by immunohistochemical staining and vascular counts in frozen sections derived from 11 different SCCHN tumors. Immunohistochemical analysis demonstrated that all differentiated tumors exhibited high levels of bFGF immunoreactivity, while all poorly differentiated tumors exhibited low to nondetectable levels. This expression pattern is consistent with that observed in non‐tumoral mucosa and suggests that other angiogenic factors must play a predominant role in the development of poorly differentiated SCCHN. © 1995 Wiley‐Liss, Inc.
Expression of tumor‐associated 90k‐antigen in human breast cancer: No correlation with prognosis and response to first‐line therapy with tamoxifen
Authors: Foekens John A. , Klun Jan G. M. , Natoli Clara , Van Putten Wim L. J. , Stefano Pia Di , Look Maxime P. , Portengen Henk , Lacobelli Stefano
Published: February 2007

It has been shown that a 90‐kDa protein (90K), with an as yet unknown function, is expressed in the majority of human breast‐cancer tissues. In addition, the serum level of this 90K antigen is elevated in a certain proportion of breast‐cancer patients, and high serum levels are associated with a poor overall survival. It was therefore of interest to determine whether levels of 90K in tumor tissues could be used as a prognostic variable in breast cancer. In the present study, the levels of 90K in primary breast tumor cytosols were studied with respect to the length of relapse‐free or overall survival in 547 patients (median follow‐up, 81.4 months), and the relationship with response to first‐line tamoxifen therapy and the length of progression‐free survival in 184 patients with recurrent disease (median follow‐up, 59.8 months). 90K levels in tumor cytosols were determined with an immunoradiometric assay. The cytosolic contents of 90K were not significantly correlated with age, menopausal status, tumor size, nodal status or differentiation grade. On the other hand, the levels of 90K were positively correlated with those of cytosolic estrogen receptor, progesterone receptor, urokinase‐type plasminogen activator, its inhibitor PAI‐I, cathepsin D and PS2. The cytosolic tumor level of 90K was not associated with the rate of relapse or death in primary breast cancer, nor with response to first‐line therapy with tamoxifen or the length of progression‐free survival in recurrent disease. © 1995 Wiley‐Liss, Inc.
c‐crbB2 gene amplification and protein expression in ovarian epithelial tumors: Evaluation of their respective prognostic significance by multivariate analysis
Authors: Fajac Anne , Benard Jean , Lhomme Catherine , Rey Annie , Duvillard Pierre , Rochards France , Bernaudin Jean‐Francois , Riou Guy
Published: February 2007

c‐erbB2 gene amplification or over‐expression has been reported in ovarian cancer, but their prognostic value remains conflicting. To investigate the respective prognostic significance of c‐erbB2 gene amplification and protein over‐expression, tumor samples were obtained from 65 patients with ovarian adenocarcinoma (9 FIGO stage I, 7 stage ll, 38 stage III and 11 stage IV) followed up for a median period of 71 months. c‐erbB2 gene amplification (>2.5 a.u.) was detected in 9/65 (14%) adenocarcinomas and in none of 5 benign and 8 borderline ovarian epithelial tumors also analyzed. Specimens from 52 of the 65 adenocarcinomas were available for immunohistochemi‐cal analysis. c‐erbB2 protein expression was observed in 23/52 (44%) adenocarcinomas. No correlation was found between c‐erbB2 gene copy number and protein expression. There was no correlation of c‐erbB2 gene copy number or protein expression with any of the clinico‐pathological factors analyzed (i.e., GIGO stage, histological type, histological grade and residual tumor). On univariate analysis, c‐erbB2 gene amplification was associated with poorer survival (p = 0.04). However, in the multivariate analysis of clinico‐pathological factors and c‐erbB2 gene copy number, c‐erbB2 gene amplification did not retain any independent prognostic significance (p = 0.19). No significant survival difference was found between patients with and without c‐erbB2 protein over‐expression in univariate or multivariate analyses. Therefore, neither c‐erbB2 gene amplification nor c‐erbB2 protein over‐expression appears to be a significant prognostic marker in patients with ovarian carcinoma. © 1995 Wiley‐Liss, Inc.
Sequential p53 mutation analysis of pre‐invasive and invasive head and neck squamous carcinoma
Authors: El‐Naggar Adel K. , Lai Syeling , Luna Mario A. , Zhou Xiao‐Dong , Weber Randal S. , Goepfert Helmuth , Batsakis John G.
Published: February 2007

