bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased levels of 5‐fluorouracil‐induced dna lesions in bloom's syndrome
Authors: Lönn Ulf , Lönn Sigrid , Nylen Urban , Winblad Gerard
Published: February 2007

In Bloom's syndrome (BS) the regulation of uracil‐DNA glycosylase, an enzyme involved in the repair of DNA containing 5‐FU, is altered. 5‐FU induces higher levels of DNA fragmentation in BS cells than in non‐BS cells. The increase in DNA fragmentation is connected to the cytotoxic mechanism where 5‐FU is incorporated into DNA. When 5‐FU induces DNA fragmentation by a mechanism not involving the incorporation of drug into DNA, the levels of DNA fragmentation in BS and non‐BS cells remain similar.
Changes in the expression of idiotype antigen on murine B‐cell lymphoma after hyperthermia alone and in combination with interferon and tumour necrosis factor
Authors: Davies Catharina De L. , Basham Teresa Y. , Anderson Robin L. , Hahn George M.
Published: February 2007

The effect of interferon (IFN) and tumour necrosis factor (TNF), either alone or combined with hyperthermia, on cell proliferation and expression of idiotype antigen on a murine B‐cell lymphoma has been studied. Incubation with same doses of IFN‐α and IFN‐γ reduced cell proliferation to the same extent. Hyperthermia potentiated the antiproliferative activity of IFN‐α and IFN‐γ. Pretreatment with IFN‐γ induced a synergistic response with heat, while IFN‐α and heat had an additive effect. Tumour necrosis factor (TNF) alone did not affect cell proliferation, nor did TNF modify the heat‐induced delay in cell growth.

The quantitative expression of surface idiotype antigen was studied by flow cytometry using an anti‐idiotype monoclonal antibody (MAb). Heat reduced the expression of idiotype antigen approximately 50%. The duration of the reduction depended on the heat‐dose. Recovery of antigen expression correlated with recovery of cell growth, and 2–5 days after the treatment antigen expression returned to the normal level for untreated cells.

IFN‐γ and TNF increased antigen expression (30–50%) which lasted for 4–6 days after treatment. When cells were incubated with IFN‐γ or TNF for 2 days prior to hyperthermia, the increase in antigen expression was observed immediately after heating, but by the following day, antigen expression was similar to that after heat treatment alone. Expression of idiotype antigen recovered within 2–5 days to the same values as after heat treatment alone. IFN‐α alone or combined with hyperthermia did not have any significant effect on antigen expression.
Mapping of linear epitopes of human papillomavirus type 16: The L1 and L2 open reading frames
Authors: Dillner Joakim , Dillner Lena , Utter Göran , Eklund Carina , Rotola Antonella , Costa Silvano , Diluca Dario
Published: February 2007

Certain types of human papillomavirus (HPV), notably HPV type 16, are associated with flat or inverted proliferative lesions of the cervix uteri that can progress to malignancy. As a first step towards the serological study of the epidemiology of HPV, we have synthesized the entire amino acid sequences of the 2 major viral capsid proteins of HPV type 16, L1 and L2, as a set of 66 synthetic 20‐residue peptides with an overlap of 5 amino acids. The peptides were tested for reactivity with IgA, IgG and IgM antibodies in the sera of 30 patients with HPV‐16‐carrying cervical neoplasms. Both IgG and IgM antibody responses were detected, but most of the reactivity found was of the IgA class. The most immunoreactive peptides were futher analyzed for reactivity with sera from 22 patients with parotid gland tumors and with sera from 38 healthy individuals. The L2‐encoded protein contained only one major linear epitope, which was not specific for HPV‐16‐carrying neoplasms. In contrast, the L1‐encoded protein contained several epitopes that were regularly immunoreactive with antibodies present in the sera of patients with HPV‐16‐carrying cervical neoplasms, but only rarely so in the sera of patients with other tumors or of healthy individuals.
Suramin inhibits proliferation of rat glioma cells and alters N‐cam cell surface expression
Authors: Fantini Jacques , Guo Xiao‐Jun , Marvaldi Jacques , Rougon Geneviève
Published: February 2007

Suramin, a drug used in the treatment of trypanosomiasis and onchocerciasis inhibits growth‐factor‐induced mitogenesis. We have investigated the effect of suramin on the growth rate and the morphology of C6 glioma cells cultured in the presence of serum or in a serum‐free defined medium. Exponentially growing cells were seeded in multi‐dish plates (5 × 104 cells/2 cm2 well) in DMEM supplemented with 5% fetal calf serum and were continuously exposed to 1 μg/ml to 1,000 μg/ml suramin. Growth rate (determined 9 days after seeding) was reduced by 5%, 33%, 56% and 97%, respectively for suramin concentrations of 1, 10, 100 and 1000 μg/ml. Similar results were obtained in serum‐free defined medium (DMEM/F12, 1:1, v:v, EGF 5 ng/ml, transferrin 5 μg/ml, selenium 10 ng/ml). Moreover, the concentration of suramin in the culture medium remained constant, demonstrating that the drug was not actively metabolized by the cells. Suramin also induced marked changes in cell morphology: the usual bipolar shape of C6 cells evolved toward a more differentiated appearance, with numerous cellular processes allowing a wide number of cell‐cell contacts. In parallel, we monitored expression of an adhesion molecule (N‐CAM) at both the mRNA and protein levels. Indirect immunofluorescence technique showed an important increase in cell surface N‐CAM expression, starting from a dose of 10 μg/ml suramin, whereas total cellular content of N‐CAM protein as well as its mRNA levels were unaffected. We also observed that the levels of expression of actin and N‐CAM mRNAs decreased by a factor of two in cells maintained in defined medium. However, the relative ratio of N‐CAM mRNA over actin mRNA was virtually unchanged following suramin treatment. Taken together, our results suggest that suramin (i) exerts a blocking effect of autocrine growth factors, (ii) interferes with the turn‐over mechanisms of N‐CAM expressed at the cell surface, either by impairing its endocytosis and/or the process of release of the N‐CAM 120 isoform.
Deregulated c‐myc expression in epstein‐barr‐virus‐immortalized b‐cells induces altered growth properties and surface phenotype but not tumorigenicity
Authors: Hotchin Neil A. , Allday Martin J. , Crawford Dorothy H.
Published: February 2007

