bluuebunny/crossref_metadata_2025 huggingface dataset:
Proliferating cell nuclear antigen and p53 expression as prognostic factors in T1‐2MO prostatic adenocarcinoma
Authors: Vesalainen Satu L. B. , Lipponen Pertti K. , Talja Martti T. , Alhava Esko M. , Syrjánen Kari J.
Published: February 2007

The expression of proliferating cell nuclear antigen and p53 protein was analysed by immunocytochemical methods (PC10, CMI antisera) in 139 patients with T1‐2MO prostatic adenocarcinomas followed‐up for > 12 years. p53 protein was expressed in 21 (15%) tumours (15%), the fraction of positive nuclei being very low (mean SE, 1% ± 0.7%). Accumulation of p53 protein in epithelial cells was independent of tumour stage and Gleason score, and had no effect on prognosis. In 4 cases, p53 protein was expressed only in stromal cells. The fraction of PCNA‐positive nuclei (evaluable in 116 cases) was higher in T2 than in T1 tumours (p< 0.001); furthermore, high Gleason score was positively correlated with PCNA positivity (p< 0.001). A finding of over 5% of PCNA‐positive nuclei predicted progression in T and M categories and were a sign of poor outcome. The fraction of PCNA‐positive stromal‐cell nuclei was related to T‐category with a borderline significance (p= 0.06). In a multi‐variate analysis of the prognostic factors, independent predictors of survival included Gleason score (p< 0.001), fraction of PCNA‐positive nuclei (p= 0.013), observation before therapy (p= 0.05), and T‐category (p= 0.07) in that order of significance. The results suggest that overexpression of p53 protein is of marginal prognostic value in local prostatic adenocarcinomas, whereas direct measurement of cell proliferation by PCNA immunolabelling provides important prognostic information in T1‐2MO tumours, in addition to the Gleason score.
Ethanol and breast cancer: An association that may be both confounded and causal
Authors: Katsouyanni Klea , Trichopoulou Antonia , Stuver Sherri , Vassilaros Stamatis , Papadiamantis Yannis , Bournas Nicholas , Skarpou Nafsika , Mueller Nancy , Trichopoulos Dimitrios
Published: February 2007

We have examined the association of alcohol intake during various life periods, by beverage category, with breast cancer risk in Greece. A hospital‐based case‐control study was performed in Athens, Involving 820 women with breast cancer as well as 795 orthopedic patients and 753 healthy visitor controls‐Relative risk patterns were very similar with either control series, which were therefore combined to increase precision of the estimates. Drinkers of beer were at significantly elevated risk for breast cancer [odds ratio (OR), 1.34 (95% confidence interval, 1.05‐1.71)]. However, drinkers of other alcoholic beverages were not at increased risk. Among beer drinkers there was no dose‐response, and drinkers of both beer and other beverages had a lower OR compared to drinkers of beer only. By contrast, drinkers of 3 or more glasses of alcohol per day, mostly of spirits, were at elevated risk for breast cancer [OR for 3 ‐ < 4 glasses per day, 3.01 (1.14‐7.95); OR for 4 or more glasses per day, 3.79 (1.05‐13.71)]. Reported frequency of consumption was a stronger predictor of breast cancer risk than either duration‐weighted total consumption or consumption before the age of 30 years. There were no coherent patterns for interaction with menopausal status, obesity or use of menopausal estrogens. The association of very low levels of alcohol intake with breast cancer risk may be due to confounding, whereas drinking 3 or more glasses of alcoholic beverages daily appears to genuinely increase breast cancer risk, perhaps by acting as a late‐stage growth enhancing factor. However, the data also are compatible with a linear relationship that has no threshold.
Differential expression of the cell‐cell adhesion molecule E‐cadherin in ascites and solid human ovarian tumor cells
Authors: Veatch Andrea L. , Carson Linda F. , Ramakrishnan S.
Published: February 2007

Advanced ovarian cancers contain 2 distinct phenotypic populations: (a) free‐floating tumor cells in the ascitic fluid and (b) solid tumors. Ascites cells are derived from the solid tumors and spread throughout the peritoneum. Changes in cell‐cell and cell‐extracellular matrix interactions are thought to be responsible for the origin of ascites cells. Since E‐cadherin molecules play a crucial role in the cell–cell interactions in epithelial cells, we investigated the expression of E‐cadherin in those 2 phenotypic populations. Paired samples of ascites and solid tumors were obtained from patients. Both primary tumors and tumor cells isolated from an experimental model showed a marked decrease in E‐cadherin expression in the ascites cells compared to the respective solid tumors. Semi‐quantitative, reverse transcriptase‐polymerase chain reaction (RT‐PCR) was used to determine the steady‐state levels of E‐cadherin‐specific mRNA. Results indicate that the primary tumors had significantly lower levels of E‐cadherin transcript in ascites cells when compared to their solid tumor counterparts. Changes in E‐cadherin expression were also reflected in the invasion capacity of tumor cells in vitro. Ascites cells were 4‐fold more invasive then solid tumor cells, suggesting that ascites cells are a highly malignant phenotype.
Infection of rats with human T‐cell leukemia virus type‐I: Susceptibility of inbred strains, antibody response and provirus location
Authors: Ibrahim F. , Fiette L. , Gessain A. , Buisson N. , De‐thé G. , Bomford R.
Published: February 2007

