bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of the epstein‐barr virus (EBV)‐encoded membrane protein LMP1 impairs the In vitro growth, clonability and tumorigenicity of an EBV‐negative burkitt lymphoma line
Authors: Cuomo Laura , Ramquist Torbjörn , Trivedi Pankaj , Wang Fred , Klein George , Masucci Maria G.
Published: February 2007

In a previous study on several independently established Epstein‐Barr virus (EBV)‐converted sublines of the EBV‐negative Burkitt lymphoma (BL) line BL41, we found that expression of the virally encoded membrane protein LMP I was accompanied by reduced agarose clonability and tumorigenicity. In order to investigate whether LMPI can induce these phenotypic changes by itself, we have now studied the growth in suspension culture, the clonability in agarose and the tumorigenicity in immunosuppressed and SCID mice of 4 LMPItransfected sublines of BL4I that carry the gene under the control of the ZnSO4‐inducible metallothionein promoter. Expression of LMPI at levels comparable to those detected in EBV‐transformed lymphoblastoid cell lines (LCL) correlated with impairment of growth in suspension and reduction of clonability and tumorigenicity. Only minor changes were observed in transfectants expressing low LMPI levels. Upregulation of LMPI by ZnSO4 treatment of the low LMPI clone MTLM5 was accompanied by a slowing down of proliferation, increased cell clumping and decreased clonability. The results suggest that expression of LMP I at levels which are compatible with immortalization of normal B‐cells antagonizes the ability of BL cells to grow in vitro and in vivo, and illustrate a possible mechanism by which down‐regulation of this viral antigen may favor tumorigenicity in EBV‐carrying BLs. © 1992 Wiley‐Liss, Inc.
MHC class‐I‐restricted auto‐tumor‐specific CD4+CD8 −T‐cell clones established from autologous mixed lymphocyte‐tumor‐cell culture (MLTC)
Authors: Wang P. , Vànky F. , Klein E.
Published: February 2007

Autologous mixed lymphocyte‐tumor cell cultures (MLTC) were initiated with cytokine (IFNγ and TNFα)‐treated ex‐vivo tumor cells of lung, ovarian, breast and stomach carcinomas. The cytokine‐treated tumors expressed class‐l but not class‐M molecules. Although the proportion of CD8+ lymphocytes increased in the bulk culture of MLTCs, in 5/7 experiments the majority of the established T‐cell clones were CD4+. Among the CD8+ clones a high proportion (77%) was cytotoxic, while the proliferative response was more frequent among the CD4+clones (70%). In 4/26 cytotoxic T‐lymphocyte (CTL) clones (3/17 CD4+ and I /9 CD8+), derived from a patient with class l+class II− stomach carcinoma, lysis was restricted to the autologous tumor cells. These auto‐tumor‐specific clones did not lyse the autologous ConA blasts, the 5 allogeneic ex‐vivo tumors, the NK‐sensitive K562 or the relatively sensitive Daudi cells. The cytotoxicity of these clones was inhibited by pre‐incubation of the tumor cells with W6/32 (α‐class I) MAb, or by preincubation of the lymphocytes with OKT3 (α‐CD3) MAb. The α‐CD4 (OKT4) MAb had only a marginal effect on the CD4+clones, while the lytic function of the CD8+ clone was inhibited by the α‐CD8 (OKT8) MAb. The 3 CD4+CTL clones also responded with proliferation to the autologous tumor cells. This proliferative response was inhibited by the presence of W6/32 MAb. Our results indicate that the auto‐tumor lysis exerted by CD4+ CTL clones was restricted by the class‐l antigens, and that the CD4 molecules of the clones were not essential for the lytic interaction. © 1992 Wiley‐Liss. Inc.
Growth in serum‐free medium of NIH3T3 cells transformed by the EJ‐H‐ras oncogene: Evidence for multiple autocrine growth factors
Authors: Pironin Martine , Clément Gilles , Benzakour Omar , Barritault Denis , Lawrence' David , Vigier Philippe
Published: February 2007

Rodent fibroblastic cells transformed by ras oncogenes can grow in serum‐free (S−) medium. We have studied clonal lines of mouse NIH3T3 fibroblasts transfected with the EJ‐H‐ras oncogene, and observed that practically all become independent of exogenous growth and attachment factors shortly after transfection. Moreover, all the clones tested soon form anchorage‐independent (Al) colonies in S− medium, and most give rise to spheroids able to grow in suspension. The cell‐conditioned S−medium of the transformed (TR) cells stimulates autocrinally the Al and anchored growth of these cells, in the absence of serum, and it contains growth factors related to TGF‐α (or EGF), PDGF and bFGF, and other uncharacterized factors. Some of these factors are not found, or are found only in very small amounts, in the S− medium of non‐transformed NIH3T3 cells, which also stimulates the growth of the TR cells, in the absence of serum. In addition, the TR cells contain 4‐6 times more cell‐associated bFGF than the non‐transformed cells and release more latent TGF‐β activatable by acid treatments. However, no active TGF‐β is secreted by either cell type. Activated TGF‐β and pure TGF‐β I stimulate the growth of the anchored TR and NIH3T3 cells, but inhibit the Al growth of the TR cells. Another inhibitor of this growth is also found in the concentrated medium of the NIH3T3 cells. © 1992 Wiley‐Liss, Inc.
Hyaluronan (hyaluronic acid) and hyaluronectin in the extracellular matrix of human breast carcinomas: Comparison between invasive and non‐invasive areas
Authors: Bertrand P. , Girard N. , Delpech B. , Duval C. , D'Anjou J. , Dauce J. P.
Published: February 2007

