bluuebunny/crossref_metadata_2025 huggingface dataset:
Actin filaments and tumorigenicity in a fischer rat embryo fibroblast cell line (3Y1) transformed by ultraviolet‐irradiated HSV
Authors: Tsuchie H. , Katsumoto T. , Hama S. , Hattori N. , Morimoto H. , Kamahora T. , Kurimura T.
Published: February 2007

The characteristics of the cytoskeleton of a Fischer rat embryo fibroblast cell line (3Y1) transformed by ultraviolet (UV)‐irradiated HSV were studied by indirect immunofluorescence using anti‐actin IgG. Parental 3Y1 cells possessed well‐developed actin filaments, while 3Y1 cells transformed by UV‐irradiated HSV also retained well‐developed actin filaments. Transformed cells were divided into 2 groups according to tumorigenicity in newborn Fischer rats; one had a strongly tumorigenic potential and the other a weakly tumorigenic potential. Tumor‐derived cell lines exhibited a highly tumorigenic potential, and were also divided into 2 groups, one with well‐developed actin filaments and the other without well‐developed actin filaments. Our results suggested that transformation or tumor formation by HSV is a multi‐step process and that morphological loss of actin filaments in the cells is not essential to the tumorigenic potential of the cells transformed by HSV.
Altered growth properties and cell surface changes in ras transformed mouse bladder epithelium
Authors: Summerhayes I. C. , Malone P. , Visvanathan K.
Published: February 2007

Transfection of the c‐Ha‐ras‐1 oncogene, cloned from EJ/T24 cells, into different mouse bladder epithelial cell lines resulted in the acquisition of tumorigenic potential and, in all but one cell line (MB331), anchorage‐independent growth. Sera from syngeneic mice bearing tumours immunoprecipitated an 18 kDa protein from ras‐transfected urothelial cells which was not detectable in their parental counterparts. Screening of a limited panel of mouse cell lines showed this protein to be urothelium‐specific and associated with the expression of an activated ras gene. Polyoma middle T and v‐myc‐transfected bladder epithelial cells did not express this 18kDa protein. Localization of this protein to the cell surface was demonstrated by immunofluorescence and absorption studies.
Production and characterization of monoclonal antibodies against human neuroblastoma
Authors: Schönmann S. M. , Iyer J. , Laeng H. , Gerber H. A. , Käser H. , Blaser K.
Published: February 2007

MAb were derived from mice immunized with cells of the human neuroblastoma line IMR‐32. Five hybridomas were selected according to their selective binding to human cell lines, tumors and normal tissues. One of them, CE7, reacted with all sympatho‐adreno‐medullary cells (neuroblastoma, ganglioneuroblastoma, ganglioneuroma, pheochromocytoma, adrenal medulla, sympathetic ganglion cells). Weak cross‐reactivities were observed with melanocytes and with some human melanoma and glioma cell lines. The antigen recognized by CE7 was markedly expressed on neuroblastoma tumors of all histological grades, independently of the adrenergic or cholinergic nature of these cells. MAb derived from clones AD2, BCI, BC4 and CB10 bound variably to some, but not to all, neuroblastoma cells. By using these MAb, 3 phenotypes of neuroblastoma lines could be distinguished. The binding profiles of these types, however, showed no correlation with origin of the cell lines or stage of the disease.
Transmission of human T‐cell leukemia virus (HTLV‐I) by blood transfusion: Demonstration of proviral DNA in recipients' blood lymphocytes
Authors: Sato Hiroyuki , Okochi Kazuo
Published: February 2007

Human T‐cell clones bearing antigens encoded by human T‐cell leukemia/lymphoma virus (HTLV‐I) were isolated from 6 patients who produced antibodies against HTLV‐I after having received anti‐HTLV‐I‐positive blood units containing cell components. On the other hand, it was not possible to isolate clonal cells carrying viral antigens from the recipients who did not produce antibodies. The clonal cell lines had the same surface markers as neoplastic cells of adult T‐cell leukemia and had the HLA phenotype of the recipients themselves. Proviral DNA of HTLV‐I was demonstrated in each of the clonal cell lines. The site of integration was different in each case even if the clones were derived from the same recipient. These results indicate that blood transfusion can cause persistent HTLV‐I infection.
The role of prostaglandin E1 in ornithine decarboxylase induction by tumor promoters
Authors: Ordman Alfred B. , Simsiman R. C. , Cleaveland Jeffrey S. , Boutwell R. K.
Published: February 2007

The effect of topical application of PGE on induction of ODC in mouse epidermis was measured. When direct induction of ODC by TPA was blocked by also applying indomethacin, maximum ODC activity occurred only when PGE was applied simultaneously with TPA 41/2 hr before killing of the mice. If either TPA or PGE was applied at other times, ODC activity decreased substantially. Induction of ODC by mezerein was blocked by indomethacin but restored by PGE, as was observed with TPA, but induction by ethyl phenylpropiolate was not affected by indomethacin or PGE. DMBA did not cause a consistent increase in ODC activity, nor was its inductive action affected by indomethacin or PGE. However, another weak inducer, acetic acid, exhibited elevated ODC activity when PGE was also applied. Inhibition by topical retinoic acid of ODC induction by TPA was partially overcome in a dose‐response fashion by PGE. The results indicate that at least 2 events, elevation of PGE and another independent event, are required for induction of ODC activity. It appears that TPA causes at least 4 independent events essential for tumor promotion. A model for the events in the 2‐stage tumor promotion model is proposed.
Presence of human papillomavirus type‐16 and type‐18 DNA sequences and their expression in cervical cancers and cell lines from japanese patients
Authors: Tsunokawa Youko , Takebe Naoko , Nozawa Shiro , Kasamatsu Tatsuhiro , Gissmann Lutz , Hausen Harald Zur , Terada Masaaki , Sugimura Takashi
Published: February 2007