The susceptibilities of different strains of inbred rats to infection with the human T‐cell leukemia virus (HTLV‐I) after inoculation of human HTLV‐I producer cell lines were compared. The Fisher F344 and Brown Norway strains developed the highest antibody response to HTLV‐I, while the Lewis and BB strains were low responders. Antibodies against the HTLV‐I gag proteins, and env gp21 but not env gp46, were detected in Western blots with sera from HTLV‐I‐infected Fischer F344 and Brown Norway rats. These sera were inactive in an in vitro syncytium‐formation inhibition test The HTLV‐I provirus was detected by polymerase chain reaction in all Fischer F344, and some Lewis and Brown Norway rats, but not in the BB, which lack CP8+ T lymphocytes. The most frequent locations of the HTLV‐I provirus in the Fischer F344, Lewis and Brown Norway rats at 12 weeks after infection were the peripheral blood mononuclear cells (PBMC) and spinal cord. In a second experiment in Brown Norway rats, the provirus was again detected in the PBMC of rats at 11 weeks, but not at 22 weeks, and among the other organs tested at 22 weeks the sympathetic nerve ganglia were positive. It is concluded that HTLV‐I infection occurs in adult rats, but is suppressed with time.
Blockade of the insulin‐like growth‐factor‐I receptor inhibits growth of human colorectal cancer cells: Evidence of a functional IGF‐II‐mediated autocrine loop
Authors: Lahm Harald , Amstad Paul , Wyniger Josiane , Yilmaz Aysim , Fischer Jürgen R. , Schreyer Magali , Givel Jean‐Claude
Published: February 2007

Insulin‐like growth factors (IGFs) are potent proliferation stimulators for numerous tumor cells and often function as autocrine growth factors. We have previously shown that exogenous IGF‐I and IGF‐II enhance proliferation of colorectal carcinoma cells. The biological signal of both factors is transmitted through the IGF‐I receptor (IGF‐I‐R). This receptor was expressed in 12/12 colorectal carcinoma cell lines tested. αIR3, a neutralizing monoclonal antibody (MAb) directed against the human IGF‐I‐R, inhibited proliferation in 7/12 lines (Caco‐2, HT‐29, LS411N, LSS13, LS1034, WiDr and SW620), as reflected by a reduction of MTT conversion (19 to 42%), a decrease in cell number (39 to 72%) and an increase in doubling time (up to 2‐fold). In addition, in 4 cell lines (Caco‐2. LSS13, LS1034, WiDr) αIR3 suppressed colony formation in methylcellulose (40 to 84%). Excess of exogenous IGF completely neutralized αIR3‐mediated inhibitory effects. Northern blot analysis revealed abundant expression of 2 IGF‐II transcripts of 5.0 and 4.3 kb in LS1034 cells. In addition, we observed that growth inhibition by αIR3 was correlated with a more differentiated phenotype. Our results suggest that growth of many colorectal carcinoma cell lines is regulated by autocrine IGF‐II‐mediated stimulation of the IGF‐I‐R.
Progressive B‐cell chronic lymphocytic leukaemia frequently exhibits aberrant p53 expression
Authors: Aguilar‐Santelises Miguel , Magnusson Kristinn P. , Wiman Klas G. , Mellstedt Håkan , Jondal Mikael
Published: February 2007

We have analysed the p53 status in non‐progressive and progressive chronic B‐cell leukaemia (B‐CLL) by ELISA, immunoprecipitation, FACS and cDNA sequencing in relation to in vitro proliferation in response to Staphylococcus aureus strain Cowan I (SAC) and IL‐2. In FACS, cells from progressive leukaemia were found to over‐express p53 with a conformation recognized by PAb240. In a PAb240‐based ELISA, 60% of progressive B‐CLL were positive. DNA sequencing of p53 exons 5 to 9 revealed a codon 179 His to Gln change in one of the ELISA‐positive, progressive B‐CLL but failed to reveal any mutations in 4 other ELISA‐positive, progressive B‐CLL Among progressive B‐CLL populations, 10/14 responded by proliferation in vitro to SAC/IL‐2. In non‐progressive cells, low levels of p53 were found by FACS, none was positive in the PAb240 ELISA and only one case showed a weak proliferative response to SAC/IL‐2. Low p53 expression was also seen in different types of normal B cells, resting and activated, and in EBV‐transformed B‐cell lines, in contrast to the high expression observed in Burkitt lymphoma cell lines with verified p53 mutations. We conclude that progressive B‐CLL is characterized by aberrant p53 expression which may be a significant prognostic factor.
Prevalence of serum antibodies against the p53 tumor suppressor gene protein in various cancers
Authors: Angelopoulou Katerina , Diamandis Eleftherios P. , Sutherland Donald J. A. , Kellen John A. , Bunting Peter S.
Published: February 2007

We have developed 2 new quantitative methods for measuring anti‐p53 antibodies in human serum. Using these methods we analyzed 1,392 sera from patients with various malignancies and 230 sera from individuals without malignancy. Highest prevalence of anti‐p53 antibodies was associated with ovarian and colon cancers (15%), followed by lung (8%) and breast (5%) cancers. Prevalence in other malignancies was lower (<4%). In hospitalized patients and apparently healthy individuals, prevalence was very low (<2 and 1% respectively). Extremely high antibody concentrations (> 105 U/L) were found in 5 ovarian, 2 breast, I lung and I colon cancers. Sequential analysis of 6 positive samples has shown that the p53 antibody test may have potential for patient monitoring. The p53 antibody‐positive sera from breast cancer patients were associated with tumors that were steroid hormone receptor‐negative (p < 0.002). We propose that the measurement of p53 antibodies is a relatively specific serological test for cancer, which can be performed with easily automatable and quantitative methodologies and may be further exploited for patient monitoring, prognosis, diagnosis and probably screening for selected cancers.
Human neurological cancer cells express interleukin‐4 (IL‐4) receptors which are targets for the toxic effects of IL4‐pseudomonas exotoxin chimeric protein
Authors: Puri Raj K. , Leland Pamela , Kreitman Robert J. , Pastan Ira
Published: February 2007