Single‐stranded conformation polymorphism (SSCP) and direct sequencing were performed on uninvolved mucosa, severe dysplasia and invasive carcinoma samples from 20 patients with head and neck squamous carcinoma. Seven (35%) of the non‐invasive lesions and 15 (75%) of the invasive carcinomas manifested p53 mutations. Although the majority of mutations were mis‐sense, resulting in single amino acid substitution, a silent mutation encoding for the same amino acid and 2 non‐sense mutations encoding a stop codon were also observed. Mutations in invasive carcinoma were mostly in exon 8 and involved codons 296, 288 and 298; non‐invasive lesions showed more mutations at exons 5 to 7. Five lesions showed simultaneous mutations in 2 different exons; in 3 both non‐invasive and invasive carcinomas showed primary mutation at exons 5 to 7, and invasive carcinoma showed a secondary mutation at exon 8. Different codon mutations in the same exon between dysplastic and the corresponding carcinoma samples were found in 2 cases. p53 alterations were not observed in any of the normal mucosa samples. No apparent association between p53 mutations and conventional clinicopathologic parameters, including DNA content, was found in this cohort. Our study indicates that (i) p53 alteration is an early event in the genesis of a subset of head and neck squamous carcinomas, (ii) normal mucosa within the resected specimens lacked p53 mutation, (iii) sequential mutations of different exons of the p53 gene suggests accumulation of genetic alterations during the neoplastic transformation of these lesions and (iv) the difference in codon mutation of the same exon between dysplastic and corresponding carcinoma suggests an independent clonal development. © 1995 Wiley‐Liss, Inc.
Detection of TP53 gene mutation in human meningiomas: A study using immunohistochemistry, polymerase chain reaction/single‐strand conformation polymorphism and dna sequencing techniques on paraffin‐embedded samples
Authors: Wang Jia‐Lun , Zhang Zhi‐Jia , Hartman Magdalena , Smits Anja , Westermark Bengt , Muhr Carin , Nistér Monica
Published: February 2007

Mutations in the TP53 tumor suppressor gene have been studied in different types of brain tumors. Little is known about this genetic event in human meningioma, a mostly benign tumor. To investigate the frequency of TP53 gene mutations in human tumors derived from meningeal tissues, paraffin‐embedded tissues from 30 cases (including 2 malignant and 4 atypical meningiomas, as well as 2 hemangioblastomas and 3 hemangiopericytomas) were screened by immunohistochemistry. Polymerase chain reaction/single strand conformational polymorphism (PCR/SSCP) and direct DNA sequencing were thereafter performed in selected cases. Nuclear p53 staining was not seen in any of the 19 benign meningiomas tested, while atypical meningiomas, hemangioblastomas, and hemangiopericytomas displayed nuclear staining in a subpopulation of tumor cells in 4 out of 5, 2 out of 2, and 3 out of 3 cases, respectively. One malignant meningioma showed an intense nuclear staining and a band shift in SSCP. In this case, we identified a mutation in the TP53 gene at codon 161 changing GCC to ACC and resulting in an alteration of alanine to threonine in this position. Our results indicate that TP53 gene mutation may be considered as a marker for malignant transformation in meningioma. p53 immunoreactivity, even in the absence of detectable gene mutation, is also associated with atypia and does not appear in regular benign meningiomas. © 1995 Wiley‐Liss, Inc.
Microsatellite instability in multiple gastric cancers
Authors: Nakashima Hideaki , Honda Masayuki , Inoue Hiroshi , Shibuta Kenji , Arinaga Shinya , Era Shoichi , Ueo Hiroaki , Mori Masaki , Akiyoshi Tsuyoshi
Published: February 2007

To better elucidate the role of genetic instability in the development of gastric cancer, microsatellite alterations were examined in a total of 30 gastric cancers that developed in 14 Japanese patients with multiple gastric cancers, which are considered to have possibly occurred under the same genetic background and in the same microenvironment of the stomach. Microsatellite instability (MSI) in multiple gastric cancers was recognized in 11 out of 14 cases (78.5%) and in 16 out of 30 cancers (53.3%). Eight out of 11 cases showing MSI exhibited a heterogeneity of microsatellite alterations. The incidence of microsatellite instability in the multiple gastric cancers cases was significantly higher than that in the solitary gastric cancer cases reported previously (20.8%: 5 out of 24 cases). These results suggested that (1) genetic instability plays a more important role in the development of multiple gastric cancers than in that of solitary gastric cancer and (2) the heterogeneity of MSI in multiple gastric cancers may not be a rare event, although the significance of the heterogeneity could not be clarified. © 1995 Wiley‐Liss, Inc.
Retinoic acid receptors and retinoid binding proteins in endometrial adenocarcinoma: Differential expression of cellular retinoid binding proteins in endometrioid tumours
Authors: Siddiqui Nadeem A. , Thomas Eric J. , Dunlop William , Redfern Christopher P. F.
Published: February 2007

