bluuebunny/crossref_metadata_2025 huggingface dataset:
The effect of E1A transfection on MMP‐9 expression and metastatic potential
Authors: Bernhard Eric J. , Hagner Beverly , Wong Christopher , Lubenski Irena , Muschel Ruth J.
Published: February 2007

The expression of MMP‐9 in rat embryo fibroblasts (REF) transformed with Ha‐ras or with Ha‐ras and v‐myc is associated with metastatic behavior. In contrast, REF transformed with Ha‐ras and the adenovirus EIA genes (EIA) are tumorigenic, do not release MMP‐9 and are rarely metastatic. In this report, we establish that EIA expression results in decreased levels of MMP‐9 mRNA in an Ha‐ras and v‐myc transformed cell line and examine which of the functional domains of EIA participate in the inhibition of MMP‐9 expression and which contribute to the suppression of metastasis. The metastatic 2.10 REF line, derived by co‐transfection with v‐myc and Ha‐ras, constitutively expresses high levels of MMP‐9 (92 kDa gelatinase). Transfection of E1A wild‐type plasmids into this cell line eliminates detectable MMP‐9 mRNA expression and greatly reduces MMP‐9 activity. Transfection of 2.10 with E1A plasmids encoding mutations in conserved region 2 (CR2) retained inhibition of MMP‐9 similar to the inhibition seen with wild‐type E1A. Transfection with E1A containing mutations in CRI or the amino terminal region diminished, but did not fully inhibit, MMP‐9 expression. In contrast, inhibition of MMP‐9 was lost in with E1A mutations in CR3. Cells transfected with E1A mutants in CRI, the amino terminal region or CR3 retained metastatic behavior. Our Findings delineate the regions of E1A responsible for MMP‐9 inhibition and further define the domains of E1A responsible for inhibition of metastasis.
Kinetics of cisplatin‐induced apoptosis in murine mammary and ovarian adenocarcinomas
Authors: Meyn Raymond E. , Stephens L. Clifton , Hunter Nancy R. , Milas Luka
Published: February 2007

There is mounting evidence to indicate that the mode of cell death known as apoptosis plays an important role in cancer therapy. Most supporting observations have come from experiments conducted in vitro, and it is important to extend such studies to in vivo systems. We have therefore evaluated the magnitude and kinetics of apoptosis induction in tumors from mice treated with cisplatin (CP). Two transplantable murine tumors were studied: a mammary adenocarcinoma, MCa‐4, and an ovarian adenocarcinoma, OCa‐l. Tumor‐bearing mice were injected with various doses of CP, and specimens were obtained over several days. Apoptosis was scored by morphometric analysis of histological sections of the tumors using the features characteristic of cells undergoing this mode of cell death. The results showed a significant apoptotic response in both tumors within a few hours after injection of the drug. The kinetics were very broad, with apoptotic cells present over essentially the entire time course studied. Dose‐response relationships for CP‐induced apoptosis were compared to the tumor response measured in terms of tumor growth delay.
Attributable risks for stomach cancer in Northern Italy
Authors: Vecchia Carlo La , D'Avanzo Barbara , Negri Eva , Decarli Adriano , Benichou Jacques
Published: February 2007

The proportions of gastric cancer cases attributable (or attributable risks, AR) to consumption of traditional foods (i.e., pasta, rice and maize), low intake of betacarotene and vitamin C, short duration of use of an electric refrigerator, low educational level, and family history of gastric cancer were computed using data from a casecontrol study conducted in Northern Italy. Between 1985 and June 1993 a total of 746 incident, histologically confirmed gastric cancer cases and 2,053 controls admitted to the same network of hospitals for acute, nonneoplastic, nondigestivetract diseases, unrelated to longterm modifications of diet, were interviewed. The ARs were 48% for low intake of betacarotene, 40% for high consumption of traditional foods, and 16% for low intake of vitamin C. Overall, these 3 dietary factors explained 73% of the gastric cancer cases in the population. Five percent of all cases were attributable to less than 30 years' use of an electric refrigerator, 15% to low educational level, and 5% to family history of gastric cancer. In individuals over age 60, a greater proportion of cases was attributable to traditional foods, low education and late adoption of electric refrigeration (58% vs. 32% aged under 60), suggesting that correlates of lower social class, influenced lifestyle and dietary habits more markedly in earlier than in more recent generations. According to our estimates, over 3 quarters of the gastric cancer cases in this area are explainable in terms of the risk factors considered. Increased consumption of vitamin C and betacarotene, and reduced consumption of traditional foods, would help to avoid over 10,000 out of 14,000 stomachcancer deaths in Italy every year. Consequently, stomach cancer, which is still the third leading cause of cancer death in Italy, would represent only about 2% of all cancer deaths. © 1995 Wiley‐Liss, Inc.
Receptor tyrosine kinases expressed in metastatic colon cancer
Authors: Craven Rolf J. , Xu Lihui , Weiner Timothy M. , Fridell Yih‐Woei , Dent Georgette A. , Srivastava Sudhir , Varnum Brian , Liu Edison T. , Cance William G.
Published: February 2007