Endemic Burkitt's lymphoma (eBL) is characterized by the presence of Epstein‐Barr virus (EBV) and a chromosomal translocation which results in deregulation and constitutive expression of the c‐myc proto‐oncogene. In order to examine the role played by activation of c‐myc in determining the eBL phenotype, we have introduced into EBV‐immortalized lymphoblastoid cells (LCL) plasmids which permit constitutive expression of c‐myc. The resulting cells show a reduced serum dependence, reduced homotypic cell aggregation, and changes in surface characteristics. In particular, levels of the cell adhesion molecule, LFA‐1, are greatly reduced. However, the cells continue to express all the EBV latent antigens associated with the LCL phenotype and they remain nontumorigenic. These results suggest that, whilst constitutive expression of c‐myc may contribute to the malignant phenotype, it is insufficient to induce tumorigenicity.
Neoplastic transformation of a human kidney epithelial cell line transfected with V‐HA‐ras oncogene
Authors: Haugen Aage , Ryberg David , Hansteen Inger‐Lise , Amstad Paul
Published: February 2007

We have recently shown that normal human kidney epithelial (NHKE) cells were immortalized by treatment with Ni(II) alone. In the present study the immortalized human kidney cell line (IHKE) was transfected with a plasmid construct containing the v‐Ha‐ras oncogene (pZipras). After transfection, the cell lines formed tumors in athymic nude mice, whereas the ZipNeoSV(X)‐transfected IHKE control cultures formed no tumors. Tumor cell lines (THKE) were established from the tumors in nude mice. These cells appear to be of human epithelial origin and express high levels of Ha‐ras transcript. Karyotypic analysis was performed. The cell lines were tritetra‐ or pentaploid. A consistent finding in the IHKE, IHKZE and THKE cells was increased numbers of chromosomes 17 and 7p+. Some marker chromosomes were identical in the IHKE and THKE cell lines, underlining their common origin and their possible importance in the carcinogenic process. This shows that the combined action of a chemical carcinogen [i.e., Ni(II)] and v‐Ha‐ras oncogene resulted in fully transformed human kidney epithelial cells, consistent with a stepwise progression of human epithelial cell transformation.
Tumor progression by dna flow cytometry in human colorectal cancer
Authors: Giaretti Walter , Santi Leonardo
Published: February 2007

We wished to better understand the role of aneuploidy during the progression of human colorectal cancer. Fresh or frozen multiple samples from 221 human colorectal adenomas, 93 carcinomas, and corresponding control mucosa were investigated using high‐resolution DNA flow cytometry. A total number of 164 DNA abnormal clones were observed and characterized by a quantitative index of DNA aneuploidy (DI). In precancerous lesions the vast majority of DNA abnormal clones (almost 3/4 in adenomas with mild to moderate dysplasia) was hypo‐ and hyper‐diploid with DI values from 0.8 to 1.2 (near‐diploidy). In moderately to poorly differentiated carcinomas the vast majority of abnormal clones was near‐triploid and hypotetraploid with DI values from 1.4 to 1.8 (near hypertriploidy) and only 12% were near‐diploid. Adenomas with foci of carcinomas, a group of special interest since they represent a link in colorectal tumor progression, had median triploid DNA content. In addition to an increase in DI values, the carcinomas had a clear increase in the proportion of cells actively synthesizing DNA (S‐phase fraction). These results are interpreted as evidence for a ploidyevolution model according to which near‐diploid clones in adenomas at early stages of dysplasia would derive from abnormal mitotic cells that divide their DNA unequally between two daughter cells. Tetraploidization of these near‐diploid cells and successive DNA loss would then lead in later stages of tumor progression to near‐hypertriploid clones characterized by a balance of chromosomes bearing growth‐promoting and growth‐suppressing genes confering a selective proliferative advantage.
Heterogeneity of immunoglobulin gene rearrangements in B‐cell lymphomas
Authors: Kneba M. , Bergholz M. , Bolz I. , Hulpke M. , Bätge R. , Schauer A. , Krieger G.
Published: February 2007

We have examined 69 B‐cell non‐Hodgkin's lymphomas (NHL) for rearrangements of the immunoglobulin (Ig) or T‐cell antigen receptor (TCR) genes. The lymphomas were assigned to the categories of the Kiel classification and their B‐cell nature was confirmed by immunostaining. Only 2 cases (with CLL) displayed clonal Tβ‐chain TCR gene rearrangements together with rearranged heavy‐ and light‐chain Ig genes. The remaining 67 lymphomas had a germline β‐chain TCR‐gene configuration. Three different patterns of Ig gene rearrangements were identified; (A) presence of both heavy‐ and light‐chain rearrangements (H+L+); (B) rearrangement of heavy‐chain gene only (H+L−); (C) heavy‐ and light‐chain genes in germline configuration (H−L−). All the 45 low‐grade NHLs and the 4 immunoblastic lymphomas exhibited pattern A and all had their kappa gene rearranged or deleted. Of 24 low‐grade lymphomas tested, 13 (54%) had an additional rearrangement of the lambda light‐chain gene. In contrast, the 19 high‐grade centroblastic (cb) B‐NHLs had distinct patterns of Ig‐gene rearrangement: 12 with pattern A, 4 with B and 2 with C. In this group only 2 of 17 (12%) cases analyzed had evidence of lambda light‐chain rearrangement whereas 12 of 18 (67%) had a kappa gene rearrangement or deletion. In one case expressing sIgM/lambda and with heavy chain Igrearrangement, no DNA was available for Ig light‐chain analysis.
Prevalence of cervical dysplasia and HPV infection according to sexual behavior
Authors: Azocar Jose , Abad S. M. J. , Acosta Humberto , Hernandez Rosa , Gallegos M. , Pifano E. , Blanch Ricardo , Kramar Andrew
Published: February 2007