Retinoic acid is apparently required for the normal differentiation of reproductive epithelium. Cellular abnormalities in retinoid homeostasis could be a factor in the development of endometrial malignancy. We have thus investigated the expression of nuclear retinoic acid receptors (RARs and RXRs) and cellular binding proteins for retinol (CRBP) and retinoic acid (CRABP) in endometrial adenocarcinoma of the endometrioid histological subtype. Ten grade 1, 11 grade 2 and 10 grade 3 tumour samples, as well as 4 samples of severe atypical precan‐cerous endometrial hyperplasia, were studied. No significant difference in expression of RAR‐β was detected in tumour samples compared with normal epithelial cells. RAR‐γ was significantly elevated in grade 1 and 2 carcinomas, but this may be due to greater stromal cell involvement in these lower grade tumours. There was significant elevation of CRBP 1 mRNA in tumour samples. Furthermore, although undetectable in normal endometrial epithelium, CRABP 1 was expressed in 3/11 grade 2 and 9/10 grade 3 carcinomas, with expression being significantly higher where the primary tumour had invaded more than 50% of the total myometrial thickness. Analysis of 2 epithelial‐like endometrial adenocarcinoma cell lines supported the idea that CRABP 1 expression is characteristic of poorly differentiated endometrial adenocarcinoma. Our data suggest that alterations in mechanisms of retinoid homeostasis are a feature of endometrial adenocarcinoma and may contribute to the severity of disease. © 1995 Wiley‐Liss, Inc.
Expression of 2 variant forms of fibroblast growth factor receptor 1 in human breast
Authors: Luqmani Yunus A. , Mortimer Caroline , Yiangou Constantinos , Johnston Caroline L. , Bansal Gurpal S. , Sinnett Dudley , Law Matthew , Charles Coombes R.
Published: February 2007

The expression of variant mRNAs encoding isoforms of fibroblast growth factor receptor (FGFR) 1 with either 2 or 3 Ig‐like loops in the extracellular domain was investigated in human breast tissues and cell lines using a polymerase chain reaction amplification method. Almost all tissues contained both forms of FGFR1, but cancers (n = 137) had a significantly lower proportion of the transcript that encoded the full 3‐loop form compared with non‐malignant biopsies (n = 34). This was confirmed using microdissected populations of normal and cancerous cells from frozen tissue sections. A high ratio of the 2‐to 3‐loop form was found to be predictive of reduced relapsefree survival. In both groups, however, the predominant form of FGFR1 was that encoding the 2‐loop receptor. Cell lines derived from a variety of tissues, including breast, also co‐expressed both variants of FGFR1, suggesting their presence within the same cell type. Again, there was a similar preponderance of the shorter isoform. Our results were confirmed at the protein level, where out of 5 cancers analysed 4 expressed more of the 2‐loop form than the 3‐loop form. Our findings suggest that cells may normally simultaneously express several splice variants of FGFR1, and aberrant expression or a change in their relative amounts (i.e., in malignancy) could contribute to modified responses to either autocrine or paracrine factors. © 1995 Wiley‐Liss, Inc.
ERBB2 gene amplification detected by fluorescent differential polymerase chain reaction in paraffin‐embedded breast carcinoma tissues
Authors: An Han‐Xiang , Niederacher Dieter , Beckmann Matthias W. , Göhring Uwe‐Jochen , Scharl Anton , Picard Frauke , van Roeyen Claudiavan , Schnürch Hans‐Georg , Bender Hans G.
Published: February 2007

For quantificative determination of ERBB2 gene amplification in archival human carcinoma specimens we have developed a rapid, non‐radioactive approach, which is based on the differential polymerase chain reaction (PCR) and fluorescent DNA technique. Sequences from the ERBB2 gene and from a singlecopy reference gene were amplified simultaneously by PCR, in which one of each primer pair was fluorescently labelled. PCR products were separated by polyacrylamide gel electrophoresis in an automated DNA sequencer and directly quantified after laser activation and emission scanning using appropriate software. This fluorescent differential polymerase chain reaction (fd‐PCR) method was used for quantificative determination of ERBB2 gene amplification in 195 formalin‐fixed, paraffin‐embedded breast carcinoma tissues. ERBB2 gene amplification was found in 52 (26%) of these tumors and correlated significantly with tumor size, absence of estrogen receptor (ER) and pS2 expression, but not with absence of progesterone receptor (PR) or presence of epidermal growth factor receptor (EGF‐R) expression, lymph‐node metastases or grading. In univariate analysis, ERBB2 gene amplification showed no significant correlation with clinical outcome, either in the whole population or in the subgroup defined by positive axillary lymph‐node metastases. However, within the node‐negative subgroup, patients with ERBB2 gene amplification had significantly decreased relapse‐free survival and overall survival (p < 0.05). The fd‐PCR assay is a valuable tool for determination of amplification of ERBB2 gene as well as further oncogenes. In this way, more detailed information about individual tumor biology may be acquired by a routine assay. © 1995 Wiley‐Liss, Inc.
Expression of the MAGE gene family in human head‐and‐neck squamous‐cell carcinomas
Authors: Eura Masao , Ogi Kojiro , Chikamatsu Kazuaki , Lee Kang Dac , Nakano Koji , Masuyama Keisuke , Itoh Kyogo , Ishikawa Takeru
Published: February 2007