Glioblastoma, glioma or neuroblastoma cells were examined for the expression of IL‐4 receptors (IL‐4R) by flow cytometric analysis and 125I‐IL‐4 binding. These cancer cell lines expressed IL‐4R which were of high affinity (KD = 700 × 10−12 M) on glioblastoma cells. To investigate the function of these receptors and to target potent cytotoxic antitumor agents to human neurological cancers, we utilized IL4‐PE4E, which is composed of IL‐4 and mutant Pseudomonas exotoxin (IL4‐PE4E). This chimeric molecule was cytotoxic toward human glioblastoma, neuroblastoma and glioma tumor cells in a dose‐dependent manner. The cytotoxicity of IL4‐PE4E was specific, since it was neutralized by excess IL‐4, and by an anti‐IL‐4 monoclonal antibody in all types of brain tumor tested. IL2‐PE4E and IL6‐PE4E were not cytotoxic, nor was an IL4‐PE4E mutant lacking ADP‐ribosylating activity, indicating the IL4‐PE4E‐mediated cytotoxicity of the brain tumor cells required both IL‐4R binding and enzymatic toxin activity. These data indicate that human neurological cancer cells express IL‐4R which are targets for the cytotoxic effects of IL4‐toxin. In addition, our data also suggest that IL4‐PE4E should be studied further as a potential treatment for human neurological cancers.
Comparison of effects of growth factors and protein kinase C activators on cellular sensitivity to cis‐diamminedichloroplatinum(II)
Authors: Basu Alakananda , Evans Rhobert W.
Published: February 2007

The anti‐proliferative activity of the DNA‐interactive anti‐cancer agent cis‐diamminedichloroplatinum(11) (cDDP) can be modulated by intracellular signaling systems. We have investigated the effects of growth factors on the sensitivity of human cervical carcinoma (HeLa) cells to cDDP. A 24‐hr pretreatment of HeLa cells with 10 ng/ml epidermal growth factor (EGF) or transforming growth factor‐α increased the anti‐proliferatfve activity of cDDP by 2‐ to 4‐fold. A similar pretreatment of HeLa cells with EGF did not alter cellular sensitivity to doxorubicin or vincristine. A brief exposure (15 min) to growth factors was not sufficient for cDDP sensitization. EGF caused a modest and transient increase in cellular diacylglycerol, the endogenous activator of protein kinase C. Bryostatin I, a partial agonist of protein kinase C, antagonized phorbol ester‐mediated cDDP sensitization but had no effect on EGF‐mediated sensitization to cODP. Both EGF and phorbol 12,13‐dibutyrate (PDBu) enhanced the rate of [195mPt]cDDP uptake but had no effect on the rate of [195mPt]cDDP efflux in HeLa cells. Bryostatin I reversed the increase in [195mPt]cDDP content by PDBu but failed to block EGF‐induced increase in [195mPt]cDDP accumulation. Therefore, although the mechanism of tDDP sensitization by both EGF and phorbol ester appears to involve enhanced drug uptake, they may utilize distinct signal transduction pathways.
Dietary fat, olive oil intake and breast cancer risk
Authors: Martin‐Moreno Jose M. , Willett Walter C. , Gorgojo Lydia , Banegas Jose R. , Rodriguez‐Artalejo Fernando , Fernandez‐Rodriguez Juan C. , Maisonneuve Patrick , Boyle Peter
Published: February 2007

As part of a population‐based case‐control study on diet and breast cancer in Spain, the role of dietary fat and vegetable oils in breast cancer etiology was examined. A validated, semi‐quantitative food‐frequency questionnaire was completed by 762 women, 18‐75 years of age, with histologically confirmed, newly diagnosed breast cancer, and 988 randomly selected female controls. For each food item and nutrient, the study subjects were divided into quartiles according to intake levels, with the lowest quartile serving as the reference category. Adjustment for total energy intake and other potential confound‐ers was made using multiple logistic regression for all women as well as separately for pre‐ and post‐menopausal women. Neither total fat intake nor specific types of fat were significantly associated with breast cancer in pre‐ or post‐menopausal women. However, higher consumption of olive oil (rich in monounsaturated fat) was significantly related to a lower risk of breast cancer [for highest vs. lowest quartile of consumption, odds ratio (OR) = 0.66; 95% Cl, 0.46‐0.97] with a significant dose‐response trend. While these findings do not support a relation between total fat intake and breast cancer risk, they do provide evidence for an inverse association between olive oil (and suggest one between monounsaturated fat) and risk of breast cancer.
c‐myc oncogene amplification and cytometric DNA ploidy pattern as prognostic factors in musculoskeletal neoplasms
Authors: Barrios Carlos , Castresana Javier S. , Falkmer Ursula G. , Rosendahl Ingvar , Kreicbergs Andris
Published: February 2007

The relationship between c‐myc oncogene amplification in neoplastic cells as determined by means of Southern‐blot analysis, and their nuclear DNA content as assessed by combined flow and image cytometry, was investigated in fresh tumor specimens from 33 patients with musculoskeletal neoplasms. Amplification, without rearrangement of the c‐myc proto‐oncogene, was detected in 4 out of 7 bone sarcomas and in 6 out of 26 soft‐tissue sarcomas, but in none of 3 benign giant‐cell bone tumors. Among the 10 cases with c‐myc amplification, 2 were found to be cytometrically DNA diptoid, 2 DNA tetraploid, and 6 DNA aneuploid. Conversely, there were 10 tumors displaying extremely aneuploid DNA patterns without c‐myc oneogene amplification. Thus, there was no relationship between c‐myc amplification and DNA ploidy; neither did the percentage of S‐phase cells, as determined by means of image cytometry, correlate significantly with the occurrence of c‐myc amplification. A surprising sex‐bias was observed; all 6 cases of c‐myc‐amplified soft‐tissue sarcomas occurred in females, whereas none of the 11 males with such sarcomas showed this amplification. When the clinical follow‐up data of the patients were scrutinized, it was found that the DNA ploidy pattern of the neoplastic cell nuclei, in combination with the S‐phase values, as well as the occurrence of c‐myc amplification, yielded prognostic information, being statistically significant 2 years after the diagnosis.
PCR‐RFLP studies on chromosome 3p in formaldehyde‐fixed, paraffin‐embedded cervical cancer tissues
Authors: Karlsen Frank , Rabbitts Pamela H. , Sundresan Vasi , Hagmar Bjørn
Published: February 2007