Using a PCR‐based cloning technique, we have isolated a series of DNA fragments coding for tyrosine kinases that are expressed in a metastatic human colon tumor, and have subsequently analyzed their expression pattern at the protein level in human tumors. We identified both the α and the β forms of the platelet‐derived growth factor receptor (PDGFR), axl and 8 other genes, including 3 cytoplasmic tyrosine kinases. To study their expression in human colon cancer, we performed Western blots of matched sets of normal tissues and of carcinomas from the same patient. These revealed that the α‐PDGFR migrates predominantly as a 200‐kDa band in 8/8 normal tissues, and as a 170‐kDa band in 17/17 malignant tissues, as well as in colonic polyps, suggesting that expression of an isoform of this receptor may be a marker for the progression of colon cancer. Additional studies showed that the Axl receptor tyrosine kinase was expressed at 10‐fold higher levels in a peritoneal metastatic nodule than in other normal and malignant tissues. Immunohistochemistry revealed Axl over‐expression specifically in the malignant cells of the tumor. This indicates that over‐expression and possibly a differential processing event of tyrosine kinase receptors may be involved in colon cancer, and that they are potential markers for the progression of this disease. © 1995 Wiley‐Liss. Inc.
NM23 gene expression correlates with cell growth rate and S‐phase
Authors: Caligo Maria A. , Cipollini Giovanna , Fiore Lisa , Calvo Simonetta , Basolo Fulvio , Collecchi Paola , Ciardiello Fortunato , Pepe Stefano , Petrini Mario , Bevilacqua Generoso
Published: February 2007

Two human NM23 genes have been identified: NM23.HI and NM23.H2 coding for the A and B subunit of a nucleoside diphosphate kinase (NDPK), respectively. NM23.HI gene has been proposed as a suppressor of metastatic ability in tumor cells, NM23.H2 is identical to the c‐myc transcription factor, PuF. The NM23 coding sequence is strongly preserved through different species. Indirect evidence of various types has been accumulated and seems to support an implication of NM23 in cell proliferation. This report shows that the NM23 gene expression is strictly related to the growth state of the cells. Two different in vitro systems (human peripheral blood lymphocytes and human breast epithelial cell line MCF‐ 10A) and one in vivo (human primary infiltrating ductal breast carcinomas) system have been investigated. The mRNA is present in PHAstimulated peripheral blood lymphocytes, whereas it is nearly undetectable in their resting counterparts. The level of the NM23 gene expression parallels the fraction of cells incorporating thymidine (S‐phase) in neoplastic mammary tissues. In synchronously cycling MCF‐10A cells NM23.HI mRNA reaches a maximum abundance in the S‐phase and is absent or only present at very low levels during G0/G1 phase, whereas NM23.H2 is present in growth‐ arrested cells but is upregulated following serum growth stimulation.
Alternative splicing of the neurofibromatosis 1 gene correlates with growth patterns and neuroendocrine properties of human small‐cell lung‐carcinoma cells
Authors: Koh Takashi , Yokota Jun , Ookawa Keizo , Kina Tatsuo , Koshimura Kunio , Miwa Soichi , Ariyasu Tetsuya , Yamada Hitoshi , Osaka Mitsuhiko , Haga Hironori , Hitomi Shigeki , Sugiyama Taketoshi , Takahashi Rei
Published: February 2007

Two distinct transcripts, type I and type II, of the neurofibromatosis I (NF) gene are generated by alternative splicing in the region corresponding to the gene's GTPase‐activating protein‐related domain (GRD). Relative expression levels of these 2 transcripts were previously correlated to neural differentiation. Since small‐cell lung carcinoma (SCLC) often exhibits neuroendocrine properties, we analyzed the type‐l to type‐ll mRNA ratio in 15 SCLC cell lines, using reverse transcriptase and polymerase chain reaction methods. The type‐l mRNA was predominant in 10 cell lines; 8 of them grew as floating aggregates in culture and had high L‐dopa decarboxylase (DDC) activity. The other 5 lines predominantly expressed type‐ll mRNA, adhered to the culture substrate, and expressed low or undetectable levels of neural cell‐adhesion molecule (NCAM) antigen and DDC activity. N2 + one of the subclones of NCI‐N417 cells, exhibited a higher type‐l to type‐ll ratio after the cells had adhered to a laminin‐coated plate and had emitted neurite‐like processes. These findings provide evidence that alternative splicing patterns of NFI mRNA correlate with the mechanisms that regulate the growth patterns and neuroendocrine properties of SCLC cells in vitro.
Reduced tumorigenicity of threshold syngeneic tumor inocula in XID‐bearing mice treated with natural antibodies
Authors: Chow Donna A.
Published: February 2007

Several correlative experimental approaches have provided evidence that polyclonal serum natural antibodies (NAb) participate in the defense against small syngeneic tumor foci in vivo. The threshold subcutaneous tumor inoculum model of incipient neoplasia has consistently revealed an inverse relationship between tumorigenicity in vivo. and the NAb binding capacity of the tumor injected or the anti‐tumor NAb levels of the recipient animals, including B cell—deficient mice bearing the xid mutation of the CBA/N mouse strain. Now passive i.v. administration of whole normal syngeneic serum NAb in bolus injections, given I each day beginning on day ‐2, ‐ I or 0 prior to the threshold s.c. challenge of xid‐bearing CBA/N or male (CBA/N × CBA/ J)F1 mice with syngeneic RI‐28 lymphoma cells, consistently and significantly reduced their tumorigenicity assayed as tumor appearance and latency. Three similar injections of an ammonium sulfate—precipitated fraction of whole serum NAb also reduced tumor frequency and latency, while a combination of purified IgG and IgM NAb reduced the appearance of tumors slightly and increased the survival of recipients. The reconstitution of the xid‐defect in anti‐tumor NAb provides more direct evidence to substantiate a role for NAb in the defense against early tumor development in vivo and establishes a model for the dissection of the in vivo. mechanisms of the NAb anti‐tumor activity.
The capacity of human malignant B‐lymphocytes to disseminate in scid mice is correlated with functional expression of the fibronectin receptor α5β1 (CD49E/CD29)
Authors: Blase Lutz , Daniel Peter T. , Koretz Karin , Schwartz‐Albiez Reinhard , Möller Peter
Published: February 2007