The MAGE genes encode certain tumor‐associated antigens recognized by cytotoxic T lymphocytes. We investigated the expression of the MAGE‐1, ‐2, ‐3, ‐4, ‐41 and ‐6 genes in 88 head‐and‐neck squamous‐cell carcinomas (83 fresh tumor samples and 5 cell lines), using a reverse‐transcription‐polymerase‐chain‐reaction assay, followed by dot‐blot hybridization with sequence‐specific oligonucleotides and/or restriction‐enzyme‐pattern analysis. The MAGE‐1, ‐2, ‐3, ‐4, ‐41 and ‐6 genes were expressed at the mRNA level in 27, 34, 36, 22, 16 and 35, respectively, of 83 fresh tumor samples. At least one of these genes was expressed in 59 of the 83 samples. Neither non‐tumor inflammatory cells nor normal tissues were positive for these genes. The MAGE‐1 gene was expressed relatively frequently in SCC of the oropharynx, hypopharynx and maxillary sinus, but at lower rates in SCC of the larynx and of the tongue and oral cavity. MAGE‐1 was frequently expressed in poorly differentiated SCC, somewhat less frequently in moderately differentiated SCC, and only infrequently in well‐differentiated SCC. The expression levels of the other MAGE genes also varied with the anatomic site as well as the degree of differentiation. Our results suggest that specific immunotherapy against MAGE gene products may be useful for patients with head‐and‐neck carcinomas. © 1995 Wiley‐Liss, Inc.
Expression of autocrine motility factor receptor in human esophageal squamous cell carcinoma
Authors: Maruyama Kentaro , Watanabe Hideomi , Shiozaki Hitoshi , Takayama Takuya , Gofuku Junji , Yano Hiroshi , Inoue Masatoshi , Tamura Shigeyuki , Raz Avraham , Monden Morito
Published: February 2007

Autocrine motility factor and its receptor (gp78) have been shown to play an important role in tumor cell migration, invasion and metastasis. We have detected gp78 expression in buffered‐formalin‐fixed, paraffin‐embedded sections of esophageal squamous cell carcinomas using an anti‐gp78 monoclonal antibody (MAb), 3F3A, and examined the relationship between gp78 expression and clinicopathological and prognostic factors. In 55 of 101 (54%) patients, gp78 was detected in the tumor cells. The frequency of gp78‐positive expression was significantly associated with tumor size, infiltrative growth, depth of invasion and lymph node metastasis. The cumulative survival rate of patients with gp78 was significantly lower than that of patients without gp78. Our results suggest that autocrine motility factor receptor (gp78) expression could be a useful biomarker for malignancy grading and prognosis in patients with esophageal squamous cell carcinoma. © 1995 Wiley‐Liss, Inc.
Microsatellite instability in oral cancer
Authors: Ishwad Chandramohan S. , Ferrell Robert E. , Rossie Karch M. , Appel Billy N. , Johnson Jonas T. , Myers Eugene N. , Law John C. , Srivastava S. , Gollin Susanne M.
Published: February 2007

Generalized genomic instability, detected as somatic changes in allele sizes at microsatellite loci in tumors compared to peripheral lymphocyte DNA, is a recently recognized mechanism of mutation in cancer. Such instability results from the Somatic loss of DNA mismatch repair capability. Germ‐line mutations at DNA mismatch repair loci confer susceptibility to colon cancer in hereditary non‐polyposis colorectal cancer. Somatic loss of DNA mismatch repair has been reported in a large variety of other tumor types. Our goal was to determine the frequency of microsatellite instability in a large series of oral tumors. Out of 91 tumors analyzed for microsatellite instability, 6 (7%) showed microsatellite instability. Instability was observed at multiple loci with a range of 50‐74% of loci affected. Alterations include both increase (74%) and decrease (26%) in allele sizes. The proportion of alleles affected ranged from 30‐58% of all alleles. Our data suggest that somatic genomic instability plays a role in the pathogenesis of a small subset of oral tumors. © 1995 Wiley‐Liss, Inc.
Activation of the precursor of gelatinase A/72 kda type IV collagenase/MMP‐2 in lung carcinomas correlates with the expression of membrane‐type matrix metalloproteinase (MT‐MMP) and with lymph node metastasis
Authors: Tokuraku Masato , Sato Hiroshi , Murakami Sinya , Okada Yasunori , Watanabe Yoh , Seiki Motoharu
Published: February 2007