A total of 165 uterine cervix smears from Venezuelan women were examined by cytological techniques to identify malignant and pre‐malignant cervical changes, as well as to identify Human Papilloma Virus (HPV) DNA types 6, 11, 16, 18, 31, 33 and 35. Of these smears, 119 were from nonmonogamous women who participated in a cervical carcinoma screening program. In this group, HPV‐DNA was detected by hybridization in 42 samples (35%) and cervical intraepithelial neoplasia (CIN) in 13 (11%). Forty‐six monogamous aboriginal women were similarly studied and no evidence of abnormal cytology or HPV‐DNA of the types studied here was found in any of them. In the non‐monogamous group, age at first sexual intercourse and index of parity were not associated with cervical HPV infection and/or CIN. The rates of HPV infection, however, were significantly different between the two populations, confirming that sexual behavior involving multiple partners is associated with HPV infection.
The capsule surrounding primary liver tumors: Wherefrom its prognostic significance?
Authors: Grigioni W. F. , D'Errico A. , Biagini G. , Mazziotti A. , Bolondi L. , Liotta L. A. , Mancini A. M. , Garbisa S.
Published: February 2007

Primary liver neoplasms in cirrhotic and non‐cirrhotic patients were studied by electron microscopy and immunohistochemical methods for extracellular matrix (ECM) antigens. A capsule of variable thickness was present in many expanding hepatocellular carcinomas, while it was absent in those of small size, and either fragmented or absent in the infiltrating ones. In the capsules of early onset, fibronectin was the most frequent stromal glycoprotein. In the completely formed capsular structures, fibronectin, type‐V collagen and laminin were the most common macromolecules seen. No differences were evident in the pattern of ECM macromolecules in the capsules surrounding hepatocellular carcinomas compared with those found in benign lesions, such as hepatocellular adenomas or focal nodular hyperplasia. The possibility is discussed that the capsule could be a result of the following sequence: tissue with expansive (not infiltrative) growth–mechanical compression–ischemic necrosis of surrounding tissues–repair process with ECM deposition.
The distribution of non‐burkitt, non‐Hodgkin's lymphomas in Uganda in relation to malarial endemicity
Authors: Schmauz R. , Mugerwa J. W. , Wright D. H.
Published: February 2007

Biopsies of malignant lymphomas collected from all districts of Uganda, filed in the Kampala Cancer Registry for the 8‐year period 1966–1973, were reviewed. This review confirmed a relatively low frequency of follicle‐centre‐cell lymphomas with a follicular growth pattern and the geographical co‐distribution between malaria and Burkitt's lymphoma (BL). It also showed a similar, though less marked, association between non‐Burkitt, non‐Hodgkin's lymphoma (NBNHL) and malarial endemicity, and a correlation in the regional incidence between BL and NBNHL in both comparisons, these associations were strong for high‐grade lymphomas and weak for low‐grade neoplasms. BL and other NHL may therefore share, to a varying degree, some common pathogenesis. The excess in frequency of NBNHL of high‐grade malignancy in malarial endemic areas appears to be in contrast to Western countries where most non‐Hodgkin's lymphomas are of low‐grade malignancy.
DNA adducts in different tissues of smokers and non‐smokers
Authors: Cuzick Jack , Routledge Michael N. , Jenkins David , Garner R. Colin
Published: February 2007

Purified DNA from human lung, liver, bladder, pancreas, breast and cervix has been analysed for DNA adducts using the nuclease P1 modification of the 32P post‐labelling technique. Tissues were obtained at autopsy from 13 men and 6 women. Relatives were asked to provide information on smoking history for deceased subjects. All tissues examined except the breast had detectable adducts. In lung, bladder and pancreatic tissue a characteristic pattern of adducts was seen which has previously been reported as typical of cigarette‐smoke‐induced damage. Smokers and former smokers tended to have higher adduct levels than non‐smokers in the tissues examined but this was only significant for the lung. There appeared to be considerable variation in adduct levels among smokers which could not be accounted for by duration or daily consumption level. Certain smokers had high adduct levels in all tissues examined, whilst in others high levels were only seen in some tissues. All cervical samples examined had detectable adducts. These results confirm the finding that cigarette smoking is associated with DNA damage in the lung and suggest that similar damage may be related to tobacco‐induced neoplasms of other tissues.
Morphologic analysis of human lymphokine‐activated killer (LAK) cells
Authors: Groscurth P. , Diener S. , Stahel R. , Jost L. , Kägi D. , Hengartner H.
Published: February 2007