The α5β integrin (CD49e/CD29), a heterodimeric membrane protein, is the “classical” fibronectin receptor on many cell types. During B‐cell ontogeny, expression of the α5‐subunit is developmentally regulated. The αβ1 is decisive for migration on fibronectin substrate and very likely cooperates with other adhesion molecules in transvascular trafficking. To test whether α5β influences local growth vs. disseminative spread of neoplastic B‐cells in vivo, human B‐cell lines mimicking different maturational stages were transferred s.c. into severe combined immunodeficiency (SCID) mice and examined for α5β expression and for adherence on fibronectin substrate in vitro and ex vivo. All cell lines were locally tumorigenic. Dissemination was observed in all animals carrying Nalm‐6 tumors, in one animal with a BL 60 and in 2 mice carrying a Raji tumor. By contrast, Daudi, BJAB and U266 tumors did not disseminate. As evidenced by immunohistochemistry and flow cytometry, all lines and their tumors were to various extents β1‐positive but showed considerable differences in α5 expression. The functional surface expression of α5β1correlated with fibronectin adherence of the lines. Daudi expressed α5β1 in a non‐functional configuration which was rendered functional only upon applying high concentrations of Mg++ and Mn++. B‐cell lines functionally expressing α5β1 at high or moderate levels disseminated in SCID mice while α5‐negative lines and Daudi did not. These results support the conclusion drawn from an earlier in situ analysis of human B‐cell lymphomas/ leukemias that the α5β1 integrin contributes to the disseminative phenotype of malignant B cells.
Prognostic value of cytosolic thymidine kinase activity as a marker of proliferation in breast cancer
Authors: Romain Sylvie , Christensen Ib J. , Chinot Olivier , Balslev Ingegerd , Rose Carsten , Martin Pierre‐Marie , Thorpe Susan M.
Published: February 2007

Thymidine kinase (TK) is involved in DNA synthesis by the salvage pathway. In this study, thymidine kinase (TK) was determined in routinely prepared cytosols of primary tumors from 290 breast‐cancer patients. Enzyme activity was measured using a radioenzymatic method optimized for detection of the fetal isoenzyme. High levels of TK (≥ 126 mU/mg protein) were positively associated with histological grade in both pre/peri‐and post‐menopausal patients. In pre/peri‐menopausal patients, high concentrations of TK were also found more frequently in progesterone receptor (PgR)‐negative tumors than in PgR‐positive samples. In post‐menopausal patients, high levels of TK were associated with large tumor size, estrogen receptor (ER) negativity and PgR negativity. In univariate analysis, high levels of TK were strongly associated with shorter overall survival in both pre/peri‐ (p = 0.001) and post‐menopausal patients (p = 0.02). Pre/peri‐menopausal patients whose tumors had high levels of TK also had an increased risk of relapse (p = 0.001). In multivariate analysis (including treatment protocol, patient age, lymph‐node involvement, tumor size, histological grade, ER and PgR status), TK status was found to be an independent prognostic factor for recurrence‐free survival in pre/peri‐menopausal patients with a weight similar to that of PgR status. In post‐menopausal patients, TK was the only factor selected for overall survival. © 1995 Wiley‐Liss, Inc.
Fibronectin isoforms are differentially expressed in normal and adenomatous human anterior pituitaries
Authors: Farnoud M. Reza , Farhadian Farahnaz , Samuel Jane L. , Derome Patrick , Peillon Françoise , Li J. Yuan
Published: February 2007

The expression of fibronectin (FN) isoforms containing the extradomains A and B (ED‐A+ and ED‐B+ FNs) as well as a differentially O‐glycosylated oncofetal form of the protein (onf‐FN) was investigated in 6 normal human anterior pituitaries and 25 human pituitary adenomas. In normal tissue, immunohisto‐chemical experiments showed the presence of FN molecules lacking the extradomains A and B (ED‐A− and ED‐B− FNs) without onf‐FN immunoreactivity. These proteins were localized in the connective tissue compartment and especially in the vessel walls. Analysis of FN mRNA demonstrated an in situ synthesis of ED‐A− and ED‐B− FNs in the normal anterior pituitary. By contrast, in the adenomas, immunoreactivity for ED‐A+ FN was observed in all cases. ED‐B+ and onf‐FN immunoreactivities were observed in 14 and 8 adenomas, respectively, regardless of the type, grade or invasiveness of the adenomas. ED‐A+ FN mRNA was expressed in all adenomas studied, and ED‐B+ FN mRNA was present in ED‐B+ immunoreactive cases only. In pituitary adenomas, these 3 forms of FN were specifically associated with the endothelium and vascular smooth‐muscle cells. Our results demonstrate that the processes of remodelling of the connective tissue compartment that occur in adenoma angiogenesis are associated with pre‐ and post‐translational alterations of FN synthesis leading to the expression of ED‐A+, ED‐B+ and oncofetal FNs. © 1995 Wiley‐Liss, Inc.
Expression of protein kinase C isoenzymes in colorectal cancer tissue and their differential activation by different bile acids
Authors: Pongracz Judit , Clark Penny , Neoptolemos John P. , Lord Janet M.
Published: February 2007

Expression of protein kinase C (PKC) isoenzymes was determined in paired samples of normal mucosa and colorectal cancer tissue from 13 patients. Total PKC activity in cancer tissue was significantly decreased compared to that in normal mucosa. Western blotting, using PKC isoenzyme‐specific antibodies, showed that two PKC isoenzymes, PKC β and PKC ϵ, were significantly decreased in cancer tissue. The level of PKC δ was increased in cancer tissue and the expression of PKC α and ζ was not altered significantly. Primary bile acids—cholic acid (CA) and chenodeoxycholic acid (CDCA)—and the principal secondary bile acids—deoxycholic acid (DCA), lithocholic acid (LCA) and ursodeoxycholic acid (UDCA)—were found to be potent and selective activators of partially purified PKC isoenzymes. PKC β I was the isoenzyme most effectively activated by secondary bile acids. Our data provide a model for the involvement of secondary bile acids in colorectal carcinogenesis through specific PKC isoenzyme modulation in colorectal mucosa. © 1995 Wiley‐Liss, Inc.
Transfected mutant p53 gene increases X‐ray‐induced cell killing and mutation in human fibroblasts immortalized with 4‐nitroquinoline 1‐oxide but does not induce neoplastic transformation of the cells
Authors: Kawashima Kunihiro , Mihara Koichiro , Usuki Hisashi , Shimizu Nobuyoshi , Namba Masayoshi
Published: February 2007

