bluuebunny/crossref_metadata_2025 huggingface dataset:
Addition of leucovorin in modulation of 5‐fluorouracil with methotrexate: Potentiating or reversing effect?
Authors: Van Der Wilt Clasina L. , Braakhuis Boudewijn J. M. , Pinedo Herbert M. , De Jong Marjon , Smid Kees , Peters Godefridus J.
Published: February 2007

Potentiation of the anti‐tumor activity of 5‐fluorouracil (5FU) by methotrexate (MTX) and leucovorin (LV) requires careful scheduling to achieve the most favorable interactions between these 3 drugs. In vitro the triple combination of MTX/5FU/LV was tested at 4 time intervals and with 3 MTX concentrations in 3 cell lines. MTX/5FU/LV was never superior to both 5FU/LV and MTX/5FU, moreover LV reduced growth inhibition by MTX/5FU in 2 cell lines. MTX/5FU was associated with only moderately increased levels of the 5FU metabolite FdUMP, while LV/5FU reduced the free FdUMP levels. MTX/LV/5FU had a dual effect and FdUMP levels gave no clear indication regarding the anti‐proliferative effect that could be expected. In vivo the time‐dependent potentiation of MTX on the 5FU anti‐tumor activity was confirmed for the treatment of HNX‐I4C‐bearing mice. Addition of LV to the MTX/5FU combination failed to cause a gain in anti‐tumor efficacy. For the treatment of Colon‐26‐bearing mice, MTX/5FU was comparable to LV/5FU, but the triple combination of MTX/LV/5FU seemed to have a better anti‐tumor effect. However, the toxicity of the latter treatment was greater than that of LV/5FU. The data from both in vitro and in vivo experiments failed to support the addition of LV to MTX/5FU therapy, possibly due to a reversing effect of LV. © 1995 Wiley‐Liss. Inc.
Differential expression of the H‐ras mutated and normal alleles in rabbit dmba‐induced keratoacanthomas
Authors: Matesanz Fuencisla , Oliva Maria Rosario , Villamarin Alicia , Kamino Hideko , Pellicer Angel
Published: February 2007

Keratoacanthomas (KAs) are benign and self‐regressing tumors in which a high incidence of the mutated H‐ras oncogene has been observed both in humans and in experimental models. To determine the level of expression of the mutated H‐rasallele with respect to its normal counterpart in 7,12‐dimethylbenz(a)anthracene (DMBA)‐induced KAs in rabbit skin, we have utilized a quantitative technique based on reverse transcription polymerase chain reaction (RT‐PCR) and selective cleavage of the mutated molecules of the H‐ras gene. Analysis of 16 KAs showed that the mutated H‐ras transcripts were up to 3‐fold more abundant than the non‐mutated H‐ras transcript in the different tumors. This higher expression of the mutated allele appears to correlate with increased differentiation in the KAs and in turn may contribute to tumor regression. © 1995 Wiley‐Liss, Inc.
Selective cytokine gene expression in renal cell carcinoma tumor cells and tumor‐infiltrating lymphocytes
Authors: Wang Qiu , Redovan Christine , Tubbs Raymond , Olencki Thomas , Klein Eric , Kudoh Seiji , Finke James , Bukowski Ronald M.
Published: February 2007

The progression of tumors such as renal cell carcinoma (RCC), despite the presence of substantial lymphocytic infiltrates (TIL), suggests that the ability of the local immune response to control tumor growth is impaired. Cytokine gene expression was examined to further investigate the nature of this response. Initial studies were performed with frozen tumors using PCR‐assisted mRNA amplification with cytokine‐specific primers for interleukin 10 (IL‐10), interleukin 2 (IL‐2) and interferon‐gamma (IFN‐γ). IL‐2 mRNA was not detected, despite the presence of T cells as defined by the expression of CD3γ mRNA. In contrast, mRNA for IFN‐γ was expressed in 4/9 and for IL‐10 in 5/9 tumors. To confirm this, 5 fresh tumor specimens were examined, and PCR demonstrated that IL‐10 mRNA was detectable in 4/5 tumors from which RNA was isolated at the time of nephrectomy. In these experiments multiple cycles and dilutions were employed to semi‐quantitate the expression of IL‐10. To identify potential sources of this cytokine in the tumor bed, IL‐10 mRNA expression in freshly isolated lymphocytes and tumor cells, TIL lines, cultured RCC and established RCC lines was examined. Our studies demonstrate that within the tumor TIL may be one source of IL‐10. Lymphocyte‐enriched populations from 4/5 tumors expressed IL‐10 mRNA as did 4/6 freshly isolated tumor cell preparations. IL‐10 gene expression was not detected, however, in tumor cells after one passage in vitro in short‐term cultured RCC tumor cells (passages 2–5) or in established RCC tumor cell lines. Finally, 4/9 CD4+ and 2/5 CD8+ TIL lines expressed IL‐10 mRNA either constitutively or after stimulation with anti‐CD3 antibody. This finding was associated with IL‐10 production in vitro. Our studies demonstrate that IL‐10 mRNA is frequently present in RCC tumors and may originate from the tumor‐infiltrating mononuclear cell population. © 1995 Wiley‐Liss, Inc.
Natural killer (NK) cells as effectors of antibody‐dependent cytotoxicity with chimeric antibodies reactive with human squamous‐cell carcinomas of the head and neck
Authors: Sung Myung‐Whun , Yasumura Satoshi , Johnson Jonas T. , Van Dongen Guus A. M. S. , Whitesidi Theresa L.
Published: February 2007