We performed quantitative determination of the distribution of hyaluronan (hyaluronic acid, HA) and the HA‐binding protein, hyaluronectin (HN), 2 components of the extracellular matrix of tumor desmoplasia, within 71 human breast carcinomas. Results showed that HA and HN were more elevated in tumoral than in non‐tumoral adjacent tissue, and that the peripheral invasive area of tumors contained increased levels of HA and HN as compared with the central non‐invasive area (p < 10–3 and p < 10–5 respectively). HN and HA levels of 61 ductal carcinomas were related to the histological grade of tumors; no significant difference was found between grades for HA; HN was found to be significantly lower in grade III than in grade II tumors (p < 0.01). HA and HN rates were correlated in grade I and grade II tumors and were not correlated in grade III. Mean percentage of HA saturation level by HN for whole tumors was found to be less than 4%, indicating that HA is essentially free of proteins and could be used as a target for cancer diagnosis or therapy. © 1992 Wiley‐Liss, Inc.
Breast cancer in a low‐risk population. The influence of age and menstrual status on disease pattern and survival in Saudi Arabia
Authors: Al‐Idrissi Hassan Y. , Ibrahim Ezzelin M. , Kurashi Nabil Y. , Sowayan Saleh A.
Published: February 2007

Data on 130 women with invasive breast cancer, seen at our institution between April 1981 and November 1990, were retrospectively reviewed to assess the influence of age and menstrual status on the pattern and prognosis of their disease. Patients were mostly young (median age 40 years) and in 21 patients (16%) the diagnosis was established at the age of 30 years or younger. Eighty‐six patients (66%) were under 50 and 82 patients (63%) were pre‐menopausal. Pre‐menopausal patients were more numerous than expected in stage III and less numerous in stage II. On the other hand, the differences between observed and expected values for both stage I and stage IV in the 2 menopausal groups were not significant. Comparedwithpost‐menopausal patients, pre‐menopausal patients with 1–3 or > 3 positive lymph nodes were more numerous than expected. Differences between pre‐menopausal and post‐menopausal patients have persisted after categorizing patients into 2 age‐groups with a cut‐off point at 50 years. Comparable initial assessment and therapeutic modalities were offered to the 2 menopausal groups. At the time of analysis (January, 1991) all patients had a complete follow‐up. Over a median follow‐up of 46 months, the overall median survival (±SE) was 85.7 (±4.4) months with a survival probability (±SE) at 5 years of 62% (±5%). The proportional hazard model of Cox has identified advanced stage (stages III and IV) and involvement of lymph nodes as the only independent adverse predictors of survival with estimated hazard rates of 2.9 and 2.8, respectively. Unadjusted analysis, adjusted analysis and stratified survival functions failed to reveal any survival difference based on age or menstrual status. We conclude that, in a low‐risk population and despite apparent baseline differences in demographic and disease characteristics between pre‐menopausal and post‐menopausal breast cancer patients, neither age nor menopausal status had a significant influence on survival. Our results should guide future cancer‐care programs in Saudi Arabia. © 1992 Wiley‐Liss, Inc.
The role of mucin in colon‐cancer metastasis
Authors: Schwartz Bertha , Bresalier Robert S. , Kim Young S.
Published: February 2007

Mucinous colorectal cancer often presents at an advanced stage. We have previously observed that mucin production by human colon‐cancer cells correlates with their ability to colonize the liver in experimental animal models. The present study was undeitaken in order to further elucidate the mechanisms by which production of mucin by colon‐cancer cells affects metastasis. Cell lines showing high mucin production (HMP) (HM 7, HM 3 and LS LiM 6) demonstrated increased adherence to basement membrane proteins and invaded a reconstituted basement membrane to a greater extent than their counterpart cell lines showing low mucin production (LMP) (LS174T and LM 12). Adherence of the LMP parental cell line LS 174T to various matrix proteins was potentiated by the addition of purified human colon‐cancer mucin in a dose‐dependent fashion. HMP cell lines secreted more proteolytically active type‐IV collagenase than LMP lines, and collagenase activity was further stimulated by purified mucin in a dose‐dependent manner. Specific inhibition of mucin O‐glycosylation by benzyl‐α‐N‐ acetylgalactosamine significantly affected each of the metastasis‐ related events, with the greatest effect on the HMP cell lines. The present data further indicate that cumin may play an important role in the metastutic process. © 1992 Wiley‐Liss, Inc.
Morphological, immunohistochemical and biochemical characterization of 6 newly established human ovarian carcinoma cell lines
Authors: Möbus V. , Gerharz C. D. , Press U. , Moll R. , Beck T. , Mellin W. , Pollow K. , Knapstein P. O. , Kreienberg R.
Published: February 2007

Six permanent human tumor cell lines (OV‐MZ‐1 to 6) were established from 6 patients with serous adenocarcinomas of the ovary. These cell lines were derived from both solid tumors and ascites, from pre‐treated and untreated patients, and are available over a range of in vitro passage numbers. The tumor cells grow its monolayers and develop foci of “piled‐up” cells in confluent cultures. Flow cytophotometry showed that all the lines exhibited DNA hyperdiploidy with DNA tetraploidy in one cell line and DNA aneuploidy in the other cell lines. The mean population doubling time ranged from 24 to 52 hr. Transmission electron microscopy demonstrated that the tumor cells of all cell lines exhibited features of epithelial differentiation such as desmosomes and intracellular gland‐like lumina. Immunocy‐ tochemical analysis showed that the co‐expression of cytokeratins and vimentin, which is a feature of ovarian serous cystadeno‐ carcinomas; in situ, was fully preserved in the majority of cell lines. The main cytokeratin polypeptides expressed were numbers 7, 8, 17, 18 and 19. The tumor‐associated antigen CA‐125, but not CEA, was shed in the culture supernatant. This was in accordance with FACScan analysis of the cell lines and the level of CA‐125 and CEA in the patients' serum. The estrogen and progesterone receptors were negative both in the cell lines and in the original tumors. These new ovarian carcinoma cell lines will be valuable models for further investigations into a variety of biological properties. © 1992 Wiley‐Liss, Inc.
Establishment and characterization of human uterine leiomyosarcoma heterotransplanted into nude mice
Authors: Sawada Masumi , Terada Nobuyuki , Yamamoto Reiko , Nishizawa Yasuko , Wada Akira , Mori Yoichi , Sakamoto Hiromi , Tanizawa Osamu
Published: February 2007