We introduced the mutant p53 gene (codon 273Arg‐His) into human fibroblasts (SUSM‐I cells) previously immortalized with 4‐nitroquinoline I‐oxide (4NQO) and obtained 2 clonal cell lines (SUSM‐I/p53‐I and SUSM‐I /p53‐6) expressing the mutant p53. Since the genetic background of SUSM‐I /p53 is the same as that of SUSM‐I except for the presence of the mutant p53, we expected to obtain more information on the mechanisms of p53 functions without the influence of other genetic differences by comparing cellular characteristics of both cell lines. SUSM‐I /p53 cells became about twice as sensitive to the cytotoxic effects of X‐rays as their parent SUSM‐I cells. Mutation frequency was determined by the appearance of hypoxanthine guanine phosphoribosyl transferase deficient (6‐thioguanine resistant) cells. As a result, the mutation frequency of SUSM‐I /p53 cells was about 5 times that of SUSM‐I cells transfected with or without the vector plasmid alone. Furthermore, when the SUSM‐I/p53 cells were exposed to X‐rays, the mutation frequency increased to about twice that of the non‐irradiated SUSM‐I /p53 cells. However, SUSM‐I /p53 cells showed neither anchorage‐independent growth in soft agar nor tumorigenicity in nude mice. These results indicate that the mutant p53 gene itself, which generally works in a dominant‐negative way on cellular carcinogenesis, is not sufficient for neoplastic transformation of immortalized human cells, and that additional genetic change(s) may be necessary for transformation. © 1995 Wiley‐Liss, Inc.
Regulation of lewis lung carcinoma invasion and metastasis by protein kinase A
Authors: Young M. Rita I. , Montpetit Michelle , Lozano Yvonne , Djordjevic Andelka , Devata Sandeep , Matthews John P. , Yedavallli Sreedhar , Chejfec Gregorio
Published: February 2007

Metastatic Lewis lung carcinoma (LLC‐LN7) cells have increased protein kinase A (PKA) activity and are more invasive in vitro than are non‐metastatic (LLC‐C8) cells. To determine whether PKA mediates the in vitro invasiveness and in vivo metastatic capabilities of these tumor cells, the LLC variants were stably transfected to over‐express the Cα subunit of PKA, and thus to have increased PKA activity, or to express a mutant cAMP‐resistant PKA Rlα subunit which blocks PKA activation. Wild‐type LLC‐LN7 tumor cells were invasive in vitro and in vivo, recurred after tumor excision and metastasized to the lungs. However, they lost these properties after transfection to express the mutant Rlα that blocks PKA activation. The non‐invasive, non‐recurring and non‐metastatic LLC‐C8 cells gained the capacity to invade, to recur following tumor excision and to metastasize when transfected to express the PKA Cα subunit. © 1995 Wiley‐Liss, Inc.
International patterns in the occurrence of Hodgkin's disease in children and young adult males
Authors: Macfarlane Gary J. , Evstifeeva Tatiana , Boyle Peter , Grufferman Seymour
Published: February 2007

It was reported over 20 years ago that there were distinct age‐specific patterns of Hodgkin's disease incidence in countries with different levels of economic development, and that there was an inverse relationship between the incidence of Hodgkin's disease in children and young adults within countries. Such observations were important, leading to hypotheses on the possibly infectious aetiology of the disease. Since the initial report, diverging trends in the incidence of Hodgkin's disease in children and young adults have been observed, and data from a much larger number of countries and cancer registries have become available. This led us to reassess international age‐related incidence patterns of Hodgkin's disease occurrence. Recent data show distinct differences in age‐specific Hodgkin's disease incidence patterns in different geographic regions. In general, the United States (US) and European countries had the pattern of low childhood rates and high young adulthood rates. However, countries which are not part of the European Union (EU), mainly Baltic states and countries of central and eastern Europe, showed a variant of this pattern: similarly high young adult rates, but rates in children higher than those in the US and EU. Incidence‐rate patterns for Latin American countries differed from those previously observed, with a shift towards patterns observed in more economically developed countries. Analysis of incidence data from earlier sources dating back to 1963 confirmed the original finding of an inverse association in incidence rates (c. 1963‐1967) using a selected group of cancer registries, but not when all data were considered. This association has become weaker over the past 20 years. Using current incidence rates (1983‐1987), no association between Hodgkin's disease rates in children aged 5 to 14 years (as well as 0 to 9 years) and young adults (20 to 34 years) was found. © 1995 Wiley‐Liss, Inc.
Abortion and the risk of breast cancer: A case‐control study in greece
Authors: Lipworth Loren , Katsouyanni Klea , Ekbom Anders , Michels Karin B. , Trichopoulos Dimitrios
Published: February 2007