In patients with cancer, antibody‐dependent cellular cytotoxicity (ADCC) may be used as a laboratory test or for enhancing immunotherapy with murine monoclonal or chimeric mouse/human anti‐tumor antibodies (mMAbs or cMAbs, respectively). We have established an ADCC assay with IgG1 cMAb SF‐25, using human squamous‐cell carcinoma of the head and neck (SCCHN) cell lines as targets. By flow cytometry, all SCCHN cell lines tested expressed the antigen recognized by cMAb SF‐25. Trypsinization of the cell monolayers facilitated binding of cMAb SF‐25 to the antigen on the cell surface of SCCHN targets. Using the PCI‐50 SCCHN cell line as a target coated with this cMAb at the optimal concentration of 1.0 μg/ml, normal human peripheral blood mononuclear cells (PBMC, n = 28) were found to mediate ADCC at a mean level of 283 ± 42 (SEM) lytic units (LU20/107 effector cells). Non‐adherent monocyte‐depleted PBMC and natural killer (NK) cells purified by negative selection mediated significantly higher levels of ADCC than unseparated PBMC against SCCHN targets. NK cells, defined as CD3‐CD56' cells, could be effectively armed by cMAb SF‐25, as confirmed by flow cytometry and ADCC assays. IL2‐activated armed NK cells mediated higher levels of ADCC than non‐armed NK cells. Binding of cMAb SF‐25 to NK cells and their ADCC were enhanced by pre‐incubation with polyethylene glycol. Arming of NK cells with chimeric antibodies should be considered in developing novel strategies for treatment of human SCCHN, especially in the adjuvant setting. © 1995 Wiley‐Liss, Inc.
Correlation between functional and molecular analysis of mdr1 P‐glycoprotein in human solid‐tumor xenografts
Authors: Broxterman H. J. , Feller N. , Kuiper C. M. , Boven E. , Versantvoort C. H. M. , Teerlink T. , Pinedo H. M. , Lankelma J.
Published: February 2007

The contribution of P‐glycoprotein (Pgp) to multidrug resistance in human solid tumors is generally estimated from bulk mRNA measurements or immunohistochemistry, while direct measurement of the effect of Pgp on intracellular drug concentrations has not been pursued. We investigated the feasibility and sensitivity of a method for probing Pgp‐mediated drug transport in cells isolated from solid tumors, using xenograft models. Human tumor xenografts (XG) were grown by s.c. injection of Pgp‐expressing cell lines 2780AD, BRO/mdrl and KB8‐5. Tumor uptake of doxorubicin (DOX) after administration of DOX to the mice was determined. XG from untreated mice were enzymatically dissociated. The effect of the Pgp modulator bepridil on steady‐state cellular daunorubicin (DNR) and vincristine (VCR) accumulation and chemosensitivity of these XG cells was compared with its effects in the cell lines (CL). mdrl mRNA and Pgp (by flow cytometry) were measured. Also, the dependence on intraceilular ATP concentration, [ATP]i, of the modulator effect was determined in intact KB8‐5 cells. The results showed that i.v. administration of DOX to the mice led to lower DOX levels in the Pgp‐expressing XG than in the “sensitive” XG, suggesting the presence of an in vivo functional Pgp in these XG tumor models. Dissociated, viable XG cells appeared to have ATP levels sufficient to sustain Pgp‐ATPase‐coupled drug transport. This was inferred from experiments using KB8‐ 5 CL, which showed half‐maximal inhibition of DNR transport at an [ATP]i of 1 to 2 mM. The effect of bepridii on DNR and VCR accumulation and chemosensitivity in the XG cells was in accordance with the XG expression of mdr1/Pgp, In KB8‐5 XG cells, Pgp function was hardly detectable, in accordance with decreased mdrl/Pgp expression in vivo. In conclusion, Pgp activity can be determined in freshly dissociated XG human tumor cells. The results obtained with the more necrotic KB8‐5 XG may represent some of the interpretation problems arising when low levels of Pgp expression occur within a heterogeneous cell population, such as may be expected in clinical human tumors. Also our results indicate that Pgp activity may be impaired in vivo at [ATP]i below 2 mM, which are realistic values for human solid tumors. © 1995 Wiley‐Liss, Inc.
Two newly established cell lines derived from the same colonic adenocarcinoma exhibit differences in EGF‐receptor ligand and adhesion molecule expression
Authors: Solic Nicola , Collins Jane E. , Richter Audrey , Holt Susan J. , Campbell Ian , Alexander Peter , Davies Donna E.
Published: February 2007

Two morphologically distinct cell lines, GP2d and GP5d, derived from the same adenocarcinoma of the colon, have been established and characterised. Both clones have the same genetic changes, consistent with the usual pattern of tumour progression in colon cancer. The cells also have an inverted duplication of bands 10q11 to 10q21, but Southern blot analysis failed to identify any translocations involving the ret protooncogene, which maps to this region. GP2d grew by spreading from the edges of microcolonies to form a confluent layer of cells. GP5d grew in discrete islands of cells forming multilayered colonies. These differing patterns of growth correlated with variation in expression or cellular distribution of α2‐integrin, desmoplakin and e‐cadherin. Only GP2d responded to exogenously added epidermal growth factor (EGF), transforming growth factor‐alpha (TGFα) or insulin with an increase in cell numbers, even though both cell lines possessed similar numbers of EGF receptors. Analysis of EGF receptor ligand expression showed that GP5d cells expressed relatively more TGFα mRNA than did GP2d; in contrast, amphiregulin mRNA, which was abundant in GP2d, was virtually undetectable in GP5d. Even though GP5d failed to exhibit a growth response to EGF, it underwent a marked epithelial‐mesenchymal transition when treated with EGF, indicating separation of growth and morphological responses to EGF. © 1995 Wiley‐Liss Inc.
Radiation‐induced apoptosis in human sarcoma and glioma cell lines
Authors: Staffer Norbert J. , Stuschke Martin , Sak Ali , Stüben Georg
Published: February 2007