Human uterine leiomyosarcoma is a rare gynecological malignancy with a generally poor prognosis. We have established a human uterine leiomyosarcoma tumor line in nude mice, designated UTS‐1, and describe the characteristics of this tumor. The UTS‐1 tumor doubled in 12.1 days and retained the histological characteristics of leiomyosarcoma, even after 14 serial generations. Ultrastructurally, the tumor is characterized by nuclear pleomorphism typical of smooth muscle, intracyto‐plasmic filaments with dense bodies, a relative paucity of micropinocvtotic vesicles, and an incomplete external lamina. Immunohistochemically, the UTS‐1 cells reacted with antibodies against vimentin, desmin, smooth‐muscle actin and myosin, but not with antibodies against keratin, CEA and S‐100 protein. Serum levels of AFP, CAI25, CEA and SCC ranged within normal limits in tumor‐bearing mice. The serum level of immu‐nosuppressive acidic protein correlated well with an activity of the tumor. Estrogen and progesterone receptors were not detected in the tumor. Chromosomal analysis showed a human karyotype with some marker chromosomes and a modal number of 85 chromosomes. The UTS‐1 tumor should prove a useful model to explore the biological characteristics and treatment of human uterine leiomyosarcoma. © 1992 Wiley‐Liss, Inc.
Protein‐kinase‐C inhibitor calphostin C reduces B16 amelanotic melanoma cell adhesion to endothelium and lung colonization
Authors: Liu Bin , Renaud Colette , Nelson Kevin K. , Chen Yong Q. , Bazaz Rajesh , Kowynia Julie , Timar Jozsef , Diglio Clement A. , Honn Kenneth V.
Published: February 2007

We recently reported that the Ca2+‐and phospholipids‐dependent protein kinase, protein kinase C (PKC), was involved in rat Walker carcinosarcoma cell adhesion to large‐vessel endothelium. We extended our studies to explore the role of this kinase in the adhesion to small‐vessel endothelium and lung colonization of murine B16 amelanotic melanoma (B16a). Sub‐populations of B16a cells, which differ in lung‐colonization potentials, were isolated by centrifugal elutriation from solid tumors. In this study, we demonstrate that cells from a high metastatic sub‐population (HM340), when compared with cells from a low metastatic sub‐population (LM180), exhibit elevated levels of total cellular as well as membrane‐bound PKC. The increase in PKC in cells from the HM340 correlates positively to their increased ability to adhere to murine pulmonary‐microvessel endothelial‐cell monolayer, and to form pulmonary colonies in syngeneic mice. Calphostin C, a potent and selective PKC inhibitor, decreases in a dose‐dependent manner the adhesion to endothelium and the lung colonization of cells from both the low and the high metastatic sub‐populations with IC50 at sub‐micromolar concentrations. In conclusion, our results suggest that PKC may be a key element in regulating tumor‐cell metastasis and that PKC inhibitors may be anti‐metastatic agents. © 1992 Wiley‐Liss, Inc.
Receptors for hormones and growth factors and (ONCO)‐gene amplification in human ovarian cancer
Authors: Berns Els M. J. J. , Klijn Jan G. M. , Henzen‐Logmans Sonja C. , Rodenburg Cees J. , van der Burg Maria E. L. , Foekens John A.
Published: February 2007

Receptor status and gene amplification were studied in advanced human ovarian adenocarcinoma tissues, borderline and benign ovarian tumours and normal ovarian tissues. Sixty‐five percent (53/82) of ovarian adenocarcinomas, 57% (8/14) of benign/borderline tumours and only 31% (5/16) of normal ovarian tissues studied showed specific 125I‐EGF (epidermal growth factor) binding (median: 17; 10; and 0 fmol EGF‐R/mg protein, respectively) and a significant increase in progesterone receptor (PgR) levels was observed in these groups (median: 5; 33; and 152 fmol/mg protein, respectively). No correlations were observed between the levels of EGF‐R and the levels of either oestrogen receptors (ER) or PgR. All membrane samples of 25 adenocarcinomas studied by Scatchard analysis were positive for insulin‐like growth‐factor‐1 receptors (IGF‐1 ‐R) and contained higher IGF‐I‐R levels than membranes of 10 normal ovarian tissues, of which 9 were positive (median: 64 and 26 fmol IGF‐I‐R/mg membrane protein, respectively). Also, as measured by autoradiography, 37 adenocarcinoma tissues showed a higher expression of IGF‐I‐R (1.5+ to 4+) than sections derived from 10 normal ovarian tissues (I +). 125I‐IGF‐I binding was predominantly associated with epithelial tumour cells, the surrounding connective tissue was negative and in several samples high expression of IGF‐I‐R was found in areas of tumour necrosis. Southern blot analysis of DNAs isolated from 25 ovarian adenocarcinomas revealed no amplification of the IGF‐I ‐R or the EGF‐R gene. The HER2/neu gene was amplified only in 2 out of 3 histologically confirmed endometrioid adenocarcinomas studied but not in 22 other tumours. An amplification of the c‐myc gene was observed in 28% (7/25) of the tumours. All c‐myc‐amplified tumours were PgR‐negative. No rearrangement was observed for any of the genes studied. In conclusion, ovarian adenocarcinoma tissues show a decrease in PgR levels and an increased expression of IGF‐I‐R and EGF‐R, in the absence of gene amplification, when compared to benign/borderline ovarian tumours and normal ovarian tissues. In addition, amplification of the c‐myc and HER2/neu genes, without rearrangement of these genes, occurs in a minority of these tumours. © 1992 Wiley‐Liss, Inc.
Influence of recombinant human interleukin (IL)‐7 on disease progression in mice infected with friend virus complex
Authors: Lu Li , Zhou Zhen , Wu Bo , Xiao Mang , Shen Rong‐Nian , Williams Douglas E. , Kim Young‐Junc , Kwon Byoung S. , Ruscetti Sandra , Broxmeyer Hal E.
Published: February 2007