Loss of heterozygosity (LOH) has been extensively studied on the short arm of chromosome 3, and functional proofs have been obtained defining a tumor‐suppressor locus at 3p21‐22. We examined 31 paraffin‐embedded cervical cancer samples for LOH, using 5 PCR‐primer pairs, located around 3p21. Allele loss was found in 19 out of the 27 informative samples (70%) while 13 out of 23 informative samples (56%) had LOH located at 3p2l‐22. More of the human papillomavirus (HPV)‐positive samples had LOH compared to the HPV‐negative samples, giving only a weak association between loss of allele and HPV integration. Modifications of the DNA in the formaldehyde‐fixed samples were detected, and further studies will be required to clarify how such artifacts may affect restriction fragment length polymorphism (RFLP) studies on fixed tissues.
Role of clinical criteria in the diagnosis of hereditary non‐polyposis colorectal cancer (HNPCC): Results of a multivariate analysis
Authors: Percesepe Antonio , Anti Marcello , Marra Giancarlo , Roncucci Luca , Pahor Marco , Ccco Claudio , Armelao Franco , Gasbarrini Giovanni , De Leon Maurizio Ponz
Published: February 2007

Hereditary non‐polyposis colorectal cancer (HNPCC) is an autosomal dominant disease characterized by early‐onset colorectal tumors, primarily in the right colon, that are frequently associated with other cancers. In the absence of a reliable biomarker, clinical criteria for diagnosing HNPCC have been proposed by the International Collaborative Group on HNPCC (ICG‐HNPCC). However, these criteria are often too restrictive for application in small families. We analyzed 6 clinical/ pathological features suggestive of genetic colon‐cancer risk in 970 colorectal‐cancer patients defined according to the ICG criteria as HNPCC (96) or non‐HNPCC (874). Logistic regression analysis was used to determine their relative potentials for predicting the diagnosis of HNPCC. The most predictive were then used to estimate HNPCC risk levels within the non‐HNPCC group. Multivariate analysis showed the following associations with HNPCC diagnosis: vertically transmitted cancer (“highly predictive”), eariy‐onset (<50 yrs) colorectal cancer, aggregation of tumors in the nuclear pedigree and proximal‐colon tumors (the last 3 considered “predictive”). Re‐evaluation of all families revealed that 76 non‐HNPCC families (8.9% of the whole series) satisfied our highly predictive vertical‐transmission criterion plus at least one of the other “predictive” criteria. The presence of 2 consecutive generations affected by colorectal cancer or early primaries seems to be a major risk indicator of hereditary colorectal cancer. Using this approach we identified a large group of families that require further evaluation, although they do not currently meet the ICG‐HNPCC criteria for HNPCC.
Loss of heterozygosity at the RB locus correlates with loss of RB protein in primary malignant neuro‐endocrine lung carcinomas
Authors: Gouyer Valérie , Gazzeri Sylvie , Brambilla Elisabeth , Bolon Isabelle , Moro Denis , Perron Pascal , Benabid Alim Louis , Brambilla Christian
Published: February 2007

RB protein expression and loss of heterozygosity (LOH) in the RB gene were studied in 77 primary lung carcinomas of all histological types. RB protein expression was studied by immunohistochemistry with 3 anti‐RB antibodies, and was found altered in 23/29 (79%) neuro‐endocrine (NE) carcinomas and in 18/48 (37%) non‐NE carcinomas. RB gene allele status was studied with 3 probes detecting RFLP in RB locus. Fifty‐five patients were informative, and loss of heterozygosity was detected in 29 (52%) of the corresponding tumors with 1 of the 3 probes used; 89% of the informative NE carcinomas, excluding careinoids, and only 13% of the non‐NE carcinomas exhibited LOH and loss of RB‐protein expression. LOH at the RB locus was strongly correlated with the absence of RB protein in malignant NE carcinomas, and this association was strongly correlated with the neuro‐endocrine phenotype. Inactivation of the RB protein in primary NE carcinomas, excluding carcinoids, therefore seems to imply in the majority of cases the mutation of one allele and loss of the remaining allele of the Rfl gene, leading to loss of RB‐protein expression. In contrast, RB‐protein expression was independent of allele status in non‐NE carcinomas and carcinoids.
Formation of 8‐hydroxy‐2′‐deoxyguanosine and 4‐hydroxy‐2‐nonenal‐modified proteins in human renal‐cell carcinoma
Authors: Okamoto Keisei , Toyokuni Shinya , Uchida Koji , Ogawa Osamu , Takenewa Jun , Kakehi Ynshiyuki , Kinoshita Hidefumi , Hattori‐Nakakuki Yukari , Hiai Hiroshi , Yoshida Osamu
Published: February 2007