Lymphokine‐activated killer (LAK) cells obtained from normal donors at various days of in vitro cultivation have been studied by several methods including scanning (SEM) and transmission (TEM) electron microscopy, immuno‐electron microscopy, in situ hybridization and flow cytometric DNA measurements. In addition, the cytotoxic activity of LAK cells against several tumor cells was examined by 51Cr‐release assay and by SEM and TEM. The LAK cells displayed a uniform ultrastructural appearance concerning surface structure and morphology of organelles. They contained typical lysosomal granules which by immuno‐electron microscopy showed a specific localization of perforin 1 (P1). The presence of P1 and granzymeA mRNA in the cytoplasm was confirmed by in situ hybridization using specific antisense probes. Frequency and increase of specific mRNA‐containing cells was similar for both genes. Single LAK cells were further characterized by peculiar nuclear inclusion bodies (IB) which were presumably formed by trapped profiles of endoplasmic reticulum. Flow cytometric analysis revealed normal DNA content of LAK cells even after prolonged cultivation indicating that the IB were not associated with aneuploidy of the effector cells. The LAK cells were highly effective in lysing K562 and DAUDI cells as shown by 51Cr‐release assay. They caused characteristic morphologic alterations of target cells similar to those found in cytotoxic T‐lymphocyte (CTL) and NK‐cell‐mediated cytolysis. SEM and TEM studies on specimens prepared by routine procedures or by cryopreparation showed that the tumor cell membrane was the initial target for the LAK cell attack whereas other cell compartments were damaged only in advanced stages of cytolysis. Summarizing our study demonstrates that LAK cells have a characteristic ultrastructure which in some aspects differs from that of CTL and NK cells, and that LAK cells appear to destroy tumor cells by mechanisms similar to those of other cytotoxic effector cells.
Characterization of an embryonal rhabdomyosarcoma cell line showing amplification and over‐expression of the N‐myc oncogene
Authors: Hayashi Yasuhide , Sugimoto Tohru , Horii Yoshihiro , Hosoi Hajime , Inazawa Johji , Kemshead John T. , Inaba Toshiya , Hanada Ryoji , Yamamoto Keiko , Gown Allen M. , Sawada Tadashi
Published: February 2007

A parent rhabdomyosarcoma cell line designated SCMC‐RM2 was established from bone‐marrow tumor cells taken from an 11‐year‐old girl with an embryonal rhabdomyosarcoma. Subsequently a cloned SCMC‐RM2‐1 cell line was isolated from a parent line. These cell lines grew as adherent monolayers in liquid culture with a doubling time of 50 and 52 hr, respectively. In addition, colonies were established in soft agar, which grew in a dose‐dependent fashion with a cloning efficiency of 0.7 and 0.8%, respectively. Chromosomal analysis showed these cell lines had neither double minutes nor homogeneously staining regions. Chromosome number ranged from 61 to 93, translocation; t(9;13)(p22;q14) was identified, and no alteration of chromosome 2 was observed. Surface membrane antigen profile of parent and cloned lines by using a panel of 24 monoclonal antibodies (MAbs) excluded the possibility of these being neuroblastoma cell lines. In addition, MAbs to the cytoplasmic protein desmin, myoglobin, muscle actin (α and γ) and α‐sarcomeric actin reacted with these cell lines, SCMC‐RM2 and SCMC‐RM2‐1 being thus identified as rhabdomyosarcoma. Southern blot analyses revealed 8‐ and 7‐fold amplification of the N‐myc gene in SCMC‐RM2 and SCMC‐RM2‐1 as compared with the promyelocytic cell line HL60. Over‐expression of the N‐myc mRNA was noted over control cell lines.
Differentiation‐inducing and growth‐inhibitory activities of erythroid differentiation factor (EDF/activin A) toward mouse erythroleukemic cells in vivo
Authors: Shiozaki Makoto , Shioya Shigeru , Izawa Masaaki , Eto Yuzuru , Shibai Hiroshiro
Published: February 2007

The in vivo differentiation‐inducing activity of a purified human erythroid differentiation factor (EDF) toward mouse erythroleukemic cells (MEL cells) was examined. BDF1 mice with diffusion chambers implanted in the peritoneal cavity were treated with continuous i.p. administration of EDF. MEL cells within a diffusion chamber differentiated into hemoglobin‐positive cells when treated with EDF, the percentage of the positive cells being 32.3 ± 28.3 on day 5 as compared to 0.2 ± 0.3 in the controls. The anti‐tumor activity of EDF was also examined in a nude mouse MEL solid tumor model. Daily intra‐tumor treatment with EDF for 10 days resulted in 73% suppression of tumor growth on day 25. A histological study revealed that EDF‐treated solid tumor cells became hemoglobin‐positive, indicating the anti‐tumor activity of EDF through induction of differentiation in vivo. EDF could induce in vitro the differentiation of human erythroleukemic cell lines K562 and HEL, as well as the murine cell line. These results indicate the possibility of differentiation therapy for erythroleukemia using EDF.
Enhanced expression of the proto‐oncogenes fos and raf in the rhabdomyosarcoma cell line BA‐HAN‐1C after differentiation induction with retinoic acid and N‐methylformamide
Authors: Gabbert N. E. , Gerharz C.‐ D. , Ramp U. , Hoffmann J. , Oster O. , Oesch F. , Doehmer J.
Published: February 2007

BA‐HAN‐1C is a clonal rat rhabdomyosarcoma cell line consisting of proliferating mononuclear tumor cells, some of which spontaneously fuse to form terminally differentiated post‐mitotic myotubes. Exposure of BA‐HAN‐1C cells to retinoic acid (RA) or N‐methylformamide (NMF) resulted in a significant inhibition of proliferation (p < 0.001) and in cellular differentiation, as evidenced by a significant increase in the creatine kinase (CK) activity (p < 0.05) and the number of terminally differentiated post‐mitotic myotubes (p < 0.001). Furthermore, between 5% (NMF) and 30% (RA) of the mononuclear tumor cells exhibited ultrastructural features of rhabdomyogenic differentiation, not observed in their mononuclear counterparts under standard growth conditions. Although BA‐HAN‐1C cells responded to both inducers of differentiation, differences in time course and magnitude of both increase of differentiation and growth inhibition were observed. These effects of RA and NMF were preceded by a marked enhancement of c‐raf expression which became evident 6 and 12 hr after exposure to RA and NMF, respectively, and which persisted throughout the observation period of 5 days. Furthermore, a transient expression of c‐fos could be observed 15 and 30 min after exposure to RA. Our results suggest that c‐raf expression might be implicated in the differentiation process of BA‐HAN‐1C tumor cells.
In vitro and in vivo anti‐tumor activity of L‐glutamic acid γ‐monohydroxamate against L1210 leukemia and B16 melanoma
Authors: Vila Jorge , Thomasset Nicole , Navarro Claudine , Doré Jean‐François
Published: February 2007