Recombinant human (rhu) IL‐7 was evaluated for its influence on disease progression in mice infected with the polycythemia‐inducing strain of the Friend virus complex (FVC). DBA/2 mice were injected i.v. with FVC, and then treated s.c. with rhuIL‐7. IL‐7 significantly prolonged survival time and decreased spleen focus‐forming virus (SFFV) levels, expression of SFFV mRNA and SFFV protein production in FVC‐infected mice. IL‐7 did not appear to directly inactivate SFFV. Although both splenic weight and cellularity in FVC‐infected mice treated with IL‐7 were higher than those of normal mice, they were respectively 58% and 66% lower than those of the untreated FVC‐infected mice. NK‐cell activity was substantially lower in FVC‐infected mice than in normal mice, while IL‐7 restored NK‐cell activity to normal levels. IL‐6 and IFN‐γ levels were markedly reduced in FVC‐infected mice compared to normal mice, but treatment of FVC‐infected mice with IL‐7 restored these cytokine levels. While the actual mechanisms of these effects are not yet known, the results suggest the potential therapeutic efficacy of IL‐7 for certain hematopoietic and viral disorders, possibly mediated through an action on accessory cells and cytokine production. © 1992 Wiley‐Liss, Inc.
Phenotype‐specific “tissue” transglutaminase regulation in human neuroblastoma cells in response to retinoic acid: Correlation with cell death by apoptosis
Authors: Piacentini Mauro , Annicchiarico‐Petruzzelli Margherita , Oliverio Serafina , Piredda Lucia , Biedler June L. , Melino Gennaro
Published: February 2007

Neuroblastomas in culture are characterized by the presence of 2 morphologically and biochemically distinct phenotypes (i.e., neural “N‐type” and flat substrate‐adherent “S‐type”) which undergo transdifferentiation. Human neuroblastoma SK‐N‐BE(2) cells differentiate toward a neural phenotype upon retinoic acid (RA) treatment. However, we recently showed that, during the RA treatment, a subset of SK‐N‐BE(2) cells undergo apoptosis; these cells specifically express a high “tissue” transglutaminase (tTG) level. This study was undertaken to investigate the cellular and molecular basis of the action of retinoic acid on apoptosis in human neuroblastoma cells. As a biochemical marker of the phenomenon we studied the tTG gene expression in the parental line SK‐N‐BE(2) and in 2 clones which stably express neuroblastic [BE(2)‐M17] and substrate‐adherent [BE(2)‐C] features, respectively. Data showed a differential phenotype‐specific regulation of tTG gene expression. In fact, RA treatment enhanced tTG expression and apoptotic index in the flat substrate‐adherent variant, whereas, in cells expressing the neural phenotype, very low tTG expression and apoptosis were found. Northern‐blotting analysis revealed that the substrate‐adherent cells had a basal 3‐fold higher level of tTG mRNA. An increase in tTG mRNA major transcript levels (3.7 kb) occurred within a few hours of exposure to RA in both the phenotypic variants. By contrast, tTG protein level was very low in the cell expressing the neuronal phenotype, even after prolonged exposure to RA. Immunohistochemical analysis indicated that tTG protein, in addition to mature apoptotic cells, was specifically localized in the flat substrate‐adherent variant both in the wild‐type and in the BE(2)‐C clone. These findings suggest that the ability to undergo apoptosis in the neuroblastoma cells is associated with the expression of a non‐neuronal neuroectodermal (substrate‐adherent cells) immature phenotype. © 1992 Wiley‐Liss, Inc.
Influence of glucose on metabolism and growth of rat glioma cells (C6) in multicellular spheroid culture
Authors: Acker H. , Holtermann G. , Bölling B. , Carlsson J.
Published: February 2007

The metabolism and growth of rat glioma C6 cells in multicellular spheroid culture depended strongly on the glucose supply. A low glucose level (0.1 g/l) in the culture medium reduced lactate production, increased oxygen consumption and diminished hydrogen ion production under normoxia as well as hypoxia. A high glucose level (10 g/l glucose) increased lactate production, had no significant influence on oxygen consumption and increased the hydrogen ion production under hypoxia. Hydrogen ion release from cells under normoxic and hypoxic conditions could be significantly diminished by amiloride (1 mM), indicating the involvement of the Na+/H+ exchanger. The growth of the C6 spheroids was enhanced under low glucose conditions, possibly due to the more physiological extracellular pH in the deeper regions of the spheroids. The growth was inhibited under high glucose conditions, which seemed to be toxic due to a massive hydrogen production giving acidosis. The glucose supply strongly influenced the local hydrogen ion production inside the C6 spheroids and this might in turn lead to changes in the response to different therapeutic modalities. © 1992 Wiley‐Liss, Inc.
Transforming activity and the level of tax protein: Effect of one point mutation in HTLV‐I tax gene
Authors: Sakurai Mamoru , Yamaoka Shoji , Nosaka Tetsuya , Akayama Masayuki , Tanaka Atsushi , Maki Masatoshi , Hatanaka Masakazu
Published: February 2007

The transcriptional trans‐activator molecule of HTLV‐I, Tax, is known to transform rodent fibroblasts. A revertant clone expressing Tax was obtained by treating transformed Rat‐1 cells harboring a single copy of the tax gene with a mutagen. Sequence analysis of the tax gene of the revertant clone revealed that it had one point mutation at codon 12(CTT × TTT), resulting in a change from Leu to Phe. The colony‐forming efficiencies of the cells transfected with the mutant tax gene (mu71tax) were significantly lower than those transfected with the wild‐type by the soft‐agar method. This difference was shown to be due to the instability of mu71Tax. © 1992 Wiley‐Liss, Inc.
The H‐ras oncogene regulates expression of 70‐ and 45‐kDa cell‐surface molecules whose expression correlates with tumor‐cell immunogenicity
Authors: Gopas Jacob , Ehrlich Tali , Cohen Orit , Wishniak Orna , Tainsky Michael A. , Segal Shraga , Rager‐Zisman Bracha , Isakov N.
Published: February 2007