We have identified a novel membrane‐type matrix metalloproteinase (MT‐MMP) expressed on the cell surface and inducing activation of pro‐gelatinase A in vitro. In this study, we further examined the possibility that MT‐MMP is the activator of pro‐gelatinase A in tumors as well as in vitro. Expression of MT‐MMP mRNA was analyzed by Northern blotting in 58 cases of human lung carcinomas. MT‐MMP mRNA expression was increased in tumor tissues compared with adjacent normal tissues. The ratio of MT‐MMP mRNA levels in tumor/normal tissues (T/N ratio) was 3.19 ± 1.62 in 29 cases of adenocarcinoma, 3.09 ± 1.44 in 24 cases of squamous cell carcinoma, 4.40 ± 0.47 in 3 cases of large cell carcinoma and 3.63 ± 2.11 in 2 cases of small cell carcinoma, respectively. Activated gelatinase A, as detected by gelatin zymography, was also predominant in tumors compared with normal tissue counterparts, though the difference in mRNA levels was not significant. The activation ratio of gelatinase A in tumor vs. normal tissues correlated well with that of MT‐MMP mRNA expression and with lymph node metastases. Our findings suggest that MT‐MMP is indeed the tumor‐specific activator of pro‐gelatinase A in lung carcinomas and is important to initiate invasion of basement membranes. © 1995 Wiley‐Liss, Inc.
Microsatellite instability and alterations in the HMSH2 gene in human ovarian cancer
Authors: Fujita M. , Enomoto T. , Yoshino K. , Nomura T. , Buzard G. S. , Inoue M. , Okudaira Y.
Published: February 2007

The role of the replication error‐positive (RER+) phenotype in the development of specific subtypes of sporadic ovarian carcinomas was examined by screening for the presence of microsatellite instability (MI) in 47 tumors. The overall frequency of ovarian MI was 17% only. However, MI occurred in 50% of the ovarian endometrioid‐type tumors, which was significantly more often than in all the other histological sub‐types combined (8%). Five of the 8 RER+ tumors exhibited most marked type I instability, possibly representing a different mechanism than for the remaining type 2 tumors. The cDNA of the mutation suppression gene hMSH2, the gene most often associated with MI, was screened for alterations in 8 MI‐positive and 5 MI‐negative ovarian tumors. Only 3 changes were found. Complete loss of hMSH2 mRNA expression was detected in 1 tumor, while another expressed only an abnormal transcript containing a deletion of exon 3. One additional RER+ serous adenocarcinoma contained a rare polymophism with a non‐conservative amino acid change. One of 8 RER+ tumors showed loss of heterozygosity at the hMSH2 loci. Genetic instability, caused in part by alterations in the hMSH2 gene, may play an important role in the sporadic endometrioid subtype of ovarian tumors. Other mutator‐phenotype genes may be responsible for the remaining cases of RER+ ovarian tumors. © 1995 Wiley‐Liss, Inc.
Detection of p53 gene alteration in renal‐cell cancer by micropreparation techniques of tumor specimens
Authors: Kuczyk Markus A. , Serth Jürgen , Bokemeyer Carsten , Jonassen Jens , Arndt Heike , Paeslack Ute , Werner Martin , Tan Hong‐Ki , Jonas Udo
Published: February 2007

Alterations in the p53 tumor‐suppressor gene have been identified in a variety of human malignancies, including renalcell cancer. A technique for the isolation of tumor areas from tissue specimens to analyze formalin‐fixed and paraffinembedded tumors and try to avoid a disturbance of the results due to genetic background signal by the presence of tumorinfiltrating lymphocytes, was established. The presence of lymphocytes within the tumor areas investigated was determined by immunohistochemical staining for CD3, a lymphatic surface antigen. Following the isolation of about 100‐200 tumor cells, PCR‐directed molecular genetic analysis was performed. A highly informative allelotyping approach for the detection of loss of heterozygosity (LOH), determining BstUI‐ and VNTR‐polymorphisms, a 100‐bp marker directly localized in intron I of the p53 gene, as well as screening for mutations by single‐strand conformation polymorphism analysis (SSCP) in exons 5‐8, were used. Out of 44 renal‐cancer specimens, 33 (75%) were informative for PCR‐directed RFLP‐analysis. Allelic loss at the p53 gene locus was observed in 10 of 33 cases (33%). No correlation between p53 gene alteration and T‐stage, histological grade or histological differentiation could be observed. Alterations in the p53 gene, as detected by a molecular genetic as well as an immunohistochemical approach, were correlated to overall survival. During univariate analysis histological grade, lymphnode status and the presence of distant metastases could be identified as prognostic parameters for overall survival. During multivariate analysis none of the factors investigated remained an independent prognosticator for survival. Summarizing these results, it seems unlikely that p53 gene alterations will serve as an important new factor for the clinical prognosis of patients with renal‐cell cancer. © 1995 Wiley‐Liss, Inc.
Expression of E‐ and N‐cadherin in renal cell carcinomas, in renal cell carcinoma cell lines in vitro and in their xenografts
Authors: Tani Taneli , Laitinen Liisa , Kangas Lauri , Lehto Veli‐Pekka , Virtanen Ismo
Published: February 2007