Southern blot analyses of surgical specimens of cervical carcinoma from Japanese patients showed that 3/9 samples contained human papillomavirus (HPV) type‐16 DNA sequences, and 2 contained HPV type‐18 DNA sequences. By Northern blot analyses, RNA transcripts of HPV DNA sequences were demon‐trated in some of the tissues containing HPV type‐16 or HPV type‐18 DNA sequences. Two cell lines established from cervical cancers of Japanese patients also contained HPV type‐18 genomes and these cell lines contained HPV type‐18 transcripts. Two other cervical cancer cell lines from a Japanese patient were found to contain HPV type‐16 DNA sequences and their RNA transcripts.
Virus production and surface marker expression in human lymphocytes immortalized following dual infection with human T‐cell leukemia virus and Epstein‐Barr virus
Authors: Mochizuki Shigenobu , Kawamura Kiyoko , Maruyama Koshi
Published: February 2007

By co‐cultivating peripheral blood lymphocytes from normal male donors with cells from a line designated Lma‐66VP, established from a female donor and simultaneously producing both HTLV‐I and EBV, 2 continuous culture lines were obtained. Normal male lymphocytes were considered to be immortalized by co‐cultivation because they were of male karyotype and lacked the 3q + chromosome that was observed in all Lma‐66VP cells. These immortalized cultures were designated Co‐culture I and Co‐culture 2. After prolonged cultivation, chromosome abnormalities characteristic of each co‐culture line were observed in all cells examined, indicating their clonal origin. Immortalized cells initially produced both HTLV‐I and EBV. Although production of EBV was not seen in the co‐culture lines after prolonged cultivation when all cells examined had the characteristic chromosomal abnormality indicating their clonality, EBNA+ cells persisted. Thus, clonal cells were doubly infected by HTLV‐I and EBV. While the majority of clonal cells in Lma‐66VP, Co‐culture I, and Co‐culture 2 lines expressed a surface marker of B‐cells, small numbers of cells expressed a T‐cell surface marker. These findings demonstrate that human lymphocytes immortalized following dual infection by HTLV‐I and EBV exhibit chromosome rearrangement, resulting in expression of surface markers inconsistent with their differentiation lineage.
Modulation of protein kinase C and Ca2+ lipid‐independent protein kinase in lymphoma induced by moloney murine leukemia virus in BALB/c mice
Authors: Wolfson M. , Aboud M. , Ofir R. , Weinstein Y. , Segal S.
Published: February 2007

We investigated the possible role of protein kinase C (PKC) in the progression of Moloney murine leukemia virus (Mo‐MuLV)‐induced lymphoma in BALB/c mice. Mice injected with Mo‐MuLV on the first day after birth developed lymphoma within 1 1/2 3 months. The development of lymphoma was characterized by a gradual increase in the number of spleen cells. However, no analogous changes could be detected in the thymuses of these mice, although cells of both organs were found to be virus producers as early as 3‐4 weeks after inoculation. PKC activity, which was assayed in extracts of spleen and thymus cells, declined gradually during the development of lymphoma. Concomitantly with this decline, a progressive appearance of Ca2+/ lipid‐independent protein kinase activity was observed. TPA treatment of intact cells from normal mice reduced the level of soluble PKC activity, while inducing Ca2+/lipid‐independent phosphorylation. By contrast, TPA had no effect on these enzymatic activities in cells derived from leukemic mice. Spleen enlargement caused by injection of a non‐leukemogenic inflammatory agent such as mineral oil was ineffective in this respect, suggesting that the PKC‐Ca2+ /lipid‐independent protein kinase modulation is associated with the virally induced leukemogenesis.
Phenotypes of placental‐type alkaline phosphatase in seminoma sera as defined by monoclonal antibodies
Authors: Wahren B. , Hinkula J. , Stigbrand T. , Jeppsson A. , Andersson L. , Esposti P. L. , Edsmyr F. , Millán J. L.
Published: February 2007

PLAP‐like enzymes could be detected in serum of patients with primary testicular tumors, in particular seminomas. The use of a panel of monoclonal antibodies (MAbs) permitted typing into 6 different testicular serum phenotypes, of which one appeared similar to a placental type (II) and 2 have not been previously described. Most tumor sera belonged to type I, as described for seminoma tissues. With a more advanced tumor the mean serum PLAP‐like levels increased. After operation, after radiotherapy or with no evidence of disease lower or non‐detectable enzyme levels were found. In typing the tissue PLAP‐like antigen in serum of the same patient or sera from recurrences of a seminoma in the same patient, the same phenotypes of PLAP‐like antigen were usually but not always found. None of the 6 phenotypes appeared to confer a poorer prognosis. We conclude that the expression of PLAP‐like antigen is eutopic and is enhanced by testicular malignancy, especially in seminoma cells.
Mode of transmission of human T‐cell leukemia virus type I (HTLV I) in a human promyelocytic leukemia HL60 cell
Authors: Hiramatsu K. , Masuda M. , Yoshikura H.
Published: February 2007

HTLV‐I propagated in IMR90 human diploid fibroblasts was transmitted to human myeloid leukemia HL60 cells at a low efficiency. After co‐cultivation for 3 months, the viral genome was detected in 14/48 HL60 cell clones. Among the 14 HTLV‐I‐infected clones, 8 contained subgenomic fragments alone or in addition to the complete HTLV‐I genome. The frequency of deleted proviruses (9/24 total proviruses) was unexpectedly high. Hirt's supernatant of some of the clones harboring complete HTLV‐I genome(s) in the chromosome contained both linear and circular HTLV‐I proviral DNAs. The circular DNAs were composed of one LTR and 2 LTR closed circular proviruses. These clones produced infectious HTLV‐I constitutively, which was proved by transmission of the viral genome into fresh IMR90 cells by co‐cultivation. However, in these clones, re‐integration of extrachromosomal provirus into their own chromosomes was not observed.
Potential carcinogenicity of Aza‐aromatic hydrocarbons: Azabenz(A)anthracenes
Authors: Miao R. M. , Tanga M. J. , Suk W. A. , Miller A. , Reist E. J.
Published: February 2007