The effects of the H‐ras oncogene on fibroblast cell tumorigenicity and immunogenicity was studied in transfectants of the BALB/c 3T3 clone A31 fibroblastoid cell‐line. Cells that were transfected with MC29‐LTR‐H‐ras (98/6) or MC29‐LTR‐v‐myc + H‐ras (98/4v) and were inoculated into syngeneic BALB/c mice were tumorigenic in 100% and 60% of animals respectively. By contrast, transfectants containing the pSV2neo plasmid alone (98/1) displayed normal characteristics both in vitro and in vivo. Inoculation of mice with mitomycin‐C‐treated 98/1 or 98/4v cells induced an effective protective immunity to a challenge of live 98/4v cells, and a partial immunity against 98/6 cells. Mitomycin‐C‐treated 98/6 cells failed to render immunity against a challenge of either 98/6 or 98/4v cells. To correlate immunogenicity and tumorigenicity of the different cell types with cell‐surface‐antigen expression, we prepared MAbs against 98/4v cells in syngeneic mice. Immunohistochemical and immunoblot analysis revealed that MAbs 102 and 104 recognized 2 protein bands of 70 and 45 kDa respectively, which were expressed predominantly in 98/1 and 98/4v cells. A third immunoreactive protein band of 44 kDa that reacted with MAb 6 was expressed at a similar cell‐surface density on all cell types. Cell‐differentiation‐inducing agents, such as DMSO, retinoic acid or sodium butyrate, were all found to induce 98/6 cell flattening and morphological changes toward a normal phenotype that were followed by up‐regulation of the 70‐ and 45‐kDa antigens. The results suggest that regulation of expression of the 70‐ and 45‐kDa molecules is affected by H‐ras, and that expression of these cell‐surface molecules may be relevant to tumor cell immunogenicity. © 1992 Wiley‐Liss, Inc.
Ornithine decarboxylase assay in human colorectal mucosa. Methodologic issues of importance to quality control
Authors: Garewal Harinder S. , Sloan Donna , Sampliner Richard E. , Fennerty Brian
Published: February 2007

Ornithine decarboxylase may be a useful biomarker for risk of neoplasia in colorectal tissues. Investigators have reported enzyme activities varying by as much as 10‐ to 20‐fold using variations of the usual 14CO2 release assay. We have examined the effect of different methodologic factors on calculated ornithine decarboxylase activity. Major effects on the assay result (>20% change) were produced by: (I) use of Tris vs. phosphate buffer, the former yielding 1.5‐ to 4‐fold greater activity; (2) protein content of the reaction mixture with significant error if <50 μg; (3) use of α‐difluoro‐methylornithine‐inhibited blank versus buffer‐only blank. Other changes in assay conditions, including addition of sucrose, detergent, protease inhibitors, specific activity of 14C‐ornithine, the nature of the trapping agent used, and incorporation of a sonication step, did not have a significant effect on ODC quantification (≤20%). Thus, seemingly minor variations in assay conditions can greatly affect the results, which may provide a partial explanation for the variability of ODC activities reported in the literature. Strict quality control measures are mandatory in the interpretation of clinical observations utilizing this marker as an endpoint. © 1992 Wiley‐Liss, Inc.
Secretion of gelatinases and tissue inhibitors of metalloproteinases by human lung cancer cell lines and revertant cell lines: Not an invariant correlation with metastasis
Authors: Zucker Stanley , Lysik Rita M. , Malik Masood , Bauer Brett A. , Caamano J. , Klein‐Szanto Andres J. P.
Published: February 2007

Numerous studies have reported a correlation between production of 72‐kDa (MMP‐2) and 92‐kDa (MMP‐9) type‐IV collagenases/gelatinases and the metastatic potential of cancer cells. An abrogating effect of tissue inhibitors of metalloproteinases (TIMP‐1 and TIMP‐2) on metastases has also been noted. In this report we have used sensitive enzyme‐linked immunoas‐says to measure MMP‐2, MMP‐9, TIMP‐1 and TIMP‐2 levels in eight human lung‐cancer cell lines which were characterized for biological behavior in nude mice. We demonstrated that the Calu‐6 and A549 cell lines with the highest metastatic, invasive and tumorigenic potential secreted the highest levels of MMP‐2. MMP‐9 and TIMP‐1 secretions were comparatively low in all cell lines. TIMP‐2 secretion, which exceeded MMP‐2 secretion for all cell lines, did not correlate with metastatic potential. To further explore these correlations, the metastatic Calu‐6 cell line was transfected with a K‐rev‐I cDNA expression construct. The K‐rev revertant cell lines demonstrated a more differentiated phenotype and were less tumorigenic, invasive and metastatic in nude mice. Nonetheless, the Calu‐6 revertant cell lines secreted higher levels of MMP‐2 than the parent cell line. In conclusion, invasion and metastasis by lung‐cancer cells requires not only enhanced MMP production, but also other less well‐understood tumorigenic characteristics. The multiplicity of factors required by cancer cells for dissemination helps to explain the minute fraction of cancer cells from a primary tumor that ever develop into a metastasis. © 1992 Wiley‐Liss, Inc.
A basement‐membrane permeability assay which correlates with the metastatic potential of tumour cells
Authors: Parish Christopher R. , Jakobsen Karen B. , Coombe Deirdre R.
Published: February 2007