Six human soft‐tissue sarcoma and 14 glioma cell lines, exhibiting considerable differences in radioresponsiveness and histological grade of differentiation of the parental tumour, were examined with respect to apoptosis development after irradiation with 60Co γ‐rays. After test doses of 6 and 25 Gy, significant changes characteristic of apoptosis occurring within 6 to 30 hr were exhibited by only 2 differentiated sarcoma cell lines, EL7 and ESS2. The characteristic intemucleosomal fragmentation of DNA was detected as early as 6 hr after exposure of subconfluent monolayer cultures to 6 Gy. It was limited to cells that had detached from the culture plate, whereas adherent cells showed random degradation of DNA, namely after higher doses (25Gy) or longer incubation times (30 hr). As assessed by fluorescence microscopy of unfixed cultures stained with Hoechst 33342 and propidium iodide, the proportion of cells showing apoptotic bodies in non‐irradiated controls was < 0.l% and 0.3% for EL7 and ESS2, respectively. The doseresponse relationship for apoptosis was determined at 9 hr post‐irradiation. After 2 Gy, the percentage of apoptotic cells was elevated to 3.4% in EL7 and 4.5% in ESS2 cultures. Saturation was obtained above 6 Gy, with 8.4% apoptosis in EL7 and 15% in ESS2 after 25 Gy. Taken together, rapid ionizing‐radiation‐induced apoptosis seems to be limited to a subgroup of sarcomas and is unlikely to occur in gliomas. © 1995 Wiley‐Liss Inc.
Microvessel density and vessel invasion in lymph‐node‐negative breast cancer: Effect on recurrence‐free survival
Authors: Obermair Andreas , Kurz Christine , Czerwenka Klaus , Thoma Martin , Kaider Alexandra , Wagner Theresa , Gitsch Gerald , Sevelda Paul
Published: February 2007

Microvessel density (MVD) and blood and lymphatic vessel invasion (BLVI) were investigated with regard to their influence on the disease ‐ free survival (DFS) in node ‐ negative breast cancer patients. Paraffin embedded microsections of 230 patients with T1,2 NO breast cancer were immunohistochemically stained for factor Vlll ‐ related antigen. Every cluster consisting of more than highlighted endothelial cells was considered a countable microvessel. MVD was counted in 4 fields of 0.25 mm2each. All MVD values are given as value for the sum of 4 fields of 0.25 mm2each, that is, I mm2. BLVI was considered positive, when at least one tumor cell could be identified in a stained lumen. Out of 230 patients, 49 experienced local or distant recurrence and had a mean MVD of 72.4/mm2, whereas 181 patients who lived without recurrent disease had a mean MVD of 45.3/mm2. BLVI was negative in 6.2% of the cases with and in 93.8% of the cases without recurrent disease. BLVI was positive in 59.4% of the cases without and 40.6% of the cases with recurrent disease. MVD and BLVI remained the only significant prognostic factors of DFS in the Cox ‐ Model. Tumor size, histological grade, and hormonal ‐ receptor status were not prognosticallv relevant in the Cox ‐ model. 10 ‐ year ‐ DFS was 93.3% in BLVI ‐ negative/MVD ⩽ 40/mm2patients, 88.1% when MVD was high or BLVI was positive and 48.9% in BLVI positive/MVD ⩽ 40/mm2patients. Our present data indicate that MVD and BLVI identify a very ‐ low risk group among node ‐ negative breast cancer patients, who will not benefit from systemic adjuvant therapy. MVD and BLVI should be used as stratification criteria in clinical trails on node ‐ negative breast cancer patients. © 1995 Wiley‐Liss Inc.
Purine enzyme profile in human colon‐carcinoma cell lines and differential sensitivity to deoxycoformycin and 2′‐deoxyadenosine in combination
Authors: Camici Marcella , Turriani Maura , Tozzi Maria Grazia , Turchi Gino , Cos Joan , Alemany Cristina , Miralles Angela , Noe Veronique , Ciudad Carlos J.
Published: February 2007

Different cell lines, 2 from human colon carcinoma (LoVo and HT29) and 1 from Chinese hamster ovary (CHO K‐1), were examined to assess the effect of deoxycoformycin (dCF), an inhibitor of adenosine deaminase (ADA), and 2′‐deoxyadeno‐sine (dAdo) on their growth. When used alone, neither dCF or dAdo were cytotoxic for the 3 cell lines, while their combination caused inhibition of cell growth, with the following sensitivity: CHO K‐1 > LoVo > HT29. We studied the pattern of enzymatic activities involved in the metabolism of dAdo in the 3 cell lines. The phosphorylation of dAdo by adenosine kinase appears to play a central role in the toxicity of the deoxynucleoside in combination with dCF. In fact, CHO K‐1 cells, which are the most sensitive, possess the highest level of this enzyme. Moreover, the cytotoxic effect was almost completely reversed in the 3 cell lines when inhibitors of adenosine kinase, such as 5′‐amino‐5′‐deoxyadenosine and iodotubercidine, were added to the culture medium together with dCF and dAdo. In addition, baby hamster kidney (BHK) adenosine‐kinase‐deficient (AK−) cells were highly resistant to this treatment. Uptake inhibition of dAdo using dipyridamole also caused reversal of the toxicity. The AMP and deoxyAMP dephosphorylating activities, much lower in the CHO K‐1 cells, also appear to play a central role in the toxicity of dAdo when adenosine deaminase is inhibited. However, our data suggest that other factors may modulate the toxic effect, such as S‐adenosyl‐homocysteine‐hydrolase inhibition by dAdo at high concentrations. © 1995 Wiley‐Liss, Inc.
Induction of 2,5 oas gene expression and activity is not sufficient for IFN‐γ‐induced neuroblastoma cell differentiation
Authors: Corrias Maria Valeria , Gribaudo Giorgio , Guarnaccia Francesco , Ponzoni Mirco
Published: February 2007