E‐ and N‐cadherins are proteins involved in intercellular adhesion and are localized, e.g., in the adherens junctions of epithelial cells. Kidney tubules express these molecules in a distinctive pattern, the expression of N‐cadherin being restricted to proximal tubules and that of E‐cadherin to distal tubules and collecting ducts. Renal cell carcinomas (RCCs) and oncocytomas are considered to originate from these tubular epithelia. To find out whether cadherins could serve as markers for a cellular origin of these tumors, we studied the expression of E‐ and N‐cadherins in RCCs and oncocytomas, in cell lines derived from RCCs as well as in tumors grown in nude mice. Most RCCs co‐expressed E‐ and N‐cadherins, as did 2 of the 4 cell lines studied. The expression pattern did not correlate with the histological grade of the tumors, and even the least differentiated tumors, as well as metastases, showed expression of cadherins. Renal oncocytomas expressed E‐cadherin but not N‐cadherin, which is in line with previous studies that have proposed a collecting duct origin for these tumors. Papillary renal neoplasms, a separate entity usually not classified as RCC, expressed neither of the cadherins studied despite the abundant expression of β‐catenin. Our results suggest that most RCCs co‐express the characteristic adhesion molecules of both proximal and distal tubules, which makes it questionable whether the origin of these tumors can be reliably located to any distinct part of the renal tubule. Our results also suggest that in RCCs the increased histological grade is not directly associated with changes in the expression of either of the cadherins, indicating other mechanisms underlying the deficient capacity to form polarized tubular structures. © 1995 Wiley‐Liss, Inc.
Application of a new bioresorbable film to guided bone regeneration in tibia defect model of the rabbits
Authors: He Hong , Huang Jianqi , Chen Guanfu , Dong Yan
Published: February 2007

Aim is to study the effect of calcium alginate film (CAF) on guided bone regeneration (GBR). Circular bone defects with 5 mm diameter were created in both tibias in 60 rabbits. The defects covered with CAF served as the experimental site, and with collagen membrane (CM) or with no membrane both served as the control site. Healing was analyzed by gross, X‐ray, electromicroscopy, histology, immuno‐histochemical studies, and an image pattern analysis system after 1, 2, 4, 6, and 8 weeks. The CM control sites showed more macrophages, and CM were absorbed more slowly while collecting fewer osteoinductive factors (p < 0.01) in the early weeks. CAF induced dense bone formation, whereas CM induced less new bone, and the blank control sites effected the worst. In conclusion, the effect of CAF group gave better results than blank control group and CM group on GBR in this animal model. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007
Interleukin‐6 (IL‐6) and low O2 concentration (1%) synergize to improve the maintenance of hematopoietic stem cells (pre‐CFC)
Authors: Kovačević‐Filipović Milica , Petakov Marijana , Hermitte Francis , Debeissat Christelle , Krstić Aleksandra , Jovčić Gordana , Bugarski Dijana , Lafarge Xavier , Milenković Pavle , Praloran Vincent , Ivanović Zoran
Published: February 2007

Low O2 concentration (1%) favors the self‐renewal of hematopoietic stem cells and inhibits committed progenitors (CFC). Since IL‐6 influences both stem cells and committed progenitors at 20% O2, we studied its effects in cultures at 1% O2. The pre‐CFC activity in Lin− population of mouse bone marrow was analyzed following 10 days of serum‐free culture in medium (LC1) supplemented with IL‐3 with and without IL‐6, at 20 and 1% O2 and phenotypic differentiation and proliferative history monitored. The IL‐6 receptor expression and initiation of VEGF‐A synthesis were also investigated. At 20% O2, the effects of IL‐6 on pre‐CFC were negligible but effects on CFC were apparent; conversely, at 1% O2, the IL‐6 enhances activity of pre‐CFC but not of CFC. Unlike at 20% O2, at 1% O2 a subpopulation of cells remained Lin− in spite of extensive proliferation. However, the absolute number of Lin− cells, did not correlate with pre‐CFC activity. A relative increase in VEGF transcripts at 1% O2 in presence of IL‐3 alone was enhanced by the addition of IL‐6. IL‐6 enhanced pre‐CFC activity at 1% O2 and this was correlated to the induction of VEGF. These data reinforce the concept that physiologically low oxygenation of bone marrow is a regulator of stem cell maintenance. Since the 20% O2 does not exist in tissues in vivo, further studies in vitro at lower O2 concentrations should revise our knowledge relating to cytokine effects on stem and progenitor cells. J. Cell. Physiol. 212: 68–75, 2007. © 2007 Wiley‐Liss, Inc.
Endothelin induces functional hypertrophy of peritubular smooth muscle cells
Authors: Romano Francesca , Gambara Guido , De Cesaris Paola , Ziparo Elio , Palombi Fioretta , Filippini Antonio
Published: February 2007

