bluuebunny/crossref_metadata_2025 huggingface dataset:
Clonal selection within metastatic sp1 mouse mammary tumors is independent of metastatic potential
Authors: Samiei Mitra , Waghorne Carol G.
Published: February 2007

Our previous studies using randomly integrated plasmid DNA as unique clonotypic markers of SP1 mouse mammary tumor cells transplanted into syngeneic CBA/J or nude mice demonstrated reproducible selection and eventual overgrowth of the primary transplant tumors by genotypically distinct metastatic subclones. Two independent metastatic SP1 clones, neo5 and ras1, were shown to exhibit „clonal dominance”︁ relative to the non‐metastatic SP1 tumor‐cell population. These results suggested that the capacity for preferential growth within the tumors may be related to cellular properties associated with metastatic ability. To investigate the clonal interactions of metastatic SPI clones present within the same tumor mass, we have analyzed tumors composed of paired mixtures of neo5 and ras1. The tumors were monitored for the relative proportion of each clone by Southern blot analysis. The ras1 clone was found to dominate over the neo5 clone in the majority of tumors examined, even when present as 1% of the mixed inoculum. This represents a 20‐ to 50‐fold enrichment of ras1, while the proportion of neo5 within the tumors was reduced at least 5‐fold. No evidence for selection of either clone was seen during co‐culture in vitro. Neo5 and ras1 are indistinguishable with respect to tumor igenic and metastatic potential when inoculated separately into different mice, suggesting that clonal dominance is independent of metastatic ability. Analysis of the metastases resulting from mixed inocula indicates that it is possible for a subpopulation representing less than 1% of the primary tumor mass to give rise to metastases. This also suggests that the process of metastasis within metastatic tumors is independent of clonal dominance.
Induction of tumor regression in experimental model of human head and neck cancer by human a‐lak cells and IL‐2
Authors: Sacchi Manuela , Vitolo Domenico , Sedlmayr Peter , Rabinowich Hanna , Johnson Jonas T. , Herberman Ronald B. , Whiteside Theresa L.
Published: February 2007

In a nude mouse model of human squamous‐cell carcinoma of the head and neck (SCCHN), locoregional therapy with interleukin 2 and human lymphokine‐activated killer (LAK) cells resulted in a significant inhibition of growth of 3‐day established tumors. The same model was used for therapy of 7‐day established tumors with highly enriched populations of human adherent (A)‐LAK (CD3− CD56+) cells and IL‐2. Peritumoral transfer of 10 × 106 A‐LAK cells, whose in vitro cytotoxicity against a SCCHN cell line (PCl‐1) was not significantly different from that of LAK cells, resulted in complete regression of all 3‐day or 7‐day human SCCHN in nude mice. An initial inflammatory‐type reaction, which appeared within hours of the first peritumoral cell transfer, was accompanied by infiltration initially by granulocytes and plasma cells, and later by mononuclear cells into the tumor stroma. A‐LAK cells labelled with a fluorescent dye prior to injection appeared in the tumor stroma within 24 hr and were localized around or in the basal epithelial tumor layer by 48 hr. Histologic sections revealed an increasing epithelial disorganization and progressively decreasing basal epithelial layer, which were proportional to the increasing number of A‐LAK cells transferred. Within 4 weeks, the tumors were reduced to amorphous keratinic remnants surrounded by the connective tissue containing abundant mononuclear cells. Local administration of human A‐LAK cells and IL‐2 to SCCHN tumors growing in nude mice led to accelerated tumor differentiation, keratinization and regression.
Biological response of multicellular emt6 spheroids to exogenous lactate
Authors: Bourrat‐Floeck B. , Groebe K. , Mueller‐Klieser W.
Published: February 2007

The influence of elevated lactate concentrations, as found in tumor microregions, on cellular growth, viability, and metabolic state was studied employing the multicellular spheroid model. Spheroids of EMT6/Ro cells were cultured at 37°C in 5% or 20% (v/v) oxygen, using stirred media with various concentrations of exogenous lactate ranging from 0.0 mM (standard conditions) to 20.0 mM. Elevated concentrations of exogenous lactate led to a considerable decrease of the maximum spheroid diameter at growth saturation, e.g., for 20% O2 from around 1700 μm to 700 μm in 0.0 and 20.0 mM lactate respectively. Histological investigations showed that the thickness of the viable cell rim was increased by elevated lactate concentrations in 20% O2, whereas this correlation was reversed in 5% O2. Cultivation of spheroids in increasing lactate concentrations was associated with a shift of metabolic pathways from net production to increased utilization of lactate in both 20% and 5% O2, as determined by standard enzymatic assays. Oxygen tension (PO2) values measured with micro‐electrodes were less in spheroids cultured in high lactate (9.0 and 20.0 mM) than under standard conditions, irrespective of the external oxygen concentration. This finding reflected a substantial increase in the cellular O2 consumption with elevated external lactate levels. At given lactate concentrations, respiration rates that were derived from measured PO2 distributions by theoretical considerations were significantly lower in 5% O2 than in 20% O2.
Characterization of a novel monoclonal antibody, 3H‐1, reactive with squamoproliferative lesions and squamous‐cell cancers
Authors: Parsons Peter G. , Leonard J. Helen , Kearsley John H. , Takahashi Hiroyuki , Lin‐Jian Xu , Moss Denis J.
Published: February 2007