We showed earlier that interferon‐γ is a powerful inducer of differentiation of human neuroblastoma (NB) cells. Although 2′,5′oligo‐adenylate synthetase (2,5 OAS) may play a role in mediating the anti‐proliferative and/or differentiative effects of interferons (IFNs), direct evidence is lacking. We have investigated gene and protein expression of the 4 different 2,5 OAS isoforms and their cumulative enzymatic activity in a previously characterized IFN‐γ‐sensitive human NB cell line, LAN‐5. Analysis of total and poly(A)+ RNA by Northern Mot and RT‐PCR indicated that expression of the mRNA coding for the 40‐, 46‐and 69‐kDa Isoforms was induced in a time‐ and dose‐dependent manner, reaching a maximum after a 36‐hr treatment with 1000 IU/ml of IFN‐γ. In the absence of treatment, only the mRNA for the 69‐kDa isoform was detectable by RT‐PCR. Inhibition of transcription with actinomycin D showed that 2,5 OAS mRNA was quite stable, with a half‐life of about 4 hr. With respect to the protein content, no 2,5 OAS isoform was present in proliferating LAN‐5 cells; following IFN‐γ treatment, the 100‐, 69‐ and 46‐kDa isoforms became detectable. Accordingly, 2,5 OAS enzymatic activity, virtually undetectable in untreated LAN‐5 cells, increased up to 132 pmol oligoadenylate/μg protein/hr after 48 hr of treatment, then slowly decreased, remaining detectable up to 96 hr. However, the 2,5 OAS proteins required an exogenous activation by synthetic dsRNA to exert enzymatic activity. It is therefore conceivable that they do not play a biological role in NB cell functions. Moreover, an increase in 2,5 OAS enzymatic activity was also observed in NB cells resistant to the differentiation‐promoting activity of IFN‐γ, further suggesting that 2,5 OAS induction was not sufficient to trigger IFN‐γ‐dependent neuronal maturation. Furthermore, other differentiation‐inducing agents, such as retinoic acid and cytosine arabinoside, or complete proliferative arrest produced by serum deprivation, failed to enhance 2,5 OAS activity, thus indicating that the 2,5 OAS system is not directly involved in mediating other differentiative pathways of NB cells. © 1995 Wiley‐Liss, Inc.
The prognostic influence of neuroendocrine cells in prostate cancer: Results of a long‐term follow‐up study with patients treated by radical prostatectomy
Authors: Noordzij Marinus A. , Van Der Kwast Theodorus H. , Van Steenbrugge Gert Jan , Hop Wim J. C. , Schröder Fritz H.
Published: February 2007

The distribution of immunohistochemically defined neuroen‐docrine (NE) cells in benign, pre‐cancerous and neoplastic prostatic tissues and the prognostic value of these cells in prostate cancer were studied in the radical prostatectomy specimens of 90 patients from whom complete long‐term follow‐up data were available. The tissue blocks containing all the different Gleason patterns observed in a particular tumor were selected and immunostained. Since chromogranin B stained only a few cells compared to chromogranin A (CgA), NE cells were only defined by their reactivity with CgA. A semiquantificative CgA score was assessed for all distinct pathological areas. Cox's regression model was used to analyze the influence of final TNM classification (TNM, 1992), Gleason sum score (GSS), age and CgA score on the probability of progression and tumor‐specific death. NE cells were demonstrated in all normal prostatic tissues and in most hyperplastic and intra‐epithelial neoplastic lesions. CgA staining was seen in 78% of the tumors. CgA scores were not related with Gleason growth patterns, GSS or TNM classification and had no prognostic value. The independent prognostic variables in Cox's regression model were: GSS and pT stage for progression and GSS for tumor‐specific survival. Theoretically, NE cells could influence tumor behavior and this discrepancy suggests the need for experimental studies to investigate the role of NE cells in the normal and neoplastic prostate. © 1995 Wiley‐Liss Inc.
Effect of different platelet agonists on intracellular free CA++ concentrations in human tumor cells: Possible role in tumor growth
Authors: Saporiti Anna , Brocchieri Alessandra , Porta Camillo , Moroni Mauro , Grignani Guido
Published: February 2007

Modulation of cytoplasmic Ca++ concentration is a mechanism common to signal transduction pathways regulating many cellular phenomena, including the interactions of tumors with the hemostatic system. We have investigated the pro‐aggregating and pro‐coagulant activities of human tumor cell lines cultured in vitro and the ability of different platelet agonists to induce Ca++ transients in these cells. Cells of a malignant mesothelioma line activated platelets by a thrombin‐dependent mechanism; on the contrary, HeLa cells, derived from a uterine cervical cancer, possessed ADP‐dependent pro‐aggregating activity, and DND‐1A melanoma cells did not stimulate platelet aggregation. All cell lines showed a tissue‐factor‐like procoagulant property, more pronounced in mesothelioma cells. Furthermore, ADP was able to induce a transient increase in cytoplasmic Ca++ concentration in tumor cells from all lines; collagen showed this effect in mesothelioma cells and in HeLa cells, and thrombin was effective only in mesothelioma cells. PAF never induced Ca++ fluxes in any of the cell lines investigated. Finally, the calcium‐channel blocker verapamil inhibited agonist‐induced Ca++ transients in tumor cells and in vitro tumor‐cell growth. These data may help to identify new possible mechanisms of the 2‐way interaction of tumors with the hemostatic system. © 1995 Wiley‐Liss Inc.
Treatment of wistar rats with a renal carcinogen, ferric nitrilotriacetate, causes dna‐protein cross‐linking between thymine and tyrosine in their renal chromatin
Authors: Toyokuni Shinya , Mori Toshiaki , Hiai Hiroshi , Dizdaroglu Miral
Published: February 2007