To study the possible involvement of reactive oxygen species (ROS) in the tumor biology of human renal‐cell carcinoma (RCC), we analvzed 35 cases of RCC for 2 parameters of oxidative damage: 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG), a mutation‐prone DNA‐ base ‐modified product, was measured by means of high‐performance liquid chromatography (HPLC) with an electrochemical (EC) detector, and 4‐hydroxy‐2‐nonenal (HNE)‐modified proteins were measured with a poly‐clonal antibody against HNE‐modified proteins. A 54% higher content of 8‐OHdG was found in RCC than in the corresponding non‐tumorous kidney, suggesting that the DNA of RCC is more exposed to ROS than is the DNA of non‐tumorous kidneys. Immunohistochemistry for HNE‐modified proteins showed a distinct staining pattern of fine to coarse granularity in the cytoplasm of RCC (n = 15), implying that lipid peroxidation products are located in cytoplasmic organelles. These results suggest that RCC constitutionally elaborates more ROS than is produced by the non‐tumorous parts of kidneys. No correlation was found between clinical stage, histology, age or sex and the 2 parameters examined.
Biological heterogeneity of er‐positive breast cancers in the post‐menopausal population
Authors: Romain Sylvie , Chinot Olivier , Guirou Olivier , Soullière Marc , Martin Pierre‐Marie
Published: February 2007

In the natural history of post‐menopausal patients with primary breast cancer, high estrogen receptor levels (ER) have been associated with a poor recurrence‐free survival. The purpose of this study was to investigate whether there are any biological intratumoral characteristics to support this puzzling clinical observation. In a population of 542 post‐menopausal, primary‐breast‐cancer patients, 3 normal distributions fitted into the frequency distribution curve of the logarithmically transformed ER‐EIA values. The biological profiles of the low ER group, and of the intermediate and high ER groups identified in the ER‐positive population were compared. Parameters correlated with ER functional aspect (progesterone receptors and PS2), receptors of epidermal growth factor (EGFR), protease cathepsin D and tumor proliferation (deduced from thymidine kinase activity) were analyzed. As previously reported, the levels of progesterone receptors and PS2 increased significantly from the low to the high ER groups. The highest levels of cathepsin D and thymidine kinase which have been previously related to a poor prognosis in breast cancer were found in the low ER group, but high levels were, surprisingly, also found in the high ER group. This study indicates that the ER‐positive post‐menopausal population is biologically heterogeneous. The high levels of thymidine kinase found in the high ER group suggest that overexpression of ER may be associated with proliferation enhancement, partly explaining the poor spontaneous prognosis related to this subset.
Conditional expression of human TNF‐α: A system for inducible cytotoxicity
Authors: Sparmann Gisela , Walther Wolfgang , Günzburg Walter H. , Uckert Wolfgang , Salmons Brian
Published: February 2007

Tumor necrosis factor‐α (TNF‐α) is currently being used in clinical trials for cancer treatment, but toxic side effects, due to systemic administration and high doses, are observed. Inducible expression of TNF may permit selective killing of tumour cells in gene therapy protocols without need for prolonged and/or high‐level TNF expression. A conditional TNF expression vector has been constructed in which the coding sequences of human TNF have been placed under the transcriptional control of the glucocorticoid‐regulated murine mammary tumour virus long terminal repeat (MMTV‐LTR). Negligible levels of TNF expression, associated with no phenotypic alterations, are observed in cells transfected with MMTV‐TNF vectors in the absence of glucocorticoid. Expression levels could be stimulated by the addition of the synthetic glucocorticoid dexamethasone. Increasing expression levels of TNF were associated with enhanced cytotoxicity. Our results suggest the potential use of inducible TNF systems for the treatment of tumours in gene therapy protocols.
Altered growth and phenotype in clonal mycN transfectants of the sk‐n‐sh neuroblastoma cell line
Authors: Gross N. , Miescher G. , Beck D. , Favre S. , Beretta C.
Published: February 2007

We have attempted to distinguish in human neuroblastoma between the effects of mycN on differentiation and its potential to promote malignant progression. Others have observed outgrowth of autocrine cells with evidence of an advanced malignant phenotype in mycN‐transfected clonal cell line derived from the single‐copy mycN neuroblastoma, SK‐N‐SH. We have now transfected the parental cell line with the same mycN expression vector and selected 5 clones characterized by unique and stable chromosomal integration sites and variable exogenous copy numbers. mycN gene expression was variable in the different clones and correlated roughly with the copy number of transfected mycN genes. Clones with minimal levels of mycN gene expression had a neuroblastic phenotype and low numbers of surface HLA class‐1 molecules. Clones with high levels of mycN expression had a Schwann/glial‐like phenotype with higher surface HLA class‐1 display without imbalance of expression of specific loci and accelerated growth. Two such clones were capable of anchorage‐independent growth in the absence of serum, and acquired tumorigenic properties. Our results show that exogenous mycN expression can be associated with a differentiation of neuroblastoma cells along the Schwann/glial pathway and can induce accelerated and autonomous growth.
Pleiotropic drug resistance and survival advantage in leukemic cells with diminished apoptotic response
Authors: Frankfurt Oskar S. , Seckinger Daniel , Sugarbaker Everett V.
Published: February 2007

Cell line R9 generated by continuous exposure of MOLT‐4 cells to adriamycin was cross‐resistant to a variety of unrelated drugs. The following data indicate that diminished apoptotic response was the mechanism of acquired pleiotropic drug resistance: (i) apoptosis was a common mechanism of cell death for agents expressing cross‐resistance; (ii) induction of apoptosis by drugs, medium depletion and serum deprivation was decreased in R9 cells; (iii) DNA degradation in apoptotic cells was lower in resistant lines, probably reflecting, a modification of apoptotic pathway in resistant cells; (iv) inhibition of cell division and DNA synthesis by drugs was similar in sensitive and resistant cells. These data indicated a similar level of initial damage, as typical for resistance based on modified apoptotic response. There was no difference inbcl‐2 protein level between sensitive and resistant cells. Thus acquired pleiotropic resistance and diminished apoptotic response in R9 cells were induced by a bcl‐2‐independent mechanism. Surface T‐cell antigen CD4 was expressed in MOLT‐4 and lost in R9 cells. The role of CD4 down‐regulation in apoptosis‐related drug resistance remains to be explored. The association between acquired pleiotropic drug resistance and increased survival capacity in unfavorable growth conditions indicated that drug‐induced selection of cells with diminished apoptotic response may stimulate neoplastic progression. Alkylating agents induced similar cytotoxicity and only slightly lower apoptosis in R9 cells in comparison with MOLT‐4 cells. Our data show that some drugs may overcome acquired pleiotropic drug resistance based on the modified apoptotic pathway.
Expression of interleukin 2 receptors on human carcinoma cell lines and tumor growth inhibition by interleukin 2
Authors: Yasumura Satoshi , Lin Wen‐Chang , Weidmann Eckhart , Hebda Patricia , Whiteside Theresa L.
Published: February 2007