The potential carcinogenicity of 3 azabenz(a)‐anthracenes was determined in vitro. The 3 compounds tested were 1‐, 2‐, and 9‐azabenz(a)‐anthracene. The initial assay was chemical carcinogen‐induced enhancement of anchorage‐independent survival of Rauscher leukemia virus‐infected Fischer rat embryo cells, 2FR450 (2FR4). Cells treated with 2‐ and 9‐azabenz(a)anthracene showed dose‐dependent increased survival. After continued subculturing, the surviving cells from 2‐ and 9‐azabenz(a)anthracenetreated cultures displayed morphological transformation and ability to grow in semi‐solid medium. Mocktreated controls and I‐azabenz(a)anthracene‐treated cultures did not show either of these properties. These data suggest that certain azabenz(a)anthracenes are potential carcinogens.
Fecal bile acids and neutral sterols in rats with spontaneous colon cancer
Authors: Hayashi Eijiro , Amuro Yoshiki , Endo Tomoichiro , Yamamoto Hideo , Miyamoto Makoto , Kishimoto Susumu
Published: February 2007

Fecal bile acids and neutral sterols of spontaneous colon cancer‐bearing Wistar‐Furth strain rats were examined and compared with those of control rats of the same strain by gas chromatography and gas chromatography‐mass spectrometry. The amount of total fecal bile acids and the percentage of fecal lithocholic acid were almost similar in both groups, but the percentage of fecal deoxycholic acid was significantly higher in the colon cancer group than in the controls. The amount of total fecal neutral sterols and the percentage of fecal coprostanol were also markedly higher in the colon cancer group than in the controls. The percentage of cholesterol and the amount of β‐sitosterol were, however, almost similar in the two groups. These results indicate that fecal deoxycholic acid and fecal coprostanol are notably higher in rats with colon cancer than in controls.
A radioimmunoassay for the detection of a human tumor‐associated glycoprotein (tag‐72) using monoclonal antibody B72.3
Authors: Paterson Andrew J. , Schlom Jeffrey , Sears Henry F. , Bennett Jeffrey , Colcher David
Published: February 2007

Monoclonal antibody (MAb) B72.3 was generated against a carcinoma metastasis and has been shown to bind with a high degree of selectivity to a tumor‐associated glycoprotein (TAG‐72) found in human colon and breast carcinomas, while showing minimal reactivity to any normal adult tissues. Competition radioimmunoassays have been developed for the detection of TAG‐72 in tumors and sera from both athymic mice bearing human carcinoma xenografts and patients with colon, breast and other carcinomas. The distribution of TAG‐72 in human tumor xenografts was restricted to tumors originating from the LS‐174T human colon carcinoma, with no significant reactivity being detected in human tumor xenografts from a different colon carcinoma, a human breast carcinoma, or a human melanoma. The levels of TAG‐72 in clinical material obtained from surgery were examined; high levels were found in colon carcinomas and to a lesser extent in breast carcinomas, while no detectable levels were found in normal tissues. Sera from apparently normal patients contained a mean level of 2.2 units per ml of TAG‐72. A level of 3 standard deviations above the mean level of TAG‐72 found in normals was employed as a cut‐off value to indicate a positive test result in subsequent studies. No patient with inflammatory disease or other benign colon disease exhibited elevated levels of TAG‐72. Seven out of 20 advanced colon cancer patients and 7 out of 20 patients with other carcinomas had elevated serum levels of TAG‐72. The serum TAG‐72 levels were compared with the serum levels of antigens recognized by the MAbs currently used to screen sera of carcinoma patients (CEA, GICA, and OC125). It was clearly demonstrated that TAG‐72 is different from these antigens and can be found in some sera in which no antigen is detected by otherwise available MAb RIAs.
Immunohistologic expression of blood‐group antigens in normal human gastrointestinal tract and colonic carcinoma
Authors: Cordon‐Cardo Carlos , Lloyd Kenneth O. , Sakamoto Junichi , McGroarty Mary Ellen , Old Lloyd J. , Melamed Myron R.
Published: February 2007

A panel of 7 mouse monoclonal antibodies and the lectin from Ulex europeus, detecting blood‐group‐related antigens of the ABH and Lewis systems, have been used to define the distribution of these antigenic structures within the human gastrointestinal tract, and to characterize their expression and modulation in colorectal carcinomas. The reagents employed detect the following blood‐group specificities: A (all variants), B. H (type 2), Lewisa, Lewisb, × (Lewisx), Y (Lewisy) and type I precursor chain. Immunohistochemical studies demonstrate that these antigens are differentially expressed in various cell types and developmental stages of the human gastrointestinal tract. ABH expression undergoes developmental modulation in the human colorectal tract from positive to negative during embryogenesis, and is lost in adult cells. Colorectal tumors exhibit neosynthesis of ABH specificities that appear in tumor cells, and accumulation of the precursor antigens. They also show increased expression of Lewis antigens, especially Y determinant, which has a restricted pattern of distribution in normal tissues and is not found in normal colonic mucosa. Enhancement of the Lewis antigens is observed in all colorectal tumors analyzed, regardless of blood‐group type and secretory status of the individuals studied. Tumor modulation of these antigens may be related to activation of suppressed genes and enhancement of fucosyltransferases.
Human‐human hybridomas for the study of anti‐tumor immune response in patients with colorectal cancer
Authors: Borup‐Christensen P. , Erb Karin , Jensenius J. C. , Nielsen B. , Svehag S.‐E.
Published: February 2007