We describe an in vitro assay for measuring the ability of tumour cells to permeabilize basement membranes, using transwell chambers coated with the reconstituted basement membrane, matrigel. Unlike previous matrigel‐based procedures which quantified passage of tumour cells across a matrigel barrier, the new assay measures the ability of tumour cells to degrade the basement membrane and increase the diffusion rate of fluorescent (FL) dextran through the barrier. The procedure has the major advantage that permeability can be rapidly and accurately quantified, either by fluorometry or by the use of radiolabelled dextran, thus avoiding tedious and subjective scoring methods. Optimal conditions for the assay are described. In addition, it is demonstrated that the assay can clearly discriminate between metastatic and non‐metastatic tumour cell lines, metastatic tumours permeabilizing the basement membrane and non‐metastatic counterparts failing to do so. A range of enzyme inhibitors suggested that the increase in basement‐membrane permeability caused by the metastatic mammary adenocarcinoma 13762 MAT is probably dependent upon the synergistic action of several degradative enzymes, namely proteases, type‐IV collagenase, and heparanase. Furthermore, the ability to permeabilize the basement membrane was dependent upon intact tumour cells; tumour cell extracts, lysates and supernatants were inactive. © 1992 Wiley‐Liss, Inc.
Isolation of two keratinocyte cell lines derived from HPV‐positive dysplastic vaginal lesions
Authors: Hietanen Sakari , Auvinen Eeva , Grénman Seija , Lakkala Taina , Sajantila Antti , Klemi Pekka , Mäenpää Juhani
Published: February 2007

Explant cultures were started from human papillomavirus (HPV)‐infected genital lesions in order to isolate and propagate abnormally differentiating cells from squamous intraepithelial neoplasia. A medium with high calcium concentration was used to induce terminal differentiation of cells from surrounding normal epithelium. Two cell lines with extended life‐spans were established. The UT‐DEC‐I cell line was derived from an HPV‐33‐positive mild vaginal dysplasia (VAIN I). In cultured UT‐DEC‐I cells, HPV 33 DNA was detected with Southern‐blot hybridization and the polymerase chain reaction (PCR) technique. The restriction pattern of HPV 33 changed during early passages and flow cytometric analysis detected a decrease in chromosomal DNA content. HPV 33 RNA from the E6‐E7 region could be amplified by PCR at late passage. UT‐DEC‐2 cell line was derived from an HPV‐16‐positive moderate vaginal dysplasia (VAIN II). HPV 16 DNA was also detected in cultured cells by the PCR technique. The senescence of normal keratinocytes and growth selection in favor of aneuploid cells was observed by flow cytometric analysis at subsequent passages. Karyotype analysis showed clonal chromosomal abnormalities in both cell lines. To date, UT‐DEC‐1 cells have undergone 40 and UT‐DEC‐2 cells 25 passages. This study shows that the isolation of HPV‐infected dysplastic cells can be achieved by culturing the cells in a medium with high calcium concentration. The cell lines presented provide the opportunity of evaluating the early stages of squamous‐cell carcinogenesis. © 1992 Wiley‐Liss, Inc.
Insulin‐like growth factor II in two human colon‐carcinoma cell lines: Gene structure and expression, and protein secretion
Authors: Lambert Sylvie , Carlisi Agnes , Collette Julien , Franchimont Paul , Gol‐Winkler Rosita
Published: February 2007

Abnormal expression and structural modification of the IGF‐II gene in correlation with high IGF‐II production have recently been described in human colorectal tumors in comparison with normal adjacent tissues. Here, we have studied IGF‐II in 2 human colon‐carcinoma cell lines, HT29 and COLO 320DM. RFLP analyses revealed no apparent alteration at the IGF‐II locus in these 2 cell lines. HT29 cells weakly expressed IGF‐II mRNA in comparison with the high over‐expression previously observed in some colorectal tumors, and only the 4.8‐kb mRNA species was present. In addition, the serum‐free medium conditioned by HT29 cells contained high IGF‐II levels (approx. 900 ng/ 108 cells), as measured in a specific IGF‐II radioimmunoassay (RIA). After chromatography on Bio‐Gel P‐60, we observed that 64% of the total IGF‐II secreted by HT29 cells were present as a high‐molecular‐weight form of about 17 kDa. In contrast, no detectable expression of the IGF‐II gene was observed in COLO 320DM cells, and low IGF‐II levels were secreted by these cells in the serum‐free medium, with only 9% total IGF‐II represented by the large species. © 1992 Wiley‐Liss, Inc.
Experimental photodynamic therapy with a copper metal vapor laser in colorectal cancer
Authors: Lantz J‐M. , Meyer C. , Saussine C. , Leberquier C. , Heysel F. , Miehe J. , Marescaux J. , Sultan R. , Kedinger M.
Published: February 2007

In an attempt to define the best conditions for an adjunctive treatment of residual colonic microtumors by photodynamic therapy (PDT), an experimental model has been defined. S.c. HT29 colonic‐cancer‐cell tumors grown in nude mice were used and, 48 hr after i.p. administration of 30 mg/kg Photofrin (PH), laser illumination was performed with 75 or 150 Joules/cm2. The efficiencies of 2 lasers, the classically used rhodamine laser (RL) and a copper metal vapor laser (CMVL), were compared. The effects of PDT were assessed by histological and immunocytochemical (detection of a digestive enzyme, dipeptidyl‐peptidase IV, as a marker of cell viability) follow‐up and by the growth curve of the tumors after illumination. We conclude that, although the depth of necrosis resulting from PDT was nearly 3 mm at 75 J/cm2 and nearly 4–5 mm at 150 J/cm2 with both lasers, complete necrosis was obtained only with the CMVL at 150 J/cm2 (in 50% of the tumors). Under the other conditions, a layer of unaffected cells persisted at the pole opposite to laser illumination, resulting in growth curves lower than but parallel to those of the controls. Analysis of drug concentrations in the tumors and various organs, 48 hr after injection, i.e., at the time of laser illumination, revealed the presence of 21 μg/g dry weight PH in the tumors. The tumor vs. host‐organ ratios were equal to or higher than 1 for the small bowel, colon, stomach, lung, skin and muscle. In contrast, the ratios were below 1 for the spleen, pancreas, kidney and liver. © 1992 Wiley‐Liss, Inc.
Characterization of a lung‐cancer‐associated auto‐antigen
Authors: Zeng Carl Q. Y. , Alpert Lesley C. , Alpert Elliot
Published: February 2007