We have previously shown that human squamous cell carcinomas (SCC) express the interleukin 2 receptor (IL2R)‐α and ‐β chains, and that the ligand, IL2, directly inhibits growth of the tumor in vitro and in vivo in the tumor xenograft‐nude mice model. We now show that the α and β chains of IL2R are expressed on a variety of human carcinoma cell lines and on normal human keratinocytes in early‐stage cultures. While all carcinoma cells in a population expressed IL2R‐α and ‐β proteins, in keratinocytes obtained from different normal donors, variable proportions of cells were positive, as measured by flow cytometry. The carcinoma lines and 2/5 keratinocyte lines studied were also found to contain transcripts for the IL2R‐γ chain detectable by combined reverse transcription‐PCR (RT‐PCR) and hybridization with the specific cDNA probe. Incubation of the gastric (HR) or renal cell carcinoma (RCC) cell lines, but not of other IL2R+ carcinoma cell lines or normal keratinocytes, in the presence of IL2 resulted in dose‐dependent inhibition of tumor cell growth. Monoclonal antibodies (MAbs) specific for IL2R‐β chain completely reversed this growth inhibitory effect of IL2. The ligand, IL2, also down‐regulated surface expression of its own receptor and of intercellular adhesion molecule‐I (ICAM‐I) or class I major histocompatibility complex (MHC) antigens on IL2R+ tumor cells. All carcinoma cells studied incubated in the presence of IL2 exhibited significantly increased sensitivity to growth‐inhibitory effects of other cytokines such as interferon (IFN)‐γ, tumor necrosis factor (TNF)‐α or transforming growth factor (TGF)‐β. IL2 inhibited growth of the HR cells by arresting a significant proportion of tumor cells in the Go/G1 phase of the cell cycle. Thus, IL2 can have direct effects on IL2R+ carcinoma cells, leading to changes in growth or to increases in sensitivity of tumor cells to cytostatic activities of other cytokines.
Correlation of cellular differentiation in human colorectal carcinoma and adenoma cell lines with metabolite profiles determined by 1H magnetic resonance spectroscopy
Authors: MaCkinnon Wanda B. , Huschtscha Lily , Dent Kathy , Hancock Rebecca , Paraskeva Christos , Mountford Carolyn E.
Published: February 2007

The aim was to determine whether proton magnetic resonance spectroscopy (MRS) could grade human colorectal cells of differing malignant potential. A cell model of tumour development and progression comprising 2 non‐tumorigenic adenoma lines and 4 carcinoma lines of increasing tumorigenicity was chosen. A gradual reduction in cellular differentiation and an accumulation of genetic alterations from adenoma to carcinoma characterized the selected cell lines. One‐dimensional and 2‐dimensional MRS showed that reduced differentiation in the cell model correlated with an increase in the levels of lipid, metabolites, the glycosylation intermediate uridine diphospho‐N‐acetylglucosamine and cell‐surface fucosylation. Mutations involving the K‐ras, APC and DCC genes are present both in adenoma‐ and in carcinoma‐derived lines in this model, but the first evidence of an abnormality in the p53 gene was concomitant with the cells' ability to grow as a tumour in athymic nude mice. This genetic change coincided with the detection, by MRS, of UDP‐hexose (ribose moiety, 2D MRS cross peak between H2 at 4.38 ppm and HI at 5.99 ppm) and the appearance of an additional fucosyl resonance (cross peak between‐CH3 at 1.41 and H5 at 4.30 ppm) in the least tumori‐genic of the carcinoma cell lines. An increase in complexity of the fucosylation spectral pattern was observed with further cellular de‐differentiation and increased tumorigenicity. Collectively these data support the existence of an adenoma‐carcinoma sequence.
Selective interactions between mammary epithelial cells and fibroblasts in co‐culture
Authors: Adam Liana , Crepin Michel , Lelong Jean‐Claude , Spanakis Emmanuel , Israel Lucien
Published: February 2007

Paracrine interactions between breast‐cancer cells (MCF7) and stromal fibroblasts were studied in relation to the presence of steroid hormones, using co‐cultures in which the 2 populations were separated by a microporous membrane. Densities and DNA‐synthesis rates of the co‐existing populations were interrelated. Proliferation was, therefore, viewed as the cumulative result of several factors, some of which are non‐specific, e.g., are density‐dependent, and some are specifically related to the feeders' origin and/or to culture conditions. Specific effects were measured and evaluated by stepwise analysis of covariance. MCF7 stimulated proliferation of fibroblasts differentially. Malignant‐tumour fibroblasts were stimulated more than non‐pathological ones. The magnitude of these effects was dependent on the presence of steroids. A similar analytical method was used for evaluating differential stromal influences on 4 epithelial phenotypic characters commonly used as prognostic markers. The estrogen‐receptor, progesterone‐receptor, pS2 and cathepsine‐D phenotypes of MCF7, as well as their interrelations, were dependent on the origin of the fibroblasts, i.e., embryonic or adult, normal or tumoral.
Spatial distribution of mitosis, apoptosis and small blood vessels in malignant diffuse follicular‐center‐cell lymphomas: A nearest‐neighbor analysis
Authors: Kraft Rainer , Barbini Paolo , Leoncini Lorenzo , Del Vecchio Maria T. , Megha Tiziana , Minacci Chiara , Spina Donatella , Massai Maria R. , Tosi Piero , Laissue Jean A. , Cottier Hans
Published: February 2007