Immunization of mice with membranes from a virus‐transformed human keratinocyte cell line (KJD‐I/SV40) yielded an IgM monoclonal antibody (MAb 3H‐1) which reacted with the membrane and cytoplasm of KJD‐I/SV40 cells and in the perinuclear region of a squamous‐cell carcinoma line (Colo‐16). Immunohistochemistry of formalin‐fixed, paraffin‐embedded sections using MAb 3H‐1 gave intense staining of proliferating squamous epithelium in several characteristic patterns. Acanthotic squamous epithelium and well‐differentiated squamous‐cell carcinomas (SCC) demonstrated a membranous staining pattern whereas psoriatic skin and undifferentiated SCC of the head and neck exhibited diffuse cytoplasmic, focal cytoplasmic or, in some tumours, no staining. Simple squamous epithelium was unreactive. Western blotting revealed an antigen of 55 kDa. The epitope recognized by MAb 3H‐1 may be a marker for particular stages of squamous proliferation and differentiation.
B‐859‐35, a new drug with anti‐tumor activity reverses multi‐drug resistance
Authors: Hofmann Johann , Ueberall Florian , And Alexander Egle , Grunicke Hans
Published: February 2007

It has previously been shown that B‐859‐3S ((–)‐3‐meth‐yl‐5‐ 3‐(4,4‐diphenyl‐1‐piperidinyl)‐propyl‐1,4‐dihydro‐2,6‐dimethyl‐4‐(3‐nitrophenyl)‐pyridine‐3,5‐dicarbox‐ylate‐hydrochloride) exerts a selective carcinostatic effect on some tumors. In order to evaluate whether the anti‐cancer activity of B‐859‐35 can be modulated, we combined the new drug with several established anti‐tumor drugs. A combination of B‐859‐35 with VP‐16 (etoposide) in MDR(multi‐drug‐resistant‐gene)‐expressing Walker rat carcinoma cells shows synergism. A combination of B‐859‐35 with doxorubicin results in stronger synergism than verapamil/doxorubicin, especially at low concentrations of B‐859‐35. The resistance of mdrI (human multi‐drug‐resistance‐gene)‐expressing human HeLa KB‐8‐5 cells to doxorubicin can be reversed with nontoxic or weakly toxic concentrations of B‐859‐35 to the sensitivity of the parent KB‐3‐1 cells. The finding that an antitumor drug is able to reverse multi‐drug resistance makes B‐859‐35 an interesting drug for cancer treatment.
A lack of detectable modification of topoisomerase II activity in a series of human tumor cell lines expressing only low levels of etoposide resistance
Authors: Hill Bridget T. , Whelan Richard D. H. , Hosking Louise K. , Shellard Sharon A. , Hinds Michael D. , Mayes Janice , Zwelling Leonard A.
Published: February 2007

Etoposide (VP‐16) resistance is expressed following in vitro exposure of HN‐1 and MCF‐7 human tumor cells to the drug itself or to fractionated × irradiation. VP‐16‐selected sublines prove cross‐resistant to Adriamycin, amsacrine and actinomycin D, whilst X‐ray‐pretreated sublines show cross‐resistance to only actinomycin D. These differential responses, in the HN‐1 series, are not associated with significant differences in amounts of immunoreactive topoisomerase (topo) II, altered topo‐II catalytic activity of nuclear extracts or changes in susceptibility of the topo II to VP‐16‐ or amsacrine‐induced DNA‐protein cross‐link formation. Therefore significant modifications in topo II appear not to be implicated in VP‐16 resistance in these HN‐1 sublines.
Down‐regulation of E‐cadherin expression in madin darby canine kidney (MDCK) cells inside tumors of nude mice
Authors: Mareel Marc M. , Behrens Jürgen , Birchmeier Walter , De Bruyne Georges K. , Vleminckx Kris , Hoogewijs Annemarie , Fiers Walter C. , Van Roy Frans M.
Published: February 2007

The 120‐kDa cell‐cell adhesion molecule E‐cadherin is localized at the epithelial junctional complex and participates in the organization and maintenance of epithelia. The Madin Darby canine kidney (MDCK) cell line expresses E‐cadherin in a stable way and forms polarized epitheloid structures in vitro. Harvey‐murine‐sarcoma‐virus‐transformed derivatives (MDCK‐ras) produce malignant (i.e., invasive and metastatic) tumors in nude mice. We obtained evidence that E‐cadherin is down‐regulated in nude mouse tumors and that this down‐regulation is reversible. MDCK‐ras‐e cell lines were cloned in vitro from MDCK‐ras cell cultures. They showed an epithelioid morphotype and expressed E‐cadherin at homogeneously high level. This characteristic has been conserved for at least 60 passages in vitro. MDCK‐ras‐e cells were not invasive in vitro. When injected into nude mice, however, they produced invasive and metastatic tumors. Primary tumors as well as large metastases were heterogeneous, showing