Ferric nitrilotriacetate (Fe‐NTA) induces renal proximal tubular damage associated with lipid peroxidation and oxidative DNA base modifications that finally leads to a high incidence of renal adenocarcinoma in rodents. In the present study, we report on the in vivo formation of DNA‐protein cross‐links (DPCs) involving thymine and tyrosine in the renal chromatin of Wistar rats treated with single or repeated i.p. administration of Fe‐NTA. Analyses of chromatin samples by gas chromatography/mass spectrometry revealed a significant increase in the amount of 3‐[(1, 3‐dihydro‐2, 4‐dioxopyrimidin‐5‐yl)‐methyl]‐L‐tyrosine (Thy‐Tyr cross‐link) 24 and 48 hr after the administration of Fe‐NTA. At 19th day of Fe‐NTA treatment, the amount of Thy‐Tyr cross‐link decreased to the control level, indicating the presence of cellular repair activity. Thy‐Tyr cross‐link may play a role in the genetic alteration of this renal carcinogenesis model, since mitoses for regeneration of renal proximal tubules were closely associated with the increase in DPCs. © 1995 Wiley‐Liss Inc.
Manifestations of cancer cachexia induced by colon 26 adenocarcinoma are not fully ascribable to interleukin‐6
Authors: Soda Kuniyasu , Kawakami Masanobu , Kashii Akiyoshi , Miyata Michio
Published: February 2007

In order to further clarify the role of interleukin 6 (IL‐6) in the pathogenesis of cachexia, recombinant human IL‐6 (hIL‐6) was administered s.c. by osmotic pump for 9 days at a dose of 1 or 10 μg/day into CDFI mice inoculated with a non‐cachexia‐inducing subclone of colon 26 adenocarcinoma (clone 5), or with a cachexia‐inducing subclone (clone 20) of this malignancy. The serum level of IL‐6 in non‐cachectic mice with clone‐5 tumors was 35% lower than in cachectic mice bearing clone 20 of colon 26 adenocarcinoma on the 19th day after tumor inoculation. IL‐6 administration induced anemia, thrombocytosis and visceral organ hypertrophy not only in mice with clone‐5 tumors but also in control mice with no tumor burden. Lipolysis and proteolysis became conspicuous when a large dose (10 μg/day) of IL‐6 was infused into mice with clone‐5 tumors. However, IL‐6 supplementation did not induce loss of body weight, a decline in food intake or lymphocytopenia, which were characteristically observed in cachectic mice with clone‐20 tumors. In conclusion, IL‐6 appears to be a permissive factor for the development of cachexia but, while it can induce some of the symptoms typical of cachexia, it cannot in itself induce the full cachectic syndrome. © 1995 Wiley‐Liss Inc.
Translocation t(9;22)(q22;q12) is a primary cytogenetic abnormality in extraskeletal myxoid chondrosarcoma
Authors: Stenman Göran , Andersson Helene , Mandahl Nils , Meis‐Kindblom Jeanne M. , Kindblom Lars‐Gunnar
Published: February 2007

The cytogenetic and in vitro growth characteristics of 3 cases of extraskeletal myxoid chondrosarcoma (EMC) are described. In cell culture, the tumor cells retained the immunocytochemical and ultrastructural characteristics of EMC. Cytogenetically, 2 of the cases showed an apparently identical t(9;22)(q22;q12). In one case, the t(9;22) was found together with a dup(1)(q12q44), and in the other case it was found together with several other aberrations. The third case had an inv(10)(p11.2q22) as the sole karyotypic abnormality. Of 4 cases of EMC previously analyzed, 2 showed a t(9;22)(q22;q11–12) and one case a t(9;22;15)(q31;q12.2;q25). Thus, 5 out of 7 cases of EMC showed recombination between 9q22–31 and 22q11–12, indicating that this represents a tumor specific abnormality. The breakpoints on 22q were in all 5 cases Cytogenetically indistinguishable from those seen in Ewing's sarcoma with t(11;22), clear‐cell sarcoma with t(12;22), and desmoplastic small round cell tumors with t(11;22). Molecular cloning of these translocation breakpoints revealed involvement of the EWS gene (located at 22q12) in all 3 tumor types. These observations raise the intriguing question of whether the EWS gene might also be involved in the t(9;22) in EMC. © 1995 Wiley‐Liss, Inc.
Breast‐cancer risk and oral contraceptive use in slovenian women aged 25 to 54
Authors: Primic‐Žakelj Maja , Evstifeeva Tatiana , Ravnihar Božena , Boyle Peter
Published: February 2007

Results of a previous case‐control study in Slovenia showed a significantly elevated risk of breast cancer for ever‐OC users aged 25 to 54 years. A further study was conducted in 1988–1990 in the whole of Slovenia, employing more rigorous epidemiological methodology. Cases were 624 women with breast cancer, aged 25 to 54 years, diagnosed at the Institute of Oncology in Ljubljana and other Slovenian hospitals. Controls were 624 women identified through the Population Registry, randomly selected and matched with cases by date of birth and commune of residence. Data were collected by personal interview, using coloured photographs of packages of all OC on the Slovenian market since 1964. A calendar of reproductive life events was constructed with participants to improve estimation of exposure. The adjusted odds ratio (OR) for ever‐users was 1.09. There was no increase in risk with total duration of use, interval since first use, age at starting OC, according to use before or after first delivery and time between menarche and age at first use. Increased risk (OR = 2.92) was found for OC users at the time of diagnosis and for those stopping them less than 6 months before (current users). The risk was not increased for those who stopped OC more than 6 months before diagnosis. The results of this study are consistent with most studies showing no overall effect of OC in women aged till 55 years ever using them. Increased risk of breast cancer in current OC users suggests a possible promoting effect of the pill in susceptible women, and indicates the need for careful breast surveillance of these women while they are using OC and in the period immediately following cessation. © 1995 Wiley‐Liss, Inc.
Studies of in vivo recruitment and activation of cytotoxic lymphocytes using a gelatin‐sponge model of concomitant tumor immunity
Authors: Park Julie A. , Brown Rebecca A. , Kurt Robert A. , Akporiaye Emmanuel T.
Published: February 2007