An antigen, protein X (Px), was purified from immune complexes isolated from malignant pleural effusions from patients with adenocarcinoma of the lung by EDTA treatment, PEG 8000 precipitation, protein A affinity chromatography, and Sephadex G‐200 separation in the presence of 3 M NaCl. The purified antigen had a Mr 17,000 by SDS‐PAGE, and consisted of isoelectric species of pl 6.3 and 6.6. Purified Px recombined with Ig isolated from pleural fluids from patients with lung adenocarcinoma, but not with Ig from patients with breast carcinoma. Using an autologous human and heterologous chicken antibody, Px was found, by immunohistology, in the cytoplasm of some of the well‐differentiated lung adenocarcinoma cells, but was not seen in normal lung or a variety of other malignant tissues. A liquid‐phase competitive‐inhibition RIA was developed. Over 30 ng/ml of Px were found in 9 of 15 pleural fluids from patients with lung carcinoma, none of 20 from patients with breast, ovary, stomach or colon cancer, and in 3 of 15 patients with unknown primary tumor. Our data suggest that Px may be a lung‐cancer‐associated autoantigen which can elicit a host humoral response in vivo. © 1992 Wiley‐Liss, Inc.
Carcinogenic effect of tobacco smoking and alcohol drinking on anatomic sites of the oral cavity and oropharynx
Authors: Boffetta Paolo , Mashberg Arthur , Winkelmann Regina , Garfinkel Lawrence
Published: February 2007

A series of 359 male patients with 424 cancer lesions of the oral cavity and oropharynx identified at a US Department of Veterans Affairs Medical Center were divided according to site of origin of the lesion and compared with 2,280 controls from the same hospital with respect to exposure to tobacco smoking and alcohol drinking. Sites of origin were: floor of the mouth (153), oral tongue (50), anterior tonsillar pillar (49), soft palate (44), lingual aspect of retromolar trigone (11), alveolar ridge (5), buccal mucosa (4), and hard palate (2). Forty‐one patients had cancers in multiple sites. Tobacco smoking was more strongly associated with soft‐palate lesions than with lesions in more anterior sites. Patients with cancer of floor of the mouth and oral tongue had higher odds ratios for alcohol drinking than subjects with cancers of other sites. This study supports the hypothesis of the carcinogenic effect of tobacco smoke and alcoholic beverages on the oral mucosa through direct contact. © 1992 Wiley‐Liss, Inc.
Expression of the matrix Gla protein in urogenital malignancies
Authors: Levedakou Eleni N. , Strohmeyer Torsten G. , Effert Peter J. , Liu Edison T.
Published: February 2007

Matrix Gla protein (MGP), is a vitamin‐K‐dependent protein which is synthesized in a variety of tissues such as lung, heart, kidney, cartilage and bone. The function of MGP in these tissues is unclear. We have previously reported elevated MGP mRNA levels in a breast‐cancer cell line, 600PEI, as compared to normal breast epithelium. Here we describe high MGP expression in primary renal‐cell carcinomas, prostate carcinomas and testicular germ‐cell tumors, as determined by Northern analysis. MGP was over‐expressed in 21 out of 28 patients with renal‐cell carcinoma, and in 16 out of 29 patients with testicular germ‐cell tumors, as compared to matched normal tissues. For the renal‐cell carcinomas, a statistically significant inverse correlation was observed between the level of MGP expression and tumor size, lymph‐node metastasis and tumor grade. MGP was also highly expressed in 13 primary prostatic carcinomas as compared to prostate cell lines derived from metastatic tumors, and to lymph‐node metastasis. Our findings indicate that the loss of MGP expression may be associated with tumor progression and metastasis. © 1992 Wiley‐Liss, Inc.
International variations in the incidence of neuroblastoma
Authors: Stiller C. A. , Parkin D. M.
Published: February 2007

The International Agency for Research on Cancer has coordinated a worldwide study of childhood cancer incidence, with data from over 50 countries. We present here the results for neuroblastoma. In predominantly white Caucasian populations the age‐standardized rate was 7–12 per million, and 6–10% of all childhood cancers were neuroblastomas. Rates were highest in the first year of life (25–50 per million, 30% of total neuroblastoma incidence), and decreased with age to 15–20 per million (50% of the total) at age 1–4, 2–4 per million (15%) at 5–9 and 1–1.5 per million (5%) at 10–14. In the United States, black children had an incidence of 8.5 per million compared with 11.5 among Whites; Blacks tended to be older than Whites at diagnosis. The highest rate in Africa was in Bulawayo, Zimbabwe (8.0 per million) and the lowest in West Nile, Uganda, with no cases registered. Incidence in Israel was similar to that in many white populations, with Jews having a particularly high rate. In other parts of West Asia neuroblastoma had a low relative frequency, suggesting that incidence is low. Rates were also low throughout much of southern and eastern Asia, including India and China. Incidence in Japan was somewhat higher, though less than in Western countries, with the deficit most pronounced in the first year of life; these data relate to the period before mass screening of infants for neuroblastoma in the regions concerned. Incidence was generally higher in regions and among ethnic groups enjoying a higher standard of living, though previous studies within single countries had suggested that neuroblastoma is more common among less affluent groups. Blacks in Africa and the United States may have a weaker genetic predisposition to neuroblastoma, but some of the deficit in many developing countries is likely to be due to under‐diagnosis. © 1992 Wiley‐Liss, Inc.
PCR assay for chromosome 1p deletion in small neuroblastoma samples
Authors: Peter Marline , Michon Jean , Vielh Philippe , Neuenschwander Sylvia , Nakamura Yusuke , Sonsino Elise , Zucker Jean‐Michel , Vergnaud Gilles , Thomas Gilles , Delattre Olivier
Published: February 2007