We have examined the association between induced or spontaneous abortion and breast cancer risk in Greece. In a hospital‐based case‐control study in Athens, 820 patients with confirmed breast cancer were compared with 795 orthopedic patient controls and 753 healthy visitor controls, matched to cases by age and interviewer. Logistic regression was used to analyze the data, controlling for demographic, reproductive and nutritional variables. Odds ratio (OR) patterns were similar for the 2 control series, which were therefore combined to increase precision of the estimates. The risk for breast cancer was not increased for women who had a history of abortion, compared to nulliparous women with no history of abortion. Thus ORs and 95% confidence intervals were for nulliparous women with spontaneous abortion, 1.17 (0.64–2.13); for nulliparous women with induced abortion, 0.98 (0.56–1.73); for parous women with no abortion, 0.56 (0.31–1.01); for parous women with spontaneous abortion, 0.61 (0.33–1.14) and for parous women with induced abortion, 0.99 (0.56–1.74). When the analysis was restricted to parous women, using parous women with no history of abortion as the baseline, ORs and 95% confidence intervals were for induced abortion before first full‐term pregnancy, 2.06 (1.45–2.90); for induced abortion after first full‐term pregnancy, 1.59 (1.24–2.04) and for spontaneous abortion, 1.10 (0.82–1.40). Our findings suggest that an interrupted pregnancy does not impart the long‐term protective effect of a full‐term pregnancy attributable to terminal differentiation. © 1995 Wiley‐Liss, Inc.
In vivo growth inhibition of leukemia by 4‐amidinoindan‐1‐one 2′‐amidinohydrazone (CGP 48664a), a new inhibitor of S‐adenosylmethionine decarboxylase
Authors: Dorhout Bernard , Velde Roelf Jakob Te , Kingma Harri Ferwerdaanneke W. , De Hoog Elly , Muskiet Frits A. J.
Published: February 2007

We studied the in vivo growth‐inhibitory effect of the new S‐adenosylmethionine decarboxylase inhibitor 4‐amidinoindan‐I‐one 2′‐amidinohydrazone (CGP 48664A). L1210‐bearing DBA‐2 mice were treated with increasing CGP 48664A doses from 1 day after i.p. L1210 cell inoculation. Treatment was continued for 4 days, after which all mice were killed. CGP 48664A caused dose‐related exponential decreases of L1210 cell numbers and spermidine and spermine contents. Putrescine contents increased exponentially. Polyamine changes in spleen and liver were less profound. L1210 growth inhibition was not accompanied by changes in cell cycle phase distribution. It is concluded that CGP 48664A is an effective inhibitor of S‐adenosylmethionine decarboxylase but that CGP 48664A‐induced changes in intracellular polyamine compositions are not necessarily the cause of growth inhibition. © 1995 Wiley‐Liss, Inc.
Suppression of matrilysin inhibits colon cancer cell invasion in vitro
Authors: Yamamoto Hiroyuki , Itoh Fumio , Hinod Yuji , Imai Kohzoh
Published: February 2007

Matrilysin is a member of the matrix metalloproteinase gene family, which is believed to play an important role in tumor invasion and metastasis. We examined the effects of over‐ and under‐expression of matrilysin on the ability of colon cancer cells to migrate across an artificial membrane in vitro. Introduction of matrilysin caused colon cancer cells to become more invasive as assessed by an in vitro invasion assay. In contrast, expression of matrilysin was down‐regulated by all trans‐retinoic acid or by introduction of anti‐sense matrilysin in BM314 colon cancer cells. This down‐regulation caused these cells to become less invasive. We demonstrated a correlation between matrilysin level and the invasive potential of human colon cancer cells, implying an important role for matrilysin in the control of tumor invasion in vitro. © 1995 Wiley‐Liss, Inc.
A stress‐inducible 72‐kDa heat‐shock protein (HSP72) is expressed on the surface of human tumor cells, but not on normal cells
Authors: Multhoff Gabriele , Botzler Claus , Wiesnet Marion , Müller Eva , Meier Thomas , Wilmanns Wolfgang , Issels Rolf D.
Published: February 2007

It is suggested that members of the heat‐shock protein (HSP) 70 and 90 families are involved in intracellular antigen processing and the presentation of cell‐membrane‐anchored antigens. We show that non‐lethal heat shock (41.8°C) causes comparable rates of HSP72 (about 20x) and HSP73 (about 3x) synthesis in both tumor (including human Ewing's sarcoma, ES and osteosarcoma cells, HOS58) and normal cells (including EBV‐transformed B‐LCL, PBL and fibroblasts derived from healthy human volunteers). However, following non‐lethal heat stress and a recovery period at 37°C, flow cytometric analysis with a specific MAb showed HSP72 to be expressed only on the cell surface of tumor cells. The cell‐surface localization of HSP72 was confirmed by Western‐blot analysis of separated membranes and by immunoprecipitation with the HSP72‐specific MAb. In addition, co‐incubation of untreated tumor cells with supernatants from lethally heat‐shocked cells, which contain HSP72, did not lead to HSP72 cell‐surface expression. Thus, non‐specific association of HSP72 molecules with the outer plasma membrane is unlikely. In conclusion, despite comparable cytoplasmic HSP72 induction, human tumor cells differ from normal cells in their capacity to express HSP72 on their surface. This might imply clinical application as a means to target a stress‐inducible, tumor‐specific immune response. © 1995 Wiley‐Liss, Inc.
The presence of persistent high‐risk hpv genotypes in dysplastic cervical lesions is associated with progressive disease: Natural history up to 36 months
Authors: Remmink Ans J. , Walboomers Jan M. M. , Helmerhorst Theo J. M. , Voorhorst Feja J. , Rozendaal Lawrence , Risse Ellen K. J. , Meijer Chris J. L. M. , Kenemans Peter
Published: February 2007