To better comprehend the relationships between cell birth and cell death in neoplastic disorders, the topography of these events needs to be considered. We applied a computerized nearest‐neighbor analysis to malignant, diffuse follicular‐center‐cell lymphomas in order to examine the spatial distribution of pyknotic (apoptotic) cells/bodies (A) and mitotic figures (M) in relation to capillaries (C) and venules (V). The results revealed a complex dispersion pattern, with significant aggregations of A and M, in addition to an even greater random distribution component. The greatest clustering displayed by A was around capillaries (A‐C, 42% reduction of object frequency between the first and second distance class), followed by A‐A (33%), A‐V (24%) and A‐M (12%). Additional values for mitotic figures were: M‐M (45%), M‐V (33%) and M‐C (22%). These findings may reflect the relative importance of inherent properties of the neoplastic and host factors, respectively, in the regulation of cell birth and cell death rates. © 1994 Wiley‐Liss, Inc.
Cancer mortality following X‐ray treatment for ankylosing spondylitis
Authors: Weiss Helen A. , Darby Sarah C. , Doll Richard
Published: February 2007

Mortality has been studied in 15,577 ankylosing spondylitis (AS) patients diagnosed between 1935 and 1957 in the UK, of whom 14,556 received X‐ray treatment. By January 1, 1992 over half of the cohort had died. Among the irradiated patients, cancer mortality was significantly greater than expected from the national rates for England and Wales, with a ratio of observed deaths to expected (relative risk, RR) of 1.30, and significant increases individually for leukaemia, non‐Hodgkin's lymphoma, multiple myeloma and cancers of the oesophagus, colon, pancreas, lung, bones, connective and soft tissue, prostate, bladder and kidney. Among the unirradiated patients, cancer mortality was lower than expected from national rates (RR = 0.79). Among irradiated patients, the RRs for leukaemia, lung cancer, and all other neoplasms all decreased significantly with increasing time since first treatment following an initial increase. By 35 years after first treatment, the radiation‐related excess for lung cancer had completely disappeared, while for other neoplasms the RR remained significantly raised, although at a lower level than in earlier periods. Most irradiated patients received several courses of treatment within a 5‐year period. Based on a 1 in 15 random sample, the mean total body dose received in this period was 2.64 Gy, with the heaviest dose to the vertebrae. A linear dose‐response model for all neoplasms except leukaemia gave an excess RR of 0.18 Gy−1 in the period 5–24.9 years after first treatment, which decreased significantly to 0.11 Gy−1 in the period more than 25 years after first treatment. There was no evidence that a linear‐quadratic model fitted the data better than a linear model. There were significant dose‐response relationships individually for cancers of the lung, oesophagus, colon, pancreas, prostate, bladder and kidney. © 1994 Wiley‐Liss, Inc.
Enhanced expression of tissue inhibitor of metalloproteinase‐2 (TIMP‐2) in the stroma of breast carcinomas correlates with tumor recurrence
Authors: Visscher Daniel W. , Höyhtyä Matti , Ottosen Susan K. , Liang Chi‐Ming , Sarkar Fazlul H. , Crissman John D. , Fridman Rafael
Published: February 2007

The 72‐kDa (MMP‐2, gelatinase A) and the 92‐kDa (MMP‐9, gelatinase B) matrix metalloproteinases have been associated with tumor cell invasion and metastasis. Immunohistological staining of MMP‐2 and MMP‐9, basal lamina collagen IV and TIMP‐2 were performed on frozen sections of 83 invasive breast carcinomas. MMP‐2 and MMP‐9 were associated with neoplastic cell plasma membrane in 72% of cases and exhibited inter‐tumoral variability of staining intensity. MMP‐2 and MMP‐9 staining was not correlated with presence of metastases at time of diagnosis or with disease outcome. TIMP‐2 was detected in the peri‐tumoral stroma and was present in 87% of cases. Residual benign breast tissue was negative for TIMP‐2 staining. Neoplasms with diffuse TIMP‐2 staining (24%) recurred significantly more frequently (75% recurred) than cases with focal (42% recurred) or absent (27% recurred) TIMP‐2. Presence of collagen IV was negatively correlated with gelatinase staining. We conclude that up‐regulation of MMP‐2 and MMP‐9 expression in breast tumor cells is reciprocally correlated to collagen IV staining. Clinical outcome, however, is more closely related to the presence of TIMP‐2 than the corresponding MMPs. Enhanced TIMP‐2 expression, therefore, may denote a stromal response to tumor invasion, indicative of aggressive behavior in a subset of breast carcinomas. © 1994 Wiley‐Liss, Inc.
Pre‐morbid body size and the prognosis of women with breast cancer
Authors: Jain Meera , Miller Anthony B.
Published: February 2007