When chronically stimulated with agonists of contraction, smooth muscle cells (SMCs) undergo cell hypertrophy, a process defined as increase in size and potentiation of the contractile phenotype in the absence of proliferation. Hypertrophic response has long been associated to a number of pathologies of the cardiovascular and respiratory systems. We have investigated the phenotypic and functional response of SMCs to long‐term treatment with endothelin. Our model was primary cultures of peritubular smooth muscle cells (PSMC) a testicular cell type target of locally produced endothelin and characterized by an unusual phenotypic stability when cultured in simple medium in complete absence of serum. We report the following responses of PSMC to 4‐day exposure to ET‐1: (i) increased protein synthesis without induction of cell proliferation; (ii) increase in cell size (evaluated by means of flow cytometry) and increased expression of SM‐α‐actin, desmin, caldesmon and calponin, markers of the contractile phenotype. In experiments of selective stimulation of either ETA or ETB receptor subtypes, both proved to be involved in inducing the observed hypertrophic responses. The hypertrophic cells exhibit the ultrastructural features of differentiated SMCs and are capable of calcium mediated contractile response when acutely stimulated with ET‐1 specifically through ETA and/or ETB receptors, as evaluated by calcium imaging and scanning electron microscopy. These observations demonstrate that engagement of ET receptors is capable of inducing potentiation of the contractile phenotype and functional hypertrophy of PSMC. J. Cell. Physiol. 212: 264–273, 2007. © 2007 Wiley‐Liss, Inc.
MnSOD mimic compounds can counteract mechanical stress and islet β cell apoptosis, although at appropriate concentration ranges
Authors: Pedullà Marcella , d'Aquino Riccardo , Desiderio Vincenzo , de Francesco Francesco , Puca Andrew , Papaccio Gianpaolo
Published: February 2007

Pancreatic islets are commonly isolated for research and transplantation without taking into consideration that they undergo mechanical or chemical stress during this process. In order to counteract both types of injuries, the compound AEOL10150, a novel MnSOD mimic, was added during isolation of islet at concentrations ranging from 18 to 100 µM. Mechanical or chemical stress‐related pro‐apoptotic signals were then studied. We demonstrate that this MnSOD mimic diminishes the negative effects of mechanical stress by blocking insulin impairment, production of non‐specific islet β‐cell proteins, transcription of iNOS and FAS, activation of caspase‐3 and ‐9 and, ultimately, apoptosis. Moreover, the effects of the MnSOD mimic on isolated islets were greatly influenced by dosage: the best dose able to fully counteract mechanical stress was found to be 100 µM; doses ≥150 µM were themselves highly toxic for islet cells. On the other hand, rIL‐1β‐induced chemical stress is rather complex, and there was no protection in this scenario. Therefore, contrarily to what has been previously reported, MnSOD mimic administration is only capable of counteracting mechanical stress, and not cytokine‐induced cytotoxicity, and that this drug acts within a limited concentration range. J. Cell. Physiol. 212: 432–438, 2007. © 2007 Wiley‐Liss, Inc.
Infection and direct injury in human hepatocyte explants and a hepatoblastoma cell line due to hepatiticomimetic (non‐hepatitis) viruses
Authors: Phromjai Jurairatana , Aiba Nobuyasu , Suzuki Mikiko , Sato Hitoshi , Takahara Terumi , Kondo Sachiko , Shiraki Kimiyasu
Published: February 2007

Hepatitis is caused by hepatitis viruses, but hepatitis or hepatocellular enzyme abnormalities is sometimes associated with infection by the hepatiticomimetic viruses. The direct and indirect effects of infection with hepatiticomimetic viruses were examined in two human hepatocyte systems. Poliovirus, adenovirus, and herpes simplex virus (HSV) induced cytopathology in Hep G2 cells. Measles virus caused no change in hepatocytes. Poliovirus infection did not affect cellular protein synthesis, and the peak of hepatocellular enzyme release coincided with the peak of virus release. The increase in adenovirus protein synthesis correlated with the decrease of transferrin synthesis, and enzyme release was not prominent. HSV induced viral protein synthesis with enhanced processing and inhibition of synthesis of α1‐antitrypsin. The peak of enzyme release was later than the peak of virus release. In primary hepatocytes, poliovirus, adenovirus, and induced extensive cytopathology and enzyme release, and VZV caused cytopathology and significant but minute enzyme release. The ratio of lactate dehydrogenase to aspartate aminotransferase release was larger in poliovirus infection in both hepatocytes than in HSV or VZV infection. Although poliovirus and adenovirus are released by cytolysis and HSV and VZV are secreted by exocytosis of cytoplasmic vacuoles, enzyme release was independent of the type of virus release. Adenovirus showed strong cytotoxicity but did not modify the membrane nor cause enzyme release. Enzyme release was associated with modification of the surface membrane due to apoptosis with poliovirus and necrosis with HSV. Consequently hepatocellular injury by viral infection did not reflect the amount or pattern of hepatocellular enzyme release. J. Med. Virol. 79:413–425, 2007. © 2007 Wiley‐Liss, Inc.
HPV16 E6 variants and HLA class II polymorphism among Chinese women with cervical cancer
Authors: Wu Yuping , Liu Benrong , Lin Wenzhi , Xu Yunping , Li Longyu , Zhang Yanling , Chen Shangwu , Xu Anlong
Published: February 2007