To evaluate the clinical significance of HPV genotyping for the prediction of progressive cervical intraepithelial neoplasia (CIN) in women with cytomorphologically abnormal smears, a prospective, blind, non‐intervention study was performed. A total of 342 patients screened with cytomorphologically abnormal cervical smears were monitored every 3–4 months by cervical cytology, colposcopy and HPV testing using PCR. Women with progressive CIN disease were defined as patients developing lesions with a colposcopic impression of CIN III over more than 2 quadrants or resulting in a cytological smear equivalent to Pap 5. These patients were subsequently treated according to standard procedures. If any doubt arose about the true status of the patients (n = 75) these patients were censored and biop‐sied. The mean follow‐up time was 16.5 months (range 3–36 months). Nineteen women showed progressive CIN disease and all appeared to be continuously HPV‐positive from the start of the study. At biopsy, all these patients were histologically classified as CIN III. Seventeen of these women were positive for high‐risk HPV types. Two cases were classified as still unidentified HPV. No progression was seen in the absence of HPV DNA or in the presence of low‐risk HPV types. In life‐table analysis the cumulative rate of progressive, histologically verified CIN disease was 17% after 36 months. Further analyses showed that other risk factors such as age, sexarche, number of sexual partners or smoking hardly influenced the effect of HPV on progression. The results show that the continuous presence of high‐risk HPV types in women with cytomorphologically abnormal smears is a strong marker for progressive CIN disease. ©1995 Wiley Liss. Inc.
Human papillomavirus and cervical intraepithelial neoplasia grade II‐III: A population‐based case‐control study
Authors: Olsen Anne O. , Gjøen Kirsti , Sauer Torill , Ørstavik Ivar , Næss Oddvar , Kierulf Knut , Sponland Geir , Magnus Per
Published: February 2007

The association between certain human papillomaviruses (HPV) and cervical intraepithelial neoplasia (CIN) is well documented, but there is uncertainty about the strength of association and the role of co‐factors is unclear. This population‐based case‐control study in Norwegian women 20–44 years of age included 103 cases with histologically confirmed CIN II–III and 234 age‐matched and randomly selected controls. Cytological specimens from the cervix were analyzed using the polymerase chain reaction (PCR). In all, 91% of the cases and 15% of the controls were HPV DNA positive, giving a crude odds ratio (OR) of 67.2 (95% confidence interval: 28.6–157.5). The association between HPV 16 and CIN II–III was even stronger (crude OR = 123.9; 46.7 ‐ 328.5). In logistic regression analysis, additional to HPV, only a high number of sexual partners and a low educational level contributed independently to the risk. The adjusted OR for the association between HPV and CIN II–III was 72.8 (95% Cl: 27.6–191.9). The association between HPV and CIN remains very strong even after adjustment for proposed confounding factors. The results therefore support the role of HPV as a causative agent in the development of CIN. © 1995 Wiley‐Liss, Inc.
High frequency of EBV association with non‐random abnormalities of the chromosome region 1q21‐25 in aids‐related Burkitt's lymphoma‐derived cell lines
Authors: Polito Pietro , Cilia Anna M. , Gloghini Annunziata , Cozzi Marzia , Perin Tiziana , De Paoli Paolo , Gaidano Gianluca , Carbone Antonino
Published: February 2007

Chromosome lq abnormalities represent the second most frequent cytogenetic lesion of Burkitt lymphoma (BL) and acute lymphoblastic leukemia (ALL)‐L3. The most frequent change is partial duplication of the long arm of chromosome l, involving variable bands but consistently including 1q23. Among AIDS‐related BL similar chromosome 1q abnormalities have also been found. We have now characterized in detail the chromosome 1q abnormalities of 4 AIDS‐BL cell lines and compared them to other molecular features of the tumor clone, namely infection by Epstein Barr virus (EBV). Immunophenotypic characteristics were also assessed by conventional in situ immunocytochemical and flow cytometric procedures. The B‐cell origin of all cell lines was demonstrated by the expression of B‐cell‐restricted markers (e.g., CD 19). Analysis of 1g light chains confirmed their monoclonal nature. The t(8; 14) was present in 3 of the 4 lines, whereas variant translocation t(8;22) was detected in the remaining cell line. Additional chromosomal changes were found in all cases, with chromosome 1 being involved in all. Structural changes encompassed in each case the 1q21—25 bands, in either duplication or partial trisomy. EBER ISH studies identified EBV association in 3 of the 4 AIDS‐BL cell lines in contrast to previous studies of BL of immunocompetent individuals. Our findings of a high frequency of chromosome 1q abnormalities in EBV‐infected AIDS‐related BL cell lines demonstrate that such chromosomal abnormality and EBV positivity are not mutually exclusive and are possibly independent factors, whereas their close association in AIDS may be related to the immunodeficiency. © 1995 Wiley‐Liss, Inc.
CDK1 is a marker of proliferation in human lymphoid cells
Authors: Włowiec Dariusz , Deviller Philippe , Simonin Denis , Souchier Catherine , Rimokh Ruth , Benchaib Mehdi , Bryon Paul‐André , Ffrench Martine
Published: February 2007

To better understand the relationship between the proliferation of human lymphoid cells and the expression of cdk1, a catalytic subunit of the histone HI kinase (HIK), we examined its mRNA and protein content in 3 B‐cell lines: Ramos, Reh‐6 and IARC 963. Cells were elutriated according to their position in the cell cycle. Cell fractions were analyzed for cdk1 mRNA and protein cellular content by Northern blot and immunoblot, respectively, as well as for H1K activity. Both mRNA and protein amounts and H1K activity varied according to cell cycle phase, the lowest values being observed in G1‐enriched fractions. For comparison, elutriated fractions were also tested for the expression of cdk2 and cdk4 proteins. Both showed some variations among fractions, but they were less clear than those of cdk1. We also tested 29 samples of lymphoid neoplastic and non‐neoplastic tissues for proliferative activity (percentage of S and G2/M cells estimated by flow cytometry) and expression of cdk1, cdk2 and cdk4 proteins. We found a significant correlation between the percentage of cells in S or S + G2/M phases and cdk1 protein content but not cdk2 or cdk4 content. We conclude that cdk1 expression in human lymphoid cells varies during the cell cycle at both mRNA and protein levels. © 1995 Wiley‐Liss, Inc.
Modulation of growth and proliferation in squamous cell carcinoma by retinoic acid: A rationale for combination therapy with chemotherapeutic agents
Authors: Sacks Peter G. , Harris Daniel , Chou Ting‐Chao
Published: February 2007