Human‐human hybridoma technology was evaluated for the study of humoral immune reactions of colorectal cancer patients against their own tumors. Six fusions were carried out with lymphocytes from mesenteric lymph nodes from patients with colorectal cancer, using the human B‐lymphoma cell line LICR‐LON‐HMy‐2 as fusion partner. A total of 294 wells with cell growth were obtained. Supernatants from 26 of these reacted in enzyme‐linked immuno‐sorbent assay (ELISA) with one or more colon cancer cell lines. Cells from only one of these wells (D 4213) could be cloned. The clone was shown to produce antibody which by immunocytochemical analysis reacted with a panel of colon cancer cell lines and melanoma cell lines but not with several other cancer cell lines or normal human leukocytes. By immunohistochemical analysis on formalin‐fixed paraffin‐embedded tissue this antibody reacted strongly with antigen expressed by autologous and allogeneic colorectal cancers. Faint staining could occasionally be observed on normal colon epithelium. D4213 is a hybrid cell line since it is setraploid and produces kappa and lambda light chains as well as gamma and mu chains, whereas HMy‐2 produces only kappa and gamma chains. The study suggests that patients may possess B cells producing antibody reactive with their own malignant cells.
Physicochemical purification and immunological characteristics of ductal carcinoma antigen
Authors: Papsidero Lawrence D. , Johnson Edward A.
Published: February 2007

Ductal carcinoma antigen (DCA), as recognized by monoclonal antibody (MAb) F36/22, was purified in high yield and to homogeneity from malignant effusions by means of physicochemical techniques. Fractionation procedures were monitored by immunoenzymometric assays. Preparations purified over 2,000‐fold were obtained through acid‐extraction, lectin‐chromatographies and gel filtration steps. Purified antigen demonstrated a single component upon electrophoretic examination. This material had a high molecular weight and high immunoreactivity. Neuaminidase treatments failed to perturb the antigenicity of the component, indicating an absence of sialic acid residues at the antibody‐combining site. Further, extensive proteolytic digestion effected the release of heterogeneous glycopeptides which retained immunologic activity, suggesting the presence of carbohydrate at the active site. Immunologically, DCA was compared with other tumor‐associated mucin‐like gly‐coproteins which have been detected in the circulation of patients. Results indicated no cross‐reactivity between DCA and other mucins, including CA 19–9 and CA‐125. Further, no antigenic relationship was noted for purified carcinoembryonic antigen (CEA), also a heavily‐glycosylated structure. These data thus suggest that immunological monitoring of disease may be approached using a selected panel of antigenically distinct reagents.
Biochemical regulation of adenylate cyclase in murine melanoma clones with different metastatic properties
Authors: Sheppard J. R. , Lester Bruce , Doll John , Buscarino Charles , Gonzales Elisa , Corwin Steven , Greig Russell , Poste George
Published: February 2007

The regulation of adenylate cyclase in murine melanoma tumor cell clones with different metastatic capacities has been studied in intact cells and isolated membrane preparations. Analysis of the responses of intact cells from a number of B16 melanoma clones revealed that treatment with melanocyte‐stimulating hormone (MSH) or the diterpene, forskolin, produced significantly greater accumulation of intracellular cyclic adenosine 3′,5′ monophosphate (cAMP) in strongly metastatic clones than in weakly metastatic tumor cell clones. In contrast, in isolated membranes from the same panel of clones, the extent of activation by forskolin but not by MSH correlated with metastatic capacity. Sodium fluoride and 5′‐guanyl‐ß‐γ‐imidodiphosphate [Gpp(NH)p] also stimulated adenylate cyclase in isolated membranes but the extent of activation did not correlate with the metastatic behavior of the donor cells. A combination of forskolin and Gpp(NH)p proved to be a sensitive prospective indicator for identifying differences in the metastatic capabilities of individual B16 melanoma clones. Adenylate cyclase in membrane preparations from strongly metastatic B16 clones displayed synergistic activation but stimulation of the enzyme from weakly metastatic clones was less than additive. To test the generality of these findings, similar investigations were performed on B16‐BL6 melanoma cells, a highly invasive subline of the B16 melanoma, and the K1735, an ultraviolet‐light‐induced murine melanoma arising in a different mouse strain (C3H). Consistent with their high metastatic potential, clones derived from the B16‐BL6 melanoma displayed elevated levels of hormon‐ally‐stimulated adenylate cyclase, thereby confirming, for this tumor system, a close association between hormonal responsiveness and metastatic capacity. In contrast, K1735 melanoma cell clones exhibited significant interclonal variation in adenylate cyclase activity and metastatic performance, but no consistent relationship between the two traits was detected. Differences in the regulation and/or the intrinsic catalytic capacity of adenylate cyclase may account, at least in part, for the variation in hormonal responsiveness observed among B16 clones with distinct metastatic properties and suggest that cAMP‐dependent molecular processes may be required for the expression of B16 melanoma experimental metastatic potential. The failure to detect a similar association between cAMP metabolism and metastatic performance in the K1735 melanoma cells indicates that this molecular process is not an invariant feature of metastatic tumor cells and suggests that metastatic competence in tumor cells arising from a common cell lineage involves diverse phenotypic aberrations.
Characterization of a tumorigenic murine T‐lymphoid‐cell line spontaneously derived from an IL‐2‐dependent T‐cell line
Authors: Mann Varda , Szyf Moshe , Razin Aharon , Chriqui‐Zeira Evelyne , Kedar Eli
Published: February 2007