Infection with human papillomavirus type 16 (HPV16) confers a high risk for the development of cervical cancer. Variants of this virus may interact differentially with host genetic factors, possibly affecting the disease pathogenesis. This study was designed to investigate the association between HPV16 E6 variants and human leukocyte antigen (HLA) polymorphism within a Chinese population. Peripheral blood from HPV16 positive Chinese women with cervical carcinoma, who had previously been tested for HPV16 E6 variants, was used for HLA class II typing. It was found that there was a significant positive association between DQB1*060101 allele and HPV16 As variant‐positive cervical cancers (OR, 4.47; Pc = 0.0018). A negative relationship was found between DRB1*150101‐DQB1*0602 haplotype and decreased risk for HPV16 As variant‐positive cervical cancers (OR = 0.31; P = 0.037). Similar tendency was observed for the haplotype DRB1*070101‐DQB1*0201 with HPV16 As variant‐positive cervical cancers (OR = 0.16, P = 0.024). Additionally, as for the HPV16 E6 prototype‐positive cervical cancers, a significant positive association was found in DQB1*060101 allele (OR = 5.95; P = 0.002; Pc = 0.036), and similar trends were observed for DQB1*030201 (OR = 10.87, P < 0.0001; Pc = 0.0018), and DPB1*1301(OR = 7.40, P = 0.002; Pc = 0.04). It was found that there was no significant association between DRB1‐DQB1 haplotype and HPV16 prototype‐positive cervical cancers. These data indicate that host genetic factors, such as HLA polymorphism, may determine the potential oncogenicity of the HPV16 E6 variant. The results suggest that a specific match between E6 variant proteins and HLA class II alleles may contribute to HPV16‐related cervical carcinogenesis in a certain Chinese population. J. Med. Virol. 79:439–446, 2007. © 2007 Wiley‐Liss, Inc.
Comparison of hepatitis B virus subgenotypes in patients with acute and chronic hepatitis B and absence of lamivudine‐resistant strains in acute hepatitis B in Japan
Authors: Hayashi Kazuhiko , Katano Yoshiaki , Takeda Yasushi , Honda Takashi , Ishigami Masatoshi , Itoh Akihiro , Hirooka Yoshiki , Nakano Isao , Yano Motoyoshi , Goto Hidemi , Yoshioka Kentaro , Toyoda Hidenori , Kumada Takashi
Published: February 2007

Hepatitis B virus (HBV) has been classified into eight genotypes and can be further divided into several subgenotypes that have different geographic distributions. Because of increased human migration, the prevalence of rare subgenotypes is increasing in Japanese patients with acute hepatitis B. Lamivudine‐resistant strains of HBV have begun to emerge in association with chronic hepatitis B. The aim of this study was to investigate the distribution of HBV subgenotypes and lamivudine‐resistant strains in patients in Japan with acute hepatitis B. One hundred twenty‐three patients with acute hepatitis B and 123 with chronic hepatitis B were studied. HBV subgenotypes and lamivudine‐resistance mutations were determined by direct sequencing of the preS and polymerase region, respectively. HBV subgenotypes Aa (n = 3), Ae (n = 23), Ba (n = 7), Bj (n = 3), Cs (n = 7), Ce (n = 76), D (n = 2), and H (n = 2) were detected in patients with acute hepatitis. In patients with chronic hepatitis, HBV subgenotypes Ae (n = 4), Ba (n = 1), Bj (n = 18), and Ce (n = 100) were found. Non‐common Japanese subgenotypes, that is, non‐Bj and non‐Ce, were detected more frequently in patients with acute hepatitis (35.8%) than in patients with chronic hepatitis (4.1%) (Odds ratio, 0.076; 95%CI, 0.029–0.200; P < 0.0001). Lamivudine‐resistance mutations were detected in chronic hepatitis patients with breakthrough hepatitis but not in other patients. In conclusion, the prevalence of uncommon Japanese HBV subgenotypes is expected to increase, although lamivudine‐resistant strains have not yet been found in patients with acute hepatitis B. J. Med. Virol. 79:366–373, 2007. © 2007 Wiley‐Liss, Inc.