We have previously shown that β‐all trans retinoic acid (RA) inhibits macrocellular growth of a multicellular tumor spheroid model for squamous carcinoma, as measured by spheroid size, but allows for continuing DNA synthesis and cell cycle progression, the two being reconciled by a cell death effect. DNA synthesis in the presence of growth inhibition suggested a rationale for examining combination chemotherapy with RA‐inhibited cells. To this aim, we have extended this observation to a series of 8 squamous carcinoma cell lines. Cells were treated with 1 μM RA for 7 days and cell growth parameters monitored. Although growth inhibition ranged from 0% (A431) to approx. 80% (MDA 886Ln), [3H]‐thymidine incorporation (cpm/μg protein) and percent S‐phase (by flow cytometry) in 7‐day RA‐treated cells was equal or higher than in their control vehicle‐treated cells in 7/8 SCC cell lines. Thus RA‐induced growth inhibition is not just cytostasis. Combination therapy was examined with MDA 886Ln, MDA 686Ln, 1483 and A431 cells pre‐treated for 7 days with 1 μM RA followed by cisplatin or 5‐fluorouracil treatment. An increased effectiveness for the combination was shown using 5‐day tetrazolium dye (MTT) growth assays when cells were growth‐inhibited by RA. Computerized analysis of data using median‐effect and isobologram techniques indicated that the interaction of RA with these chemotherapeutic agents was synergistic. With squamous carcinoma, RA treatment inhibits growth while allowing for continuing DNA synthesis, and these RA‐treated, growth‐inhibited cells exhibit increased sensitivity to chemotherapeutic agents. © 1995 Wiley‐Liss, Inc.
Systemic effects of cytokines released by gene‐transduced tumor cells: Marked hyperplasia induced in small bowel by γ‐interferon transfectants through host lymphocytes
Authors: Lollini Pier‐Luigi , D'Errico Antonia , De Giovanni Carla , Landuzzi Lorena , Frabetti Flavia , Nicoletti Giordano , Cavallo Federica , Giovarelli Mirella , Grigioni Walter F. , Nanni Patrizia
Published: February 2007

Cells transduced with cytokine genes are currently used to enhance the anti‐tumor and immunomodulatory effects of these molecules in cancer therapy. The sustained release of cytokine thus obtained can perturb many homeostatk systems of the host. We have previously shown that the murine mammary adenocarcinoma TS/A transfected with the murine γ‐in‐terferon (IFN‐γ) gene stimulates a strong immune response that impairs tumor growth. Mice bearing tiny tumors have serum IFN‐γ levels constantly exceeding 100 IU/ml. Therefore, we asked which systemic effects can be triggered in mice by such transfectants. BALB/c mice bearing tumors produced by clone 16.6000 cells (which release 6,000 IU/ml of IFN‐γ in culture) were compared to normal mice and to mice with tumors produced by parent cells transfected with the neomycin resistance gene (NEO cells, no IFN‐γ release). Histological studies revealed a marked hyperplasia of small bowel tn mice bearing 16.6000 tumors; the villi and crypts of these mice were >1.5 times longer than those of normal mice and of mice bearing NEO tumors. In vivo administration of bromodeoxyuridine evidenced a 2.5–3 times increase in the proliferative score of the intestinal crypts of mice bearing 16.6000 tumors compared to control mice. No intestinal alterations were observed in nude mice bearing 16.6000 tumors. T lymphocytes thus appear to play a causal role in this phenomenon. © 1995 Wiley‐Liss, Inc.
Is the risk of cancer of the corpus uteri reduced by a recent pregnancy? A prospective study of 765,756 norwegian women
Authors: Albrektsen Grethe , Heuch Ivar , Tretli Steinar , Kvåle Gunnar
Published: February 2007

We have examined the relations between the incidence of cancer of the corpus uteri and pregnancies in a cohort of 765,756 Norwegian women, contributing a total of 9,307,118 person–years in the age interval 30–56 years. Incidence rate ratios (IRR) were calculated by Poisson regression analyses of person–years at risk. Separate analyses were carried out for the 2 main histological subtypes, endometrial carcinomas (554 cases) and sarcomas (112 cases). We observed a decrease in risk of endometrial carcinoma with an increasing number of fullterm pregnancies (p < 0.001). The reduction in risk associated with the first pregnancy was more pronounced than that observed for any subsequent pregnancy. The risk of endometrial carcinoma increased with increasing time since last birth (IRR = 1.20, 95% CI = 1.08–1.34 per 5‐year time interval). The reduction in risk among parous women compared to nulliparous women diminished with increasing time since last birth. For endometrial carcinoma, the decrease in risk with increasing age at first and last birth disappeared after adjustment for time since last birth. For sarcomas, however, the relation with age at births remained in analyses adjusted for time since birth, and time since birth seemed to be of minor importance as an independent risk factor. Our results support the hypothesis that the reduction in risk of endometrial carcinoma associated with a pregnancy is related to a mechanical shed of malignant or pre‐malignant cells at each delivery. © 1995 Wiley‐Liss, Inc.
A single chain Fv derived from a filamentous phage library has distinct tumour targeting advantages over one derived from a hybridoma
Authors: Verhaar Marlies J. , Chester Kerry A. , Keep Patricia A. , Robson Lynda , Pedley R. Barbara , Boden Joan A. , Hawkins Robert E. , Begent Richard H. J.
Published: February 2007