We have earlier described a sponge model of concomitant tumor immunity that permits the capture and isolation of effector T lymphocytes that mediate the rejection of a secondary EMT6 tumor implant In this study, we have employed the sponge model to study lymphocyte homing and in situ activation during the development of concomitant tumor immunity. Our results demonstrate that EMT6‐specific CTL in animals bearing primary EMT6 tumors home preferentially to sponges implanted with EMT6 tumor cells, as compared with contralateral sponges lacking tumor cells or sponges injected with antigenically distinct 168 tumor cells. We further show that recruitment is selective and is not in response to a foreign‐body reaction to the sponge. In addition, we show that EMT6‐specific CTL were recovered from sponges injected either with intact EMT6 tumor cells or with a mixture of EMT6‐derived membranes and supernatant. In contrast, cells accumulating in sponges injected with membranes or supernatant alone were not cytolytic. Thus, maximal recruitment, retention, and activation of CTL precursors require putative chemo‐attractive factors secreted by tumor cells as well as interaction with tumor antigen. © 1995 Wiley‐Liss, Inc.
The epithelial/carcinoma antigen EGP‐1, recognized by monoclonal antibody RS7–3G11, is phosphorylated on serine 303
Authors: Basu Amartya , Goldenberg David M. , Stein Rhona
Published: February 2007

RS7–3G11 is a murine monoclonal antibody (MAb) raised against human non‐small‐cell lung carcinoma, and is under clinical evaluation. The epithelial/carcinoma antigen EGP‐1, defined by RS7–3G11, was isolated and purified to homogeneity from a cervical carcinoma cell line, ME 180. EGP‐I is a glycoprotein with an average molecular mass of 47.8 kDa. Metabolic labeling of the antigen with 32P‐orthophosphate and subsequent immunoprecipitation with RS7–3G11 showed that it is a phospho‐protein. Phosphoamino acid analysis of the in vivo phosphorylated EGP‐1 revealed that the phosphorylation is on serine. In vitro analysis with purified antigen demonstrated that protein kinase C, and not protein kinase A, is involved in phosphorylating the antigen in vitro. In vitro analysis indicated a stoichiometry of phosphorylation of 0.54 mole of phosphate per mole of EGP‐1. Phosphoamino acid analysis and phosphopeptide mapping of the antigen phosphorylated in vitro by protein kinase C showed that phosphorylation occurred on a serine residue, specifically on serine 303, located in the cytoplasmic domain of EGP‐1. Treatment of ME 180 cells with phorbol ester increased the phosphorylation of EGP‐1. The biological function of EGP‐1 remains to be elucidated. In this report we elucidate an involvement of protein kinase C in phosphorylating EGP‐1, which may signify a role for this antigen in signal transduction across the cell membrane. © 1995 Wiley‐Liss, Inc.
Biochemical and growth‐modulatory effects of the new S‐adenosylmethionine decarboxylase inhibitor CGP 48664 in malignant and immortalized normal human breast epithelial cells in culture
Authors: Manni Andrea , Badger Betty , Wechter Rita , Kunselman Susan , Rossini Anthony , Demers Laurence
Published: February 2007

CGP 48664 [4‐aminoindanon‐1‐(2′‐amidino)hydrazone dihydrochloride monohydrate] is a newly introduced inhibitor of S‐adenosyltnethionine decarboxylase (SAMDC) with increased selectivity of action and reduced toxichy. We analyzed the biochemical and antiproliferative effects of this compound in a panel of hormone‐dependent (3 clones of MCF‐7, T47D) and ‐independent (MDA‐MB‐231, BT‐20) human breast cancer cell lines in culture. For comparison, we also tested its effects in the spontaneously immortalized human breast epithelial cell line MCF‐10A. All cell lines were highly sensitive to the growth‐inhibitory effect of CGP 48664 with an IC50 between 0.1 and 0.5 μM. A dose‐dependent bell‐shaped increase in SAMDC was observed in normal and malignant breast cells resulting from enzyme stabilization by the inhibitor as supported by Western blot analysis. While omithine decarboxylase (ODC) activity consistently increased, the effect of CGP 48664 on spermidine J. spermine N′acetyltransferase (SSAT) was variable in the breast cancer cell lines. In contrast, the inhibitor consistently reduced SSAT activity level in the MCF‐10A cell line and its derivative partially transformed by a mutated ras oncogene. As expected, cellular putrescine levels were markedly increased by CGP 48664 administration, whereas spermidine and spermine contents were reduced. However, the degree of reduction was usually only moderate. Furthermore, exogenous polyamine administration was relatively ineffective in rescuing the antiproliferative effect of CGP 48664 in MCF‐7 cells, while exerting a more complete rescue in the MDA‐MB‐231 cell line. We conclude that CGP 48664 exerts a potent growth‐inhibitory effect on mammary cells in culture. However, its action may not always be entirely mediated through the polyamine pathway. © 1995 Wiley‐Liss, Inc.
Oral contraceptives, menopausal estrogens, and the risk of breast cancer: A case‐control study in greece
Authors: Lipworth Loren , Katsouyanni Klea , Stuver Sherri , Samoli Evangelia , Hankinson Susan E. , Trichopoulos Dimitrios
Published: February 2007

In a hospital‐based case‐control study in Athens, we examined the association between the use of oral contraceptives and menopausal estrogens and the risk of breast cancer. Eight hundred and twenty patients with confirmed breast cancer were compared with 795 orthopedic patient controls and 753 healthy visitor controls, matched to the cases by age and interviewer. The data were modeled through logistic regression, controlling for demographic and reproductive variables. Odds ratio 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 elevated among ever‐users of oral contraceptives, regardless of age at diagnosis of breast cancer, duration of oral contraceptive use or timing of use in relation to first full‐term pregnancy. Among peri‐ and post‐menopausal women who ever used menopausal estrogens, with never‐users as the baseline, a statistically significant elevated odds ratio was found after adjusting for age at menopause. © 1995 Wiley‐Liss, Inc.
An in vivo model to compare human leukocyte infiltration in carcinoma xenografts producing different chemokines
Authors: Melani Cecilia , Pupa Serenella M. , Stoppacoaro Antonella , Ménard Sylvie , Colaghi Maria Ines , Parmiani Giorgio , Colombo Mario P.
Published: February 2007