The establishment of IL‐2‐independent T‐cell lines spontaneously derived from long‐term IL‐2‐dependent cytotoxic T‐cell lines is described. Two lines (cloned and uncloned) studied in detail have shown the following characteristics: (1) Permanent loss of IL‐2 dependence. (2) Partial or complete loss of both cytotoxic activity and the IL‐2 receptor. (3) Increased expression of T‐cell membrane markers (Thy 1.2, Lyt 1.2) compared with the parental line. (4) Lower level of DNA methylation than in freshly obtained lymphoid cells. (5) Different karyotypic pattern from the parental IL‐2‐dependent line, with a mean number of 39–40 chromosomes and a resemblance to T leukemic lines. (6) Leukemia caused in normal syngeneic C57BL/6 mice by the uncloned line, in contrast to the cloned IL‐2‐independent line or the parental dependent line. Unlike established leukemic lines, however, the independent line gave rise to tumors which regressed in some mice within a few days of their appearance. These findings suggest that T‐cell lines maintained with IL‐2 for prolonged periods of time (> 3 months) can undergo transformation and, therefore, should not be utilized for immunotherapeutic purposes.
Separation, phenotyping and limiting dilution analysis of T‐lymphocytes infiltrating human solid tumors
Authors: Whiteside Theresa L. , Miescher Sylvia , Hurlimann Jean , Moretta Lorenzo , Fliedner Vladimir Von
Published: February 2007

Tumor‐infiltrating lymphocytes (TIL) were obtained by a combination of mechanical release and enzymatic disaggregation from 35 human solid tumors. The number of lymphocytes in TIL‐enriched suspensions varied from 1 × 104to 7.6 × 106per wet gram of tumor. The TIL preparations separated by differential centrifugation on Ficoll‐Hypaque gradients contained 10–95% of TII+cells (mean 50%), and tumor cells accounted for the other major cellular component. Macrophages, NK cells, B cells and granulocytes were infrequently seen. Morphologically, TIL‐T were small non‐activated cells. They expressed the TII and T3 antigens but not the receptor for IL‐2 (IL‐2R) or HLA‐DR antigens as determined by double immunofluorescence staining. Rare TII+/IL‐2R+cells were recovered only from colon and lung carcinomas. The mean T4/T8 ratio in 12 TIL preparations was 1.1 ± 0.8. Immunohistology with monoclonal antibodies (MAbs) performed in 31/35 tumors confirmed that the TII+cells infiltrating solid tumors rarely expressed the IL‐2R and that the cell content of suspensions enriched in TIL was comparable to that determinedin situ. The recovered TIL were cloned in a microculture system that permits proliferation of nearly all normal peripheral blood T lymphocytes (PBL‐T). Under these culture conditions, frequencies of the proliferating T lymphocyte precursors (PTL‐P) were depressed in both the TIL preparations (< 0.01 to 0.39) and patients' PBL‐T (0.05 to 0.5). These low frequencies of PTL‐P were seen in patients with all tumor types, both primary and metastatic.
Influence of growth phase, nutrition and hypoxia on heterogeneity of cellular buoyant densities in in vitro tumor model systems
Authors: Luk Catherine K. , Sutherland Robert M.
Published: February 2007

EMT6/Ro multicellular spheroids were sequentially dissociated into outer, middle and inner layers, and the buoyant densities of these subpopulations were compared to aerobic or exponential or plateau‐phase monolayers by isopycnic density centrifugation using Percoll as the gradient medium. Spheroids were more heterogeneous in buoyant density distributions than monolayer cultures. The outer‐layer cells of spheroids and fed plateau monolayers peaked at a similar density; the majority of middle‐layer cells banded close to unfed plateau and exponential hypoxic monolayers, while most inner‐layer cells banded where fed or unfed plateau hypoxic monolayer peaked in density. Our data suggest that metabolic heterogeneity due to growth phase, together with altered cellular microenvironment brought about by differences in oxygenation and external nutrient supplies between spheroids and monolayers, can be manifested in one biophysical parameter such as buoyant density.
Ca2+ and calmodulin are involved in the processes conferring stability to DNA in proliferating neoplastic cells
Authors: Lönn Ulf , Lönn Sigrid
Published: February 2007

We have examined, in proliferating neoplastic cells, the effect on DNA of EGTA (a chelator of Ca2+) and W7 (an inhibitor of calmodulin). The treatment results in release of single‐stranded DNA fragments (2‐10 kb) from pre‐labelled HMW DNA. When DNA from synchronized neoplastic cells in S phase is examined, almost all pre‐labelled DNA appears as short fragments. However, fragmentation does not occur in growtharrested cells or in normal cells. Furthermore, fragmentation can be prevented by incubating cells in excessive amounts of Ca2+. Hence Ca2+ and the Ca2+‐binding protein calmodulin are involved in the processes conferring stability to DNA in proliferating neoplastic cells.
Biochemical characterization and biologic activities of 82‐and 86‐kDa tumor antigens isolated from a methylcholanthrene‐induced sarcoma, CII‐7
Authors: Dubois Garrett C. , Law Lloyd W.
Published: February 2007

Two tumor‐specific antigens, with molecular weights of approximately 82 and 86 kDa, have been isolated and purified to apparent homogeneity from the methylcholanthrene‐induced sarcoma, CII‐7. The method of purification used was essentially that previously employed for the isolation of the 82‐ and 86‐kDa antigens from the Meth A sarcoma and the 86‐kDa antigen from mKSA sarcoma. Cytosolic fractions were subjected to hexylamine agarose chromatography, Sepharose S‐300 filtration and hydroxylapatite chromatography. A final step, HPLC‐DEAE chromatography, was necessary for the purification of the 82‐kDa protein. Both isolated antigens retained their specific immunogenicity for CII‐7 as determined by in vivo tumor rejection assays, and failed to influence the growth of other syngeneic sarcomas, CI‐4, CII‐10 and mKSA, which have their own unique TATA. The 82‐and 86‐kDa antigens appear to be distinct proteins that are well conserved in nature and may represent members of distinct families of TATA.
Trisomy and tetrasomy for long arm of chromosome 1 in near‐diploid human endometrial adenocarcinomas
Authors: Couturier J. , Vielh P. H. , Salmon R. , Dutrillaux B.
Published: February 2007