Single‐chain antibodies (scFvs) can be derived from a monoclonal antibody (MAb) or produced directly using filamentous phage technology, where antibodies with desired binding and purification characteristics can be readily selected from libraries. To test the hypothesis that the latter approach is more useful, we compared 2 anti‐carcinoembryonic antigen (CEA) scFvs produced by these 2 different approaches. Our study showed that, both in the purification process and in the biodistribution pattern, MFE‐23, produced by filamentous phage technology, gave favourable results compared to A5‐SC, which is derived from the A5B7 MAb. This indicates the value of the filamentous phage approach for obtaining tumour‐targeting scFvs. © 1995 Wiley‐Liss, Inc.
MCF‐7 human mammary adenocarcinoma cell death in vitro in response to hormone‐withdrawal and dna damage
Authors: Wilson James W. , Wakeling Alan E. , Morris Ian D. , Hickman John A. , Dive Caroline
Published: February 2007

Anti‐oestrogens exert a tumoristatic effect on estrogen‐receptor‐positive breast carcinomas in vivo. At a cellular level this may reflect inhibition of cell proliferation and/or cell death counterbalanced by continued proliferation of a cell subpopulation. We evaluated the MCF‐7 human mammary adenocarcinoma cell line as an in vitro model to study the effects of the novel oestrogen antagonist ICI 182,720 on cell population dynamics (cell gain vs. cell loss). After oestrogen‐withdrawal monolayer cell number declined over 10 days, accompanied by cell detachment. This decrease in viable cell number was elevated 2‐fold by ICI 182,780. Detached cells exhibited DNA fragments of 50 and 300 kbp, typical of apoptotic cells. How‐ever, internucleosomal cleavage to 180 bp integer fragments was not seen, and these detached cells exhibited a morphology which was not consistent with apoptosis. The remaining attached monolayer cells were morphologically viable (>99%) with regard to both nuclear morphology and plasma membrane integrity. There was no difference in cell cycle phase distribution between oestrogen‐withdrawn and ICI 182,780‐treated cells; both induced accumulation in Gr phase. MCF‐7 cells were also exposed to a variety of DNA damaging agents known to induce apoptosis in other cell types. We could demonstrate only limited induction of morphologically recognisable apoptosis in MCF‐7 cells treated with methyl methanesulphonate. Our results add to the controversy surrounding the ability of the MCF‐7 cell line to undergo apoptosis in vitro in response to anti‐oestrogen therapies. © 1995 Wiley‐Liss, Inc.
Human breast cancer cell lines resistant to pure anti‐estrogens are sensitive to tamoxifen treatment
Authors: Lykkesfeldt Anne E. , Larsen Søren S. , Briand Per
Published: February 2007

The pure steroidal anti‐estrogens ICI 164,384 and ICI 182,780 are very potent growth inhibitors of the estrogen receptor‐positive human breast cancer cell line MCF‐7. However, long‐term treatment of MCF‐7 cells with 10−7 M concentrations of these compounds results in selection of proliferating colonies of resistant cells. Our report describes 4 ICI 164,384‐ and 3 ICI 182,780‐resistant MCF‐7 sublines established after long‐term treatment. Resistant sublines are estrogen receptor‐positive, and all sublines have lost expression and estrogen inducibility of the progesterone receptor protein. Based on IC50 concentrations, all tested resistant sublines had a reduced sensitivity to pure anti‐estrogens on the order of 100‐ to 1000‐fold compared with parent MCF‐7 cells. All resistant cell lines have survived propagation for more than 15 subcultivations in the presence of 10−7 M pure anti‐estrogen. The MCF‐7/182R‐6 subline has been tested for stability of resistance and appeared to be stably resistant after 13 weeks of propagation without the selective pressure of ICI 182,780. Cell lines resistant to the ICI 182,780 compound are cross‐resistant to the ICI 164,384 compound and vice versa. However, the sublines resistant to pure anti‐estrogens are sensitive to tamoxifen. Our results show that although the pure steroidal anti‐estrogens are very potent growth inhibitors, they do not circumvent development of resistance. © 1995 Wiley‐Liss, Inc.
The urokinase inhibitor p‐aminobenzamidine inhibits growth of a human prostate tumor in scid mice
Authors: Billström Anita , Hartley‐Asp Beryl , Lecander Ingegerd , Batra Satish , Åstedt Birger
Published: February 2007

Malignant cells possess a high degree of proteolytic activity in which the plasminogen activator system plays an important role. An increased expression of urokinase type plasminogen activator (uPA) is of significance for degradation of the extracellular tumor matrix, facilitating invasiveness and growth. Inhibition of the active site of uPA makes it possible to evaluate the significance of uPA in tumor growth. We report here experiments on a uPA‐producing human prostate xenograft (DU 145) using a competitive inhibitor of uPA, p‐aminobenzamidine. In vitro experiments with DU 145 cells showed that p‐aminobenzamidine caused a dose‐dependent inhibition of uPA activity. DU 145 cells were inoculated s.c. in SCID mice and, once tumors were established, treatment with p‐aminobenzamidine added to drinking water was started and lasted for 23 days. Mice receiving 250 mg/kg/day of p‐aminobenzamidine showed a clear decrease in tumor‐growth rate compared to the nontreated mice, resulting in 64% lower final tumor weight. In addition, uPA‐antigen levels in the membrane fractions of DU 145 tumors from p‐aminobenzamidine‐treated mice were found to be decreased by 59%. We also show that p‐aminobenzamidine has an anti‐proliferative effect in cell culture at low cell number, correlating with a dose‐dependent decrease in uPA production. In conclusion, we show that a low‐molecular‐weight uPA‐inhibttor, p‐aminobenzamidine, has a growth‐inhibitory effect on a solid uPA‐producing tumor. © 1995 Wiley‐Liss, Inc.