In this study we tested whether the pattern of cytokines expressed by human carcinomas could account for a different in vivo recruitment of leukocyte subpopulations as a part of the anti‐tumor immune response. Two carcinoma cell lines, SK‐ OV‐3 ovary carcinoma and CALU‐3 lung carcinoma, were analyzed by reverse transcriptase‐polymerase chain reaction (RT‐PCR), immunofluorescence and ELISA for the expression and in vrtro production of cytokines with chemotactic, pro‐ inflammatory and growth‐stimulating activity. Although both cell lines displayed a constitutive expression of granulocyte colony‐stimulating factor (G‐CSF), granulocyte macrophage‐ CSF (GM‐CSF), M‐CSF, interleukin (IL‐) la and IL‐8, only CALU‐3 cell line expressed IL‐10, RANTES (Regulated upon Activation, Normal T Expressed and Secreted) and monocyte‐ activating protein (MCP)‐I. MCP‐I and IL‐8 were detected by immunohistochemistry on sections from tumors xenografted in nude mice. To analyze whether the tumor‐released cytokines modulate leukocytes in tumor infiltration, we studied the distribution of human peripheral blood leukocytes injected in the proximity of SK‐OV‐3 and of CALU‐3 tumor xenografts. While SK‐OV‐3 was unable to recruit human leukocytes and appeared to be barely infiltrated by murine CD45+ cells, CALU‐3 appeared to be rapidly and heavily infiltrated by human leukocytes which induced tumor necrosis within 18–24 hr. © 1995 Wiley‐Liss, Inc.
Analysis of the antibody response to immunization with purified O‐acetyl GD3 gangliosides in patients with malignant melanoma
Authors: Ritter Gerd , Ritter‐Boosfeld Erika , Adluri Rita , Calves Michele , Ren Shunlin , Yu Robert K. , Oettgen Herbert F. , Old Lloyd J. , Livingston Philip O.
Published: February 2007

Gangliosides expressed in malignant melanoma are potential targets for immunotherapy. Immunization of melanoma patients with vaccines containing purified GM2 ganglioside has resulted in induction of GM2 antibodies, and high titers of GM2 antibodies have correlated with increased survival. Melanoma ganglioside 9‐O‐acetyl GD3 is another candidate for ganglioside vaccine construction because of its limited expression in normal human tissues. As purification of 9‐O‐acetyl GD3 from human melanoma (9‐O‐acetylated on the terminal sialic acid) is not practical for broad application, we investigated the antibody response of melanoma patients to O‐acetyl GD3 from several additional sources: hamster melanoma (7‐O‐acetyl GO3), bovine buttermilk (mixture of 7‐O‐acetyl GD3, 9‐O‐acetyl GD3 and 7,9‐di‐O‐acetyl GD3) and chemically modified GD3 from bovine brain (9‐O‐acetylated on the subterminal sialic acid). Only immunization with the buttermilk‐derived O‐acetyl GD3 preparation resulted in consistent production of IgM antibodies. However, the induced antibodies reacted with the immunogen and with 7‐O‐acetyl GO3 derived from hamster melanoma but not with 9‐O‐acetyl GD3 or human melanoma cells expressing 9‐O‐acetyl GD3 on their cell surface. In contrast, all O‐acetyl GD3 derivatives used for immunization were recognized by murine MAbs that reacted with 9‐O‐acetyl GD3, and immunization of mice with buttermilk‐derived O‐acetyl GD3 resulted in the production of antibodies that reacted with human melanoma cells expressing 9‐O‐acetyl GD3. Apparently, the human and murine immune systems preferentially recognize different epitopes on these molecules. © 1995 Wiley‐Liss, Inc.
Recurrent cytogenetic alterations of prostate carcinoma and amplification of c‐myc or epidermal growth factor receptor in subclones of immortalized pnt1 human prostate epithelial cell line
Authors: Degeorges Armelle , Hoffschir Françoise , Cussenot Olivier , Gauville Christiana , Le Duc Alain , Dutrillaux Bernard , Calvo Fabien
Published: February 2007

To develop an experimental prostate cancer model, we immortalized normal human prostate adult epithelial cells with SV40 large‐T antigen. Two sublines were derived in culture, namely PNTIA and PNTIB. They retained the characteristics of prostatic epithelial cells, but did not clone in soft agarose. PNTIA occasionally formed undifferentiated adenocarcinoma tumors in nude mice, but only in the presence of matrigel. PNTIA and PNTIB displayed common cytogenetic alterations: a 10q arm deletion, which is a recurrent alteration in prostate carcinoma, chromosome losses and a translocation involving chromosome 5. An extensive study of oncogenic alterations occurring in these cells showed that PNTIA displayed c‐myc gene amplification, forming an hsr on chromosome 4, as well as c‐myc mRNA overexpression, with a faster doubling time (25 hr); moreover, it seemed less sensitive to EGF than PNTIB. PNTIB had a doubling time identical to that of normal cells (48 hr) but displayed EGF receptor gene amplification accompanied by an increased number of EGF binding sites and sensitivity to EGF. Because both cell lines displayed cytogenetic and oncogenic alterations found in prostate cancer, as well as differing malignant potentials, they represent an interesting model for studying the progression of prostate tumors. © 1995 Wiley‐Liss, Inc.
Comparisons of colon–cancer survival among european countries: The eurocare study
Authors: Sant Milena , Capocaccia Riccardo , Verdecchia Arduino , Gatta Gemma , Micheli Andrea , Mariotto Angela , Hakulinen Timo ,   Franco Berrino The Eurocare Working Group
Published: February 2007