Karyotypic analysis by R‐banding after short‐term culture, carried out on 7 cases of human endometrial adenocarcinoma, showed in 4 of these a trisomy or a tetrasomy for the long arm of chromosome I. In the 4 cases, these imbalances were due to rearrangements involving centromeric or para‐centromeric breakpoints: 46, XX,−16, + der(1q16p) t(1;16)(1p16q;1q16p); 46, XX,−21, + der(1q21q) t(1;21)(1p21p;1q21q);46, XX,−21, + der(21) t(1;21)(q11;p13); 48, XX, +2, + i(1q). Two other cases showed only a numerical aberration: 47, XX, + 10 and 47, XX, + 12. In the last case, only cells with apparently normal karyotype were seen. In the 4 cases with an anomaly of chromosome I, two normal I chromosomes coexisted with abnormal elements. This shows that the rearrangement very likely occurred in G2 phase of the cell cycle.
Incidence of soft‐tissue cancers in blacks and whites in New York State
Authors: Polednak Anthony P.
Published: February 2007

Incidence rates for several histologic types of soft‐tissue cancers were compared between black and white residents of New York State during 1976–81, using data from the population‐based New York State Cancer Registry. Incidence rates for leiomyosarcoma, neurofibrosarcoma and malignant neurolemmoma were higher for blacks than for whites in almost all age groups. Black‐white differences in malignant mesenchymoma, while not statistically significant, were similar to those from other population‐based studies. There was no evidence for black‐white differences in certain other histologic types (e.g., blood‐vessel cancers, lymphangiosarcoma). Black‐white differences in leiomyosarcoma included higher rates for the uterus in black than in white females, and higher rates for the stomach in blacks of both sexes. Possible explanations for these differences in soft‐tissue cancer incidence were discussed, along with comparisons with limited data on black populations in Africa.
Measurement of the survival of human tumor cells after implantation in athymic nude mice
Authors: Romijn J. C. , Verkoelen C. F. , Schroeder F. H.
Published: February 2007

A method has been developed to determine the survival of human tumor cells after implantation in athymic nude mice. The method involves the labelling of cells of the prostatic cancer cell line PC‐3 with tritiated thymidine after attachment of the cells to Millipore filter disks. The amount of radioactive label incorporated in the cells remained constant for at least 48 hr during in vitro incubation. After s.c. implantation of the filter disks in nude mice, however, the fraction of surviving cells, reflected by the amount of radioactivity persisting on the filters, rapidly decreased with time. Only a relatively small fraction (approx. 10%) of the cells survived for more than 24 hr under these conditions. Pretreatment of the mice with cyclophosphamide (100 mg/kg, 24 hr prior to implantation) resulted in a 2‐fold increase of tumor‐cell survival. Likewise, the frequency of tumor takes after injection of PC‐3 cells was enhanced following cyclophosphamide pretreatment. These results indicate that immune defense mechanisms might be partly (but not completely) responsible for tumor‐cell rejection in nude mice. The technique described here might be a valuable tool for the rapid evaluation of possible effects of various types of pretreatment of the host animal on tumor cell survival and, thereby, on tumor growth.
Quantification and characterization of high‐affinity membrane receptors for tumor necrosis factor on human leukemic cell lines
Authors: Scheurich Peter , Ücer Ugur , Krönke Martin , Pfizenmaier Klaus
Published: February 2007

The expression of specific membrane receptors for TNF‐aipha was determined on various human leukemic cell lines differing in their sensitivity to the growth‐inhibitory activity of TNF‐alpha. Binding studies with 125Mabelled TNF‐alpha indicated specific binding in 8/10 cell lines with approximately 10‐fold differences in the quantity of TNF‐alpha bound by these distinct cell lines. Scatchard analyses of TNF‐binding revealed the existence of high‐affinity membrane receptors (Kd 1.5 × 10−10 M) and approximately 3,000 binding sites/cell on both U937 and K562, representing 2 cell lines with high and low TNF sensitivity, respectively. Disuccin‐imidyl‐suberate cross‐linking of receptor‐bound 125I‐TNF‐alpha and SDS‐PAGE of membrane preparations of either U937 or K562 cells suggest a single receptor protein with an apparent molecular weight of 76 kDa. Comparison of the TNF‐alpha binding capacity versus in vitro growth inhibition provides evidence that sensitivity to TNF‐alpha is determined both at the level of receptor expression and at a post‐receptor level. IFN‐gamma strongly enhanced the TNF‐alpha‐mediated growth inhibition of 3 sensitive cell lines, but had no effect on 7 other leukemic cell lines with little or no TNF sensitivity. No correlation was found between this enhancement of TNF sensitivity and the IFN‐gamma‐mediated increase in TNF‐cell membrane receptors, suggesting that IFN‐gamma predominantly exerts its synergistic effect distal to TNF‐binding.
Requirement for the continual presence of lipopolysaccharide for production of tumor necrosis factor by thioglycollate‐induced peritoneal murine macrophages
Authors: Gifford George E. , Lohmann‐Matthes Marie‐Luise
Published: February 2007

This work demonstrates the need for the continued presence of lipopolysaccharide (LPS) for tumor necrosis factor (TNF) production by thioglycollate‐induced peritoneal macrophages. Removal of LPS at any time resulted in the abrupt cessation of further TNF production. The readdition of LPS resulted in further production of TNF but the yield was limited to the amount that would have been produced had the LPS not been removed.
Resistance of line 63 chickens to reticuloendotheliosis‐virus‐induced bursa‐associated lymphomas
Authors: Fadly Aly M. , Witter Richard L.
Published: February 2007