A glutamine analogue, L‐glutamic acid γ‐monohydroxamate (GAH) demonstrated complete cytotoxicity against L1210 cells in culture and marked anti‐tumoral activity in vivo against L1210 leukemia and B16 melanoma. In vitro, GAH caused concentration‐dependent inhibition of L1210 cell growth, with complete cell death being reached at 72 hr and at a 500 μM concentration. A minimal incubation time of 38 hr with 500 μM GAH was necessary to obtain complete cell death at 72 hr. During incubation, GAH is metabolized to hydroxylamine. Hydroxylamine acts as the active form of GAH, since the concentration‐dependent inhibition of cell growth caused by hydroxylamine is the same as that observed with GAH. The cytotoxic effects of GAH and hydroxylamine on L1210 cells were not reversed or prevented by L‐glutamine or L‐glutamic acid and purine nucleosides but were prevented or reversed by pyruvate, 2‐oxaloacetate and 2‐oxoglutarate.

In vivo, GAH considerably increased survival of mice bearing L1210 leukemia or a solid tumor, the B16 melanoma. Anti‐tumor activity of GAH against L1210 leukemia and B16 melanoma was schedule‐dependent. The administration of GAH 3 times daily was more effective than a twice daily treatment and the maximum ILS was observed using split‐dose schedules on days 1 through 3 and 7 through 9 without noticeable toxicity. Under these conditions hydroxylamine is highly toxic, suggesting that in vivo GAH might act as an hydroxylamine releaser in the tumor cells and is not significantly metabolized in the body.
Establishment and characterization of a continuous in vitro line from a rat choriocarcinoma
Authors: Verstuyf A. , Sobis H. , Goebels J. , Fonteyn E. , Cassiman J. J. , Vandeputte M.
Published: February 2007

A continuous in vitro cell line of rat choriocarcinoma has been established. It is composed of pure trophoblast cells which multiply and differentiate. The morphology of the cells is very similar to normal rat cytotrophoblasts and giant cells. The cultured cells contain cytokeratin, alkaline phosphatase and express the receptors for Bandeira simplificifolia Agglutinin‐1 (BSA‐1). They are hormonally active as demonstrated by the presence of lactogen and progesterone in the supernatant of the culture. The injected cells develop into choriocarcinoma in syngeneic as well as allogeneic rats. The morphological, biological and immunohistochemical features of these tumors are identical to those described in the transplantable neoplasm from which the in vitro line was established. The presence of γ chromosome in cultured cells proves the paternal origin of the primary tumor developed from extra‐embryonic membranes in fetectomized rat and makes this neoplasm similar to human post‐gestation choriocarcinoma.
Western blotting analysis for malignant lymphoma and stomach cancer antigens from carcinogen‐transformed bloom syndrome cells
Authors: Shiraishi Yukimasa
Published: February 2007

Membrane antigen (glyco‐) proteins associated with malignant lymphoma (ML) and stomach (ST) cancer were collected from a carcinogen‐transformed Bloom syndrome (BS) B‐lymphoblastoid cell line by means of the panning protocol, in which antigen cell clones which specifically react with antibodies in ML and ST cancer sera were separated on polystyrene dishes coated with IgG. Gel electrophoresis and Western blotting (WB) analyses demonstrated that antigens which characterize ML and ST cancer gave a single band (ML, 97 kDa; ST, 118 kDa) and did not cross‐react with sera of other cancer patients or with normal sera. The present WB procdure may be useful for diagnostic purposes. The amino acid sequence of the ML antigen band (97 kDa) had 76.9% (in 13 sequences) homology to that of the variable region of the immunoglobulin (Ig) heavy chain (IgVH) at the N‐terminal end, while the sequence of the ST (118 kDa) cancer antigen had little homology to IgVH.
Neoadjuvant immunotherapy with interferon of the spontaneously metastasizing murine B16F10L melanoma
Authors: Markovic Svetomir N. , Murasko Donna M.
Published: February 2007

We have previously demonstrated that administration of interferon a/b (IFN) for 4–5 days after challenge with a transplantable Moloney sarcoma virus‐induced tumor completely inhibited tumor development. In the present study, we examined the therapeutic effects of IFN on mortality induced by metastatic dissemination of the B16F10L murine melanoma. IFN was administered at various times in relation to the surgical removal of primary tumor: days ‐5 to ‐1 prior to tumor excision (neo‐adjuvant protocol), or for 5 days after tumor excision, beginning on days 1, 6 or 11 after excision of the primary tumor (adjuvant protocols). The neo‐adjuvant protocol was superior to all other protocols, significantly increasing percentage survival (56% vs. 0%) and median survival time (>84 days vs. 33 days) compared to untreated controls, as well as to all adjuvant protocols. In contrast, IFN treatment on days 1 to 5 after excision of the primary tumor decreased median survival time of cases compared to untreated controls (20 days vs. 33 days). Both IFN‐induced inhibition and enhancement of metastatic dissemination were dose‐dependent, with higher amounts of IFN producing greater inhibition or enhancement. The superior therapeutic efficacy of the neo‐adjuvant IFN treatment was associated with increased spleen and lung‐derived natural killer cell cytolytic activity (on days ‐4, 0 and 2) followed by a later (day 13) increase in lung‐associated cytolytic T‐cell responses.
Fatty acid composition of phospholipids in erythrocyte membranes and risk of breast cancer
Authors: Zaridze D. G. , Chevchenko V. E. , Levtshuk A. A. , Lifanova Y. E. , Maximovitch D. M.
Published: February 2007

Five fatty acids (FA) 2 saturated, palmitic (C16:0), stearic (C18:0) and 3 unsaturated, oleic (C18:1), linoleic (C18:2), arachidonic (C20:4) acids have been studied in the red cell membranes of breast cancer patients and controls. Statistically significant decrease in the risk of breast cancer has been found to be associated with increase in the levels of linoleic acid in pre‐menopausal women and of arachidonic acid in the post‐menopausal group.
Bronchial micronuclei as a marker of an early stage of carcinogenesis in the human tracheobronchial epithelium
Authors: Lippman Scott M. , Peters Edward J. , Wargovich Michael J. , Stadnyk Alexander N. , Dixon Dennis O. , Dekmezian Roupen H. , Loewy John W. , Morice Rodolfo C. , Cunningham Joan E. , Hong Waun K.
Published: February 2007

In the bronchial epithelium, smoking initiates a multistep process that first appears histologically as premalignant squamous metaplasia/dysplasia, a biological predecessor of squamous‐cell lung cancer. Reflecting chromosomal damage from a carcinogenic insult, micronuclei may reveal earlier events in the carcinogenic sequence. We prospectively evaluated and correlated micronucleus count, histology (index of metaplasia) and smoking exposure in 35 consecutive subjects (9 active smokers, 10 previous smokers and 16 never‐smokers) undergoing diagnostic bronchoscopy. Samples for micronuclei and histological evaluation were taken from the main carinal mucosa in each subject for site‐specific comparisons. The median and mean micronucleus counts per 1,000 cells were significantly higher in active smokers than in non‐smokers (subjects who had never smoked and previous smokers): median counts were 3.7 vs. 1.4, p = 0.03; mean counts were 4.7 vs. 1.9, p = 0.01. There was no significant difference, however, in micronucleus counts between subjects who had never smoked and previous smokers. Bronchial metaplasia and smoking history were not associated. Our findings suggest that micronuclei are a readily quantitated, early intermediate‐endpoint marker for detecting tobacco‐initiated tracheobronchial carcinogenesis.
Dietary fiber, beta‐carotene and breast cancer: Results from a case‐control study
Authors: Veer Pieter Van 'T , Kolb Corine M. , Verhoef Petra , Kok Frans J. , Schouten Evert G. , Hermus Rudolph J. J. , Sturmans Ferd
Published: February 2007

To study the association between dietary fiber, betacarotene and breast cancer, the average daily intake of these dietary components was compared among 133 incident breast cancer cases and 238 population controls. Average daily intake of cereal products, fruit and vegetables was also studied. A statistically significant lower energy‐adjusted intake of dietary fiber was observed in cases than in controls (mean ± SD: 25.4 ± 6.7 g vs. 27.7 ± 7.4 g, 95% confidence interval (CI) of the age‐adjusted difference = −3.8, −0.8). Intake of betacarotene was similar for cases and controls. The multivariate adjusted odds ratio (OR) of breast cancer among women in the highest quartile of intake of cereal products, as compared to those in the lowest quartile, was 0.42 (95% CI = 0.19 – 0.92) and the trend was statistically significant (p = 0.03). The corresponding OR for intake of dietary fiber was 0.55 (95% CI = 0.26 – 1.17) but the trend was not significant. The OR for the highest quartile of intake of beta‐carotene, fruit, vegetables, and all vegetable products combined was less than unity, but there was no significant inverse trend. These results suggest that a high intake of cereal products, especially those rich in fiber, may be inversely related to incidence of breast cancer.
Alcohol, beer and cancer of the pancreas
Authors: Bouchardy Christine , Clavel Francoise , La Vecchia Carlo , Raymond Luc , Boyle Peter
Published: February 2007

The relationship between total alcohol consumption and intake of beer, wine and spirits and the risk of cancer of the pancreas was re‐assessed in a pooled analysis of 3 case‐control studies of pancreatic cancer from Italy, France and Switzerland, providing a total data‐set of 494 cases and 1,704 controls. Logistic regression was used to obtain relative risks adjusted for study, age, sex, smoking and socio‐economic status. Relative to non‐drinkers, the risk estimates for subsequent levels of alcohol consumption were close to unity, and there was no evidence of a trend in risk with dose; the point estimate for more than 8 drinks per day was 0.8(95% Confidence Interval, Cl = 0.5–1.3). Likewise, no consistent association was observed for consumption of wine, beer or spirits: the relative risks for the highest consumption levels were 1.0 for wine, 0.9 for beer and 0.9 for spirits. No significant interaction was observed with study centre, sex or smoking habits. Given the large size of the data‐set, of the consistency and the replication of findings across the 3 different studies, and of the elevated alcohol consumption of these populations, the present analysis gives reassuring evidence on the alcohol and pancreatic cancer issue in relation to total consumption of alcohol, of beer and of other alcoholic beverages.
pp60c‐src protein kinase activity in human gastric carcinomas
Authors: Takekura Naoki , Yasui Wataru , Yoshida Kazuhiro , Tsujino Tetsuhiro , Nakayama Hirofumi , Kameda Takashi , Yokozaki Hiroshi , Ishimura Yasuhiko , Ito Hisao , Tahara Eiichi
Published: February 2007

We examined pp60c‐src protein kinase activity in human gastric carcinoma cell lines and gastric carcinoma tissues as well as normal mucosa. pp60c‐src kinase activity was detected in all 5 carcinoma cell lines at various levels. Of 16 gastric carcinoma tissues, 8 showed higher pp60c‐src kinase activity in tumor tissues than in corresponding normal mucosa. However, the levels of expression ofpp60c‐src detected by Western blotting were not always consistent with the activities of pp60c‐src protein kinase. These findings suggest that the increase in pp60c‐src protein kinase activity might be brought about by post‐translational changes.
The consumption of tobacco, alcohol and the risk of adenocarcinoma in Barrett's oesophagus
Authors: Levi Fabio , Olloy Jean‐Baptiste , La Vecchia Carlo , Boyle Peter , Monnier Philippe , Savary Marcel
Published: February 2007