Under the aegis of Eurocare, a European Union project to assemble survival data from population‐based cancer registries and analyze them according to standardized procedures, we have investigated and compared colon‐cancer survival in 10 European countries. We analyzed 68,283 colon‐cancer cases diagnosed between 1978 and 1985 and followed for at least 6 years. After calculating relative survival, putative factors prognostic for survival were investigated by univariate and multiple‐regression analyses. Important intercountry colon‐cancer survival differences exist within Europe, which are not explained by methodological differences, nor by demographic confounders. In patients aged 60 to 69, the mean European 5‐year cumulative relative survival was 40%. Switzerland, Finland and The Netherlands had significantly higher 5‐year relative survival, while one area in the UK and Cracow in Poland had significantly lower survival than this European estimate. Prognosis improved over time: from 1978 to 1985, the risk of death was reduced by about 4% per year in all countries studied. Age at diagnosis is inversely related to prognosis. Differences in health provision and hence in quality of care and stage at presentation seem largely responsible for the differences in colon‐cancer survival found in the EUROCARE countries.
The prognostic value of DNA ploidy and S—phase estimate in primary breast cancer: A prospective study
Authors: Dieterich B. , Albe X. , Vassilakos P. , Wieser S. , Friedrich R. , Krauer F.
Published: February 2007

In this prospective study, the independent prognostic value of DNA ploidy in combination with the major clinico‐pathological characteristics (histological grade, nodal status, tumor size, estrogen and progesterone receptor status, number of tumors, multicentricity, lympho‐vascular infiltration) was evaluated in a series of 399 breast‐cancer patients. The mean follow‐up time was 4.5 years. The DNA content was measured using image cytometry on fresh tumor samples. The overall survival of tetraploid and slowly proliferating diploid cases was significantly different compared with that of aneuploid and rapidly proliferating diploid cases (p = 0.0002). Thus, DNA ploidy combined with S‐phase estimate (DNA histogram type) appeared to be good prognostic factors. In a multivariate survival analysis, DNA histogram type was not an independent prognostic factor unless the histological grade was excluded. This effect of DNA histogram type on survival was also observed among patients with grade‐l or ‐II tumors and patients with small tumors. In conclusion, DNA histogram type was a valuable prognostic factor in univariate analysis, and provided independent complementary information for patients considered at low or intermediate risk by classical pathological findings.
Increased risk of cancer in the descendants of syrian hamsters exposed prenatally to diethylnitrosamine (DEN)
Authors: Mohr U. , Emura M. , Kamino K. , Steinmann J. , Kohler M. , Morawietz G. , Dasenbrock C. , Tomatis L.
Published: February 2007

Transmission of site‐specific tumorigenicity (papillomas in larynx and trachea) of diethylnitrosamine (DEN) to the 2 subsequent generations (F1 and F2) was studied using an out‐bred strain (Han:AURA) of pregnant Syrian golden hamsters (P generation), which were treated i.p. with 10 mg/kg b.w. of DEN on day 12, 13 or 14 of gestation. Laryngotracheal papillomas were induced by DEN in the P and F1 generations only, while these tumours did not occur in the F2 generation. Spontaneously occurring tumours, including uterine adenocarcinomas, lymphomas, and laryngotracheal neuro‐endocrine cell tumours, were observed at higher incidences among the F2 animals derived from the P generation hamsters treated with DEN only on day 13 or 14 of gestation. In the same animals, the ratio of malignant to benign tumours was considerably higher than in controls. In addition, the F2 hamsters derived from the DEN‐treated P generation showed more frequent multiple organ involvement in tumorigenesis than the F2 controls. Several uncommon malignant tumours were detected in the F2 offspring, possibly the result of damage caused to germ cells by the prenatal exposure of F1 Syrian hamsters to DEN.
Anti—vascular approaches to solid tumour therapy: Evaluation of vinblastine and flavone acetic acid
Authors: Hill Sally A. , Sampson Louise E. , Chaplin David J.
Published: February 2007

Several agents have now been identified which exert their anti‐tumour effects in large part via the tumour vasculature; these include TNFα and flavone acetic acid (FAA). More recently, Vincristine and Vinblastine have also been shown to cause a prolonged and selective decrease in tumour perfusion. Vinblastine, unlike FAA, causes no increase in plasma TNFα levels in mice bearing the CaNT tumour, suggesting 2 distinct mechanisms of anti‐vascular activity for these structurally diverse agents. Since FAA and Vinblastine also show quite different normal tissue toxicities, which are separately dose‐limiting, we have examined the strategy of combining these 2 agents. When Vinblastine preceded FAA by 24 hr or less, tumour growth delay was significantly enhanced without a concomitant increase in toxicity. The level of enhancement was not significantly reduced by a 5‐fold decrease in Vinblastine dose, though any reduction in the dose of FAA caused a rapid reduction in treatment effectiveness. Investigation of the functional vasculature of treated tumours suggested that increased anti‐vascular effects may contribute to the enhanced growth inhibition of the combined treatment. Our results demonstrate the potential benefit of combining 2 different classes of antivas‐cular agent, using Vinblastine and FAA (or 5,6‐MeXAA) as prototype drugs.
Improved stage at diagnosis of cervical cancer with increased cancer awareness in a rural Indian population
Authors: Jayant Kasturi , Rao R. S. , Nene B. M. , Dale P. S.
Published: February 2007

As part of its strategy to identify cancer cases in a rural population, the cancer registry of Barshi, India, has developed a methodology which includes education of the population about likely symptoms of cancer, and motivation of symptomatic individuals to undergo medical investigation. Patients with cervical cancer from the registry area who attended Barshi Cancer Hospital (84% of the total) showed a significant improvement in stage at diagnosis between 1988–1989 (38% in stages I and II) and 1990–1992 (51% in stages I and II). No change was observed in those attending the same hospital from a control area (38% vs. 34%). Among a random sample of the population of the registry area, 76% of women were aware of the symptoms of cervical cancer, compared with 25% of the population of control areas. It is suggested that action to raise awareness of symptoms of cancer, and to encourage medical consultation, should form an important initial component of cervical‐cancer control programmes.