Deletion of the short arm of chromosome I is among the most recurrent cytogenetic alterations found in neuroblastoma and has been associated with short survival. However, this prognostic information, which relies on time‐consuming techniques, is not yet routinely exploited. In order to set up a reliable and simple routine test to determine Ip deletion in neuroblastoma, we have used the polymerase chain reaction to genotype neuroblastoma DNA at 2 loci containing a variable number of tandem repeats and located on the distal part of the short arm of chromosome I. Agarose gel electrophoresis and ethidium bromide staining of the amplification products enable a simple determination of constitutional and tumor genotypes at these loci to be made. A total of 37 samples from 29 patients were studied with this technique. Loss of heterozygosity (LOH) could be identified in 8 cases. In each case the results obtained were in agreement with those achieved by the Southern procedure. This technique will be of particular interest in the pretherapeutic analysis of Ip deletions in small tumor samples obtained by fine‐needle aspirates of the primary tumor. It should also enable retrospective studies from paraffin‐embedded tumor fragments to be made and provide information for the analysis of tumor heterogeneity in neuroblastoma and in other tumors with Ip deletions. © 1992 Wiley‐Liss, Inc.
A population‐based case‐control study of cancers of the nasal cavity and paranasal sinuses in Shanghai
Authors: Zheng Wei , Blot William J. , Shu Xiao‐Ou , Diamond Earl L. , Gao Yu‐Tang , Ji Bu‐Tian , Fraumeni Joseph F.
Published: February 2007

A population‐based case‐control study of cancer of the nasal cavity and sinuses, involving interviews of 60 incident cases and 414 controls, was conducted in Shanghai. Cigarette smoking was associated with a mild elevation in risk of squamous‐cell carcinoma but not cancers of other cell types. Occupational exposures to wood and silica dusts and to petroleum products, and the use of wood and straw as cooking fuel, were linked to moderate increases in risk, while 4‐fold or greater increases were associated with a history of chronic nasal diseases, including those occurring 10 or more years prior to cancer diagnosis. Dietary analyses revealed a significant protective effect of consumption of allium vegetables, oranges and tangerines, with a 50% reduced risk of nasal cancer among individuals in the highest intake group of these foods. Consumption of salt‐preserved vegetables, meat and fish was associated with a significantly increased risk of nasal cancer in a dose‐response fashion, with a 5‐fold excess observed for the heaviest intake of these salted foods. These findings suggest that dietary factors may contribute to the development of nasal cancer. © 1992 Wiley‐Liss, Inc.
Characterization of a mouse‐human chimeric antibody to a cancer‐associated antigen
Authors: Orlandi R. , Figini M. , Tomassetti A. , Canevari S. , Colnaghi M. I.
Published: February 2007

In an attempt to obtain a therapeutic antibody, the murine monoclonal antibody (MAb) MBr 1 (IgM, k), directed against human carcinomas, was converted in a mouse/human chimeric MAb of γ I isotype. The chimeric MAb, γICHI‐MBr 1, retains the ability to specifically bind tumor cells and tissues with no modification in its binding to the normal material tested. γI CHI‐MBr 1 recognizes mucins and high‐molecular‐weight glycoproteins carrying the antigenic determinant and stains a neutral glycolipid extracted from MCF—7 cells. The chimeric and the murine MBr 1 efficiently cross‐inhibit each other on the reference cell line MCF‐7 and the calculated affinity constants amount to 3.8 × 107 and 1.7 × 108M−1, respectively. The human constant region allows γI CHI‐MBr 1 to bind with the FcR on the human monocytic cell line U937 and to efficiently mediate antibody‐dependent cellular cytotoxicity in the presence of human lymphocytes activated by IL2. In addition, γICHI‐MBr 1, like the murine MBr I, mediates complement‐dependent tumor‐cell lysis. Thus, by modelling a molecule with reduced size and increased functional characteristics, we have obtained a reagent which is more suitable for in vivo therapeutic approaches. © 1992 Wiley‐Liss, Inc.
Radiation‐induced lymphoid tumors and radiation lethality are inhibited by combined treatment with small doses of zinc aspartate and wr 2721
Authors: Floersheim G. L. , Christ A. , Koenig R. , Racine C. , Gudat F.
Published: February 2007

Combined small doses of zinc aspartame and WR 2721 provided additive protection against radiation lethality in mice. Survival obtained with a small dose of WR 2721 which was ineffective alone could be enhanced to 83% by combining the drug with zinc aspartate, which on its own also displayed no effect. The survival of 25% provided by a higher dose of WR 2721 was increased significantly by adding zinc aspartate. Additivity was also tested in a model of radiation carcinogene‐sis. For this purpose, lethality and occurrence of lymphoid tumors induced by fractionated total‐body irradiation were studied in C57BI/6 mice treated with zinc aspartate and WR 2721. In order to reveal additive effects, both agents were used at sub‐optimal dosages. In mice subjected to 5 daily exposures of 1.9 Gy, the combination of zinc aspartate and WR 2721 was effective and enhanced the survival to 83% as compared with 25% afforded by WR2721 alone (p < 0.005). Similarly, histological assessment of organ involvement with lymphoma revealed that zinc aspartate and WR 2721 alone did not bring about a significant reduction of lymphoma incidence. On the other hand, the combined agents diminished organ involvement with lymphoma to 9.1 % as against 90% in the controls (p < 0.0005) and 62.5% with WR 2721 alone (p < 0.025). Thus, combined treatment with zinc aspartate and WR 2721 also inhibited radiation‐induced lymphoid tumors. © 1992 Wiley‐Liss, Inc.