Obesity has been associated with poor prognosis of breast cancer, but studies investigating indicators of adiposity, independent of height, such as skinfold thickness and weight/height ratio, have been limited. From a cohort of 89,835 women in the National Breast Screening Study, data on histologically confirmed, incident cases of invasive carcinoma of breast, diagnosed between January 1981 to June 1992, were analyzed. Body measurements were taken at enrollment and other risk‐factor information from a self‐administered questionnaire completed prior to diagnosis of the breast cancer. Mortality data were obtained by linkage to Statistics Canada, provincial cancer registries, and annual follow‐up of cases through physicians. Of the 5 measures of body size studied among cases, poor prognosis was associated with higher triceps skinfold thickness only. With every 5 mm increase in skinfold, the risk of dying increased approximately 12% (relative risk, 1.12; 95% confidence intervals, 1.01 to 1.24) in the 1033 cases examined and adjusted for age at diagnosis, number of positive axillary nodes involved, and body weight. Compared with women in the lowest quartile of skinfold thickness, women in the fourth quartile had a relative risk (RR) of 1.64 (95% CI, 0.99 to 2.73). None of the other measures—weight, height, body mass index or weight/height ratio—reached any statistical significance. Since the reliability and validity of skinfold measures used in our study were never established, and no data were available for changes in body size after diagnosis, the results of this study on skinfold thickness should be used only as indicative of a need for further studies. © 1994 Wiley‐Liss, Inc.
Proliferating cell nuclear antigen (PCNA) in high‐grade malignant fibrous histiocytoma: Prognostic value in 48 patients
Authors: Dreinhöfer Karsten E. , Åkerman Måns , Willén Helena , Anderson Christina , Gustafson Pelle , Rydholm Anders
Published: February 2007

Grading based on histopathologic features is used to predict survival in soft‐tissue sarcoma. However, variations in clinical behavior between tumors of the same grade motivate a search for additional factors that correlate with prognosis. Proliferating cell nuclear antigen (PCNA) is expressed in proliferating cells during the G1 S and G2‐phases. To evaluate a prognostic implication of PCNA, the tumors of 48 patients with malignant fibrous histiocytomas (13 grade III, 35 grade IV) with a minimum follow‐up of 2 years were immunohistochemically studied. We used PC 10, a monoclonal antibody (MAb) directed against PCNA, which allows cell proliferation in formalin‐fixed, paraffinembedded tumor tissue to be evaluated. We appllied a semiquantitative PCNA grading scheme to all stained nuclei of an entire slide. The 3‐year metastasis‐free survival rate was 0.87 for patients in grade A (low PLNA rate), and 0.14 for patients in grade C (high PLNA). Our findings show that immunohistochemical evaluation of cell kinetics in soft‐tissue sarcomas by PCNA might be a useful adjunct to conventional tumor grading. © 1994 Wiley‐Liss, Inc.
Potent stimulatory effect of interleukin‐1α on apolipoprotein D and gross cystic disease fluid protein‐15 expression in human breast‐cancer cells
Authors: Blais Yves , Sugimoto Keiko , Carriere Marie‐Claude , Haagensen Darrow E. , Labrie Fernand , Simard Jacques
Published: February 2007

To better understand the multiple hormonal control of the expression of apolipoprotein D (apo‐D) and gross cystic disease fluid protein‐15 (GCDFP‐15, also designated prolactin‐inducible protein), which are 2 major proteins found in benign breastdisease fluid, we investigated their regulation by interleukin‐Iα (IL‐Iα) in the presence or absence of steroid hormones in ZR‐75‐I human breast cancer cells. Exposure of these cells to IL‐Iα decreased basal cell proliferation by half and markedly reduced the mitogenic action of 17β‐estradiol (E2), the halfmaximal inhibitory effect being exerted at 1.5 pM. In parallel, IL‐Iα stimulated apo‐D and GCDFP‐15 secretion with a similar potency. The antiproliferative effect of IL‐Iα was additive to the inhibition of cell proliferation caused by dihydrotestosterone (DHT) or the glucocorticoid dexamethasone (DEX). In parallel, IL‐Iα‐induced stimulation of apo‐D and GCDFP‐15 secretion was additive to that exerted by DHT or DEX. The sensitivity of the apo‐D and GCDFP‐15 responses to the stimulatory action of DHT or DEX was not changed by the presence of IL‐Iα. IL‐Iα also increased apo‐D and GCDFP‐15 mRNA levels. The present findings demonstrate the potent stimulatory effect of IL‐Iα on basal as well as androgen‐ and glucocorticoid‐induced apo‐D and GCDFP‐15 expression. The present data strongly suggest that IL‐Iα and steroids may modulate the secretion of these 2 proteins through different transduction pathways. © 1994 Wiley‐Liss, Inc.
MAGE‐1 gene product is a cytoplasmic protein
Authors: Schultz‐Thater Elke , Juretic Antonio , Dellabona Paolo , Lüscher Urs , Siegrist Walter , Harder Felix , Heberer Michael , Zuber Markus , Spagnoli Giulio C.
Published: February 2007

MAGE‐I gene encodes a human melanoma antigen, recognized by syngeneic cytotoxic T lymphocytes (CTL). MAGE‐I transcripts are also detectable in breast cancers, in non‐smallcell lung carcinomas and in central nervous system tumors. In order to identify, in cellular preparations, the protein encompassing the antigenic peptide, we generated a panel of monoclonal antibodies (MAbs) against the MAGE‐I gene product by using, as immunogen, a full‐length recombinant preparation (rMAGE‐I), obtained through expression cloning of the relevant gene in E. coli. Four reagents were obtained recognizing both rMAGE‐I and the 46‐kDa native protein in cell lines expressing MAGE‐I mRNA. No positivity could be detected in MAGE‐I‐mRNA‐negative melanoma lines. No surface labelling of MAGE‐I‐positive cell lines could be observed. In contrast, on permeabilization of MZ2 melanoma cells, all 4 MAbs induced efficient staining, as detected by cytofluorography. Fluorescence microscopy shows that MAGE‐I gene product is a cytoplasmic protein clustered in paranuclear organelle‐like structures. Thus, MAGE‐I protein location closely resembles that of P9IA and PI98 murine‐tumor antigens. © 1994 Wiley‐Liss, Inc.