Chickens of lines 63 and 1515 × 71 were inoculated with the chick syncytial strain of reticuloendotheliosis virus (REV) or with the Rous‐associated virus‐1 of avian leukosis virus (ALV) at hatching. At 4, 10, 16, and 36 weeks post inoculation (PI), chickens were tested for REV‐and ALV‐induced viremia and antibody. The incidence of REV‐or ALV‐induced bursa‐associated lymphomas in line 63 chickens was compared with that in line 1515 × 71 chickens. Inoculation of REV at hatching resulted in immunological tolerance to the virus in line 63 but not in line 1515 × 71 chickens. Between 70% and 100% of line 63 chickens remained viremic and lacked REV antibody throughout the experimental period of 36 weeks. In contrast, ALV‐inoculated chickens of both lines had antibody by 16 weeks PI. The frequencies of REV‐and ALV‐induced bursa‐associated lymphomas in line 63 chickens were significantly lower than in line 1515 × 71 chickens. Further, the incidence of bursa‐associated lymphomas induced by REV in line 1515 × 71 chickens was significantly lower than that induced by ALV. These results suggest that: (1) the genetic constitution of the host may influence the immunological response to REV infection; (2) chickens resistant to ALV‐induced bursa‐associated lymphomas are equally resistant to such lymphomas induced by another unrelated avian retrovirus, REV; and (3) ALV is a more potent inducer of bursa‐associated lymphomas than REV.
Differentiation antigens in fetal human pancreas. Reexpression in cancer
Authors: Escribano Maria Juana , Cordier Jocelyne , Nap Marius , Kate Fiebo J. W. Ten , Burtin Pierre
Published: February 2007

An antiserum was raised against pancreatic extracts obtained from human fetuses under 5 months of gestational age. After absorption with adult tissues, this antiserum specifically recognized antigens located in the cytoplasm of fetal pancreatic acini. All of the examined pancreatic tissues, ranging from 3 to 5 months of gestational age, showed a strong positive reaction of most of the acinar cells. The number of stained acini and the staining intensity gradually decreased from 5 months onwards and by the 7th–8th month only a few cells remained positive. Adult pancreas was completely negative as were a variety of normal adult and fetal tissues. This antiserum also reacted with tumor structures in 18/18 pancreatic adenocarcinomas as well as with pancreatic acini in the vicinity of tumor. Primary carcinomas of the liver, large bowel, stomach, breast, urinary bladder, lung and other localizations did not react with this antiserum. In some cases of chronic pancreatitis (3/12) a reaction was observed in a few acinar cells. Immunoblot assay after polyacrylamide electrophoresis revealed, in both fetuses and tumors, two main antigens of approximately 60 kDa and 110 kDa relative molecular weight. Several minor components were also observed.
Risk of colorectal cancer in adenoma‐bearing individuals within a defined population
Authors: Eide Tor J.
Published: February 2007

Assuming that all colorectal cancers develop from preexisting adenomas, the annual conversion rate, defined as the number of cancers occurring each year as a percentage of all adenoma‐bearing individuals, was determined. The number of adenoma‐bearing individuals in the living population of northern Norway was estimated in each cohort for the period 1974–76 by determining the prevalence of colorectal adenomas in an autopsy population of 271 consecutive cases, representative of the population of the area with regard to underlying causes of death. During the 10‐year period 1974–1983 a total of 656 colorectal cancers were recorded among an estimated number of 26,419 adenoma‐bearing individuals aged over 35 years. The annual conversion rate was found to be 0.25%, indicating that an average adenoma‐bearing individual is only at a moderate risk of colorectal cancer. The annual conversion rates for individuals having large adenomas, or adenomas with villous structures, or severe dyspla‐ sia were roughly estimated to be 3%, 17% and 37% respectively, assuming that colorectal cancer develops from one of these sub‐groups of adenomas only.
Evolution in the structure and distribution of 4F2‐antigen from the oncofetal to the adult phenotype of human fibroblasts
Authors: Azzarone B. , Eid P. , Malpiece Y. , Andeol Y. , Fauci A. S. , Suarez H.
Published: February 2007

The monoclonal antibody (MAb) 4F2 defines an oncofetal antigen in human fibroblastic cells. Two‐dimensional electrophoretic analysis reveals that tumor cell lines from mesenchymal tissues co‐express two or more heavy‐chain molecular variants of the antigen whereas the light subunit (41 kDa) is not affected. Among normal cells, only embryonic and newborn fibroblasts (from donors up to 20 days after birth) clearly co‐express two distinct molecular forms of the heavy chain with MW of 85 and 75 kDa, respectively. Cells derived from 3‐month‐old donors express detectable amounts of the 85 kDA but only faint traces of the 75 kDa subunit, while fibroblastic cells derived from donors older than 3 months seem to express only the 85 kDa subunit. Immunofluorescence analysis performed on adherent living cells shows that, in the first months after birth, there is a gradual evolution from the oncofetal to the adult phenotype also in the cell distribution of the 4F2. This evolution is reflected by a progressive disappearance of the 4F2 antigen from the cell membrane becoming, in adult normal cells, inaccessible to anti‐4F2 MAb. The existence of different molecular forms and different membrane positions of the 4F2 antigen could facilitate surveillance of morphological and structural changes in the evolution of human fibroblastic cells during the developmental process and neoplastic transformation.
Case‐control study on colorectal cancer and diet in marseilles
Authors: Macquart‐Moulin Geneviève , Riboli Elio , Cornée Jacqueline , Charnay Bernadette , Berthezene Patrice , Day Nicholas
Published: February 2007