The relationship between tobacco, alcohol and the risk of oesophageal adenocarcinoma in Barrett's oesophagus was evaluated in an endoscopy‐clinic‐based case‐control study of 30 histologically confirmed cases of adenocarcinoma and 140 controls with Barrett's oesophagus but no evidence of malignant lesions. Among the cases, 18 (60%) were non‐smokers and 14 (47%) non‐drinkers, the corresponding proportions in the controls being 52% and 44%. Thus, there was no apparent relation between tobacco, alcohol and the risk of adenocarcinoma of the oesophagus, the age‐ and sex‐adjusted point estimates being 1.0 for moderate and 0.9 for heavy smokers, 0.7 and 1.5 respectively for moderate and heavy drinkers. Upper 95% confidence limits were 1.6 for ever‐use of tobacco and 1.9 for ever alcohol drinking. The findings of this study, although based on a limited number of cases, indicate that alcohol and tobacco are unlikely to play a major role in the aetiology of adenocarcinoma in Barrett's oesophagus.
A case‐control study of gastric cancer and diet in Italy: II. Association with nutrients
Authors: Buiatti Eva , Palli Domenico , Decarli Adriano , Amadori Dino , Avellini Claudio , Bianchi Simonetta , Bonaguri Chiara , Cipriani Francesco , Cocco Pierluigi , Giacosa Attilio , Marubini Ettore , Minacci Chiara , Puntoni Riccardo , Russo Antonio , Vindigni Carla , Fraumeni Joseph F. , Blot William J.
Published: February 2007

A case‐control study involving interviews with 1,016 gastric cancer (GC) patients and 1,159 population‐based controls in high‐ and low‐risk areas was conducted to evaluate dietary factors and their contribution to the marked geographic variation in mortality from this cancer within Italy. Risks of GC were found to vary significantly with estimated nutrient in‐take. Risk rose with increasing consumption of nitrites and protein, and decreased in proportion to intake of ascorbic acid, beta‐carotene, alpha‐tocopherol, and vegetable fat. The associations with nitrite and beta‐carotene tended to fade, however, in multivariate analyses adjusting for intake of other nutrients. Ascorbic acid showed the strongest geographic gradient, with highest consumption in low‐risk areas. The findings suggest that the protective effects we previously reported for consumption of fresh fruit, fresh vegetables and olive oil may be linked to the vitamins C and E contained in these foods. The findings are consistent with the hypothesis that N‐nitroso compounds are involved in GC risks, since elevated risks were apparent for agents (nitrites, protein) that promote nitrosation, while decreased risks were found for nutrients (ascorbic acid and alpha‐tocopherol) which inhibit the process.
Rat monoclonal antibodies produced against rat colorectal adenocarcinomas define tumor‐ and colon‐associated, auto‐immunogenic antigens
Authors: Brodin N. Thomas , Jansson Bo , Sjögren Hans Olov
Published: February 2007

Monoclonal antibodies (MAbs) were derived from rats immunized against allo‐ and syngeneic rat colon carcinomas. Screening was performed by immunohistochemistry modified for MAbs of the same species as the tissue used for frozen sections. From 2 fusions, several MAbs were found that bound to syngeneic tumor tissue but showed little or no staining of normal adult tissues. Another group of MAbs demonstrated an intracellular staining of tumor cells and staining of mucus and goblet cells in the distal 2/3 of the normal colon. A third group of MAbs showed staining of both tumor and normal colon tissue. Staining patterns were reproduced with 6 MAbs selected from the first 2 groups after purification and biotinylation. One MAb, 10B 12, recognized an antigen expressed in 10/11 colon carcinomas, the lowest parts of colonic crypts, intracellularly in a subpopulation of pancreatic acinar cells and mucus in antrum. It was used for affinity purification of a high‐molecular‐weight antigen, to which 3 of the other rat MAbs also bound. This antigen was also bound by antibodies to blood group A and isogloboseries carbohydrate determinants. Competition with the labelled 10B12 MAb for binding to the purified antigen was demonstrated in sera of tumor‐bearing and immune rats. Thus, this high‐molecular‐weight glycoprotein expressed both auto‐antigenic, tumor‐associated and tissue‐type‐restricted epitopes. The 10B12 MAb homogeneously stained the majority of colorectal carcinomas and the antigenic determinant was expressed on the cell surface of 12/13 tissue‐cultured colon‐carcinoma clones. Modulation of the cell‐surface antigen phenotype by recombinant rat interferon‐γ markedly increased expression of this determinant.
Differential utilization of the SV40 enhancer and early promoter in various human lung carcinoma cell lines
Authors: Wong Yuk‐Chor , Bernal Samuel D.
Published: February 2007

Cell lines derived from the major morphological types of human lung cancer were tested for their ability to utilize the SV40 enhancer/early promoter. These cell lines were transfected with a recombinant plasmid containing a reporter gene, coding for chloramphenicol acetyltransferase (CAT), under the control of the SV40 enhancer/early promoter. The transfected cells were then assayed for CAT activity. Non‐small‐cell carcinomas, especially squamous carcinomas, were found to be 2–3 orders of magnitude more efficient in utilizing the SV40 enhancer/early promoter than small‐cell carcinomas. The presence of different SV40 enhancer/early promoter specific DNA‐binding nuclear factors in squamous and small‐cell carcinomas was demonstrated in gel mobility shift experiments. These observations seem to suggest that the set of transcriptional regulatory factors associated with squamous carcinomas may be distinct from that associated with small‐cell carcinomas.
Reversal of multi‐drug resistance in human KB cell lines by structural analogs of verapamil
Authors: Pirker Robert , Keilhauer Gerhard , Raschack Manfred , Lechner Christina , Ludwig Heinz
Published: February 2007

Several structural analogs of verapamil were studied for their ability to reverse multi‐drug resistance (MDR) in human KB cell lines. D595, D792 and verapamil completely reversed resistance to colchicine and adriamycin. D595 and D792 had a higher reversing potency than verapamil. Devapamil, gallopamil, emopamil and D528 partially reversed MDR. The reversing potency of a drug did not correlate with its calcium antagonistic activity. No differences in reversing potency between (R)‐isomers, (L)‐isomers and the racemic forms were observed in the case of both verapamil and emopamil. (R)‐verapamil, which has less calcium antagonistic activity and less in vivo toxicity than racemic verapamil, and D792, which has higher reversing potency and less in vivo toxicity than racemic verapamil, should be suitable for clinical applications to overcome drug resistance in cancer patients.
Phenotypic characterization, karyotype analysis and in vitro tamoxifen sensitivity of new ER‐negative vulvar carcinoma cell lines, UM‐SCV‐1A and UM‐SCV‐1B
Authors: Grénman Seija E. , Van Dyke Daniel L. , Worsham Maria J. , England Barry , McClatchey Kenneth D. , Hopkins Michael , Babu V. Ramesh , Grénman Reidar , Carey Thomas E.
Published: February 2007

Two cell lines (University of Michigan squamous carcinoma of the vulva UM‐SCV‐1A and UM‐SCV‐1B) were established from the primary tumor and a malignant pleural effusion of a 62‐year‐old woman. Both tumor specimens grew vigorously in vitro and could be passaged after only 14 and 10 days in culture, respectively. Both cell lines undergo 3 population doublings in 4 days, reaching saturation densities of 5 × 105 cells/cm2, and have been carried through more than 30 in vitro passages. In nude mice the cultured cells initially formed tumors but these regressed 2–3 weeks after inoculation. The regressing mouse tumors consisted of poorly differentiated squamous carcinoma surrounded by an inflammatory lymphoid infiltrate. The UM‐SCV‐1 cell lines express membrane antigens typically displayed by squamous‐cell carcinomas. These include the HLA class‐1 light chain β2‐microglobulin, pemphigus, pemphigoid, and the α6β4 integrin defined by the UM‐A9 monoclonal antibody (MAb). In contrast to the A431 vulvar carcinoma, these tumor lines do not have amplified expression of the epidermal growth factor (EGF) receptor. Although tissue from the primary tumor contained low levels of estrogen receptor activity, no receptor activity was detected in the cell lines. Nevertheless, both lines were sensitive to growth inhibition by tamoxifen. This effect was not reversible by estradiol, indicating an estrogen‐receptor‐independent mechanism. The tumors were both hypotetraploid, contained the same chromosome rearrangements and had stable karyotypes in vitro. Each contained inv(1)(p36.3q32.1), del(4)(q12), dic(4;11)(q12;p11.2), i(5p), der(6)t(3;6)(q25.1;p21.1), several rearrangements involving chromosomes 8 and 14, +i(13), i(18p), a dicentric t(11;19), and 2 or 3 unidentified markers. Since the karyotypes of both tumors were the same, no major karyotypic change was associated with metastatic spread. These paired primary and metastatic SCC lines from an unusually aggressive vulvar carcinoma provide an in vitro model for analysis of the biological basis of this tumor's behavior.
Establishment and characterization of a leukemic murine cell line derived from MCF 247 MuLV‐induced T‐cell lymphoma
Authors: Dolcetti Riccardo , Carbone Antonino , Barlati Sergio , Maestro Roberta , Rizzo Silvana , Sonego Fulvia , Feriotto Giordana , Bellotti Daniela , Boiocchi Mauro
Published: February 2007

Understanding of the leukemic evolution of human non‐Hodgkin's lymphomas is hindered by the lack of appropriate animal models. For this purpose, a highly leukemic cell line NQ22, derived from a MCF 247 murine leukemia virus (MuLV)‐induced murine T‐cell lymphoma, was established, and its preliminary characterization is described. The NQ22 cell line is easily transplantable subcutaneously (s.c.) into syngeneic AKR mice exhibiting early peripheral blood invasion and widespread dissemination with a leukemic pattern of infiltration. Such peculiar in vivo behavior is a stable phenotypic feature, probably determined genetically. Biological and differentiation characteristics of the NQ22 cell line were analyzed and compared to those of other non‐leukemic T‐lymphoma lines. In addition, no evidence of possible involvement of plasminogen activator (PA) enzymes and of their inhibitors (PA1) in the spreading ability of NQ22 cells was observed.
The CMF‐regimen. Modulation of cyclophosphamide uptake and clearance by methotrexate and fluorouracil
Authors: De Bruijn Ernst A. , Geng Yi , Hermans Jo , Driessen Oscar
Published: February 2007

Influence of the 2 antimetabolites used in the CMF‐regimen, methotrexate (MTX, M) and fluorouracil (FUra, F) on in vivo pharmacokinetics of orally administered cyclophosphamide (CY, C), were studied in WAG/Rij rats. Blood plasma concentrations of CY following oral administration were monitored in single‐agent CY, in CY + MTX (CM), in CY + FUra (CF) and in CY + MTX + FUra (CMF) treatments. Each treatment group consisted of at least 10 rats. CY was determined in 50 μl of plasma by capillary gas chromatography on the first day of chemotherapy. Statistical analysis of blood plasma concentration data revealed a significant influence of both MTX and FUra on CY input/output function (p: 0.01). MTX and FUra significantly increased the area under the plasma concentration time‐curve, whereas tmax was significantly prolonged in CF and CMF treatment groups (p: 0.01). It is suggested that MTX and FUra interact at the site of CY pre‐systemic metabolism, including first‐pass metabolism, subsequently resulting in prolonged absorption.