A case‐control study of cancer of the colon and rectum was conducted in the Marseilles region of southern France. Cases (399) and a corresponding number of controls, matched for age and sex, were included, the controls being selected from patients undergoing functional re‐education for injuries or trauma which reduced their mobility. A dietary history questionnaire was used to determine the usual eating habits during the year preceding diagnosis for cases, or preceding interview for controls. The cases reported lower consumption of vegetables and oil than controls, but no differences were seen in the consumption of meat, bread, eggs or butter. The intake of several nutrients, particularly vitamins B2, B6, C, potassium, iron, magnesium and vegetable fibre, was lower among cases. However, when all these nutrients were analysed jointly and adjusted one for the other, only potassium retained a significant effect. This may be due to the high degree of colinearity between the estimated intake of many nutrients. No association was seen with fat or fibres from cereals.
A comparison of the in vivo uptake and metabolism of 3H‐oestrone and 3H‐oestradiol by normal breast and breast tumour tissues in post‐menopausal women
Authors: McNeill J. M. , Reed M. J. , Beranek P. A. , Bonney R. C. , Ghilchik M. W. , Robinson D. J. , James V. H. T.
Published: February 2007

After infusion of 3H‐oestradiol or 3H‐oestrone into post‐menopausal women with breast cancer, there was a significant uptake of both steroids by breast tumour and normal tissue. The proportion of oestrogen present in tumour tissue as 3H‐oestradiol after infusion of 3H‐oestradiol (89.4 ± 3.5%, mean ± SD, n = 4) was significantly higher (p < 0.001) than in normal breast tissue (72.8 ± 3.3%) obtained from the same women. Similarly, after infusion of 3H‐oestrone, the proportion of oestrogen present as 3H‐oestradiol (48.4 ± 14.4%) was significantly higher than in normal breast tissue (19.1 ± 6.4%). These results suggest that conversion of oestrone to oestradiol is enhanced in breast tumour tissue with little metabolism of oestradiol. This would account for the higher concentrations of oestradiol reported in breast tumour tissue in the presence of increased oestradiol 17β‐hydroxysteroid dehydrogenase activity.
Combined effect of ACNU and 5‐FU on rat glioma cells in spheroids and monolayer cultures
Authors: Kitahara Masakazu , Katakura Ryuichi , Mori Teruaki , Suzuki Jiro , Sasaki Takehito
Published: February 2007

Combined effects of ACNU and 5‐Fu on rat glioma clone‐6 cells grown exponentially as monolayers and in multicell spheroids (500–600 μm in diameter) were analyzed by the colony‐forming assay. Cells in spheroids with central necroses were more sensitive to ACNU than cells in monolayer, although a large fraction of the cells in these spheroids was resistant to 5‐Fu. The effect of ACNU was enhanced by the combined treatment with 5‐Fu for cells in both spheroids and monolayers, and was more remarkable when 5‐Fu was administered 24 hr prior to ACNU treatment. The enhancement was mainly due to a decrease in the number of spheroid‐cells resistant to 5‐Fu. Isobolo‐gram analysis indicated that the enhancement was supra‐additive when a low concentration of 5‐Fu was combined with a high concentration of ACNU. It is suggested that a low dose of 5‐Fu combined with a high dose of ACNU may preferentially kill more cells in solid tumors.
Anti‐tumor effects of interferon in mice injected with interferon‐sensitive and interferon‐resistant friend leukemia cells. IV. Definition of optimal treatment regimens
Authors: Gresser Ion , Belardelli Filippo , Maury Chantal , Tovey Michael G. , Maunoury Marie‐Thérèse
Published: February 2007

Mouse interferon α/β exerted a similar anti‐tumor effect in DBA/2 mice injected i.p. with Friend eryth‐roleukemia cells (FLC) either sensitive or resistant to interferon as determined by bothin vitroandin vivoassays. Using this tumor system we attempted to define optimal treatment regimens for interferon administration. Interferon was most effective when injected at the site of tumor inoculation rather than at a distant site. Two factors seemed of especial importance: the amount of interferon injected and the frequency of interferon administration. Thus, for daily administration of interferon, the antitumor effect was directly related to the amount of interferon injected. For a given total dose of interferon, repeated administration of small doses of interferon was more effective than administration of a larger dose at more widely spaced intervals. The anti‐tumor efficacy of interferon was independent of the number of FLC inoculated when 102to 105FLC were injected, but interferon treatment was less effective when 106or 107FLC were injected. The relevance of these results to the use of interferon in patients with cancer is discussed.
A verion‐like particle (VLP) released from an epstein‐barr virus‐producing cell sub‐line of P3HR‐1: Identification of a specific cellular DNA contained within VLPs
Authors: Yano Shoki , Miyoshi Isao , Ohtsuki Yuji , Koizumi Shigeki , Osato Toyoro
Published: February 2007

A vision‐like particle (VLP) was isolated from the culture fluid of P3HR‐1 cells. VLPs were separable from the enveloped Epstein‐Barr virus (EBV) through rate‐velocity sedimentation by virtue of their greater density. VLPs comprised electron‐dense nucleotide material and the surrounding round or oval envelope which ranged from 100 to 150 nm in diameter. The polypeptide constitution of VLPs was very similar to that of enveloped EBV by SDS‐polyacrylamide gel electrophoresis. VLPs contained reasonably specific cellular DNA (VLP DNA) with a sedimentation coefficient of 48S, which was cleaved into 5, 8, and at least 30 specific DNA fragments by digestion with BamHI, Sal, and EcoRI, respectively. Blot hybridization data suggested a limited heterogeneous genomic organization of VLP DNA and the presence of a VLP DNA‐related molecule(s) in human cord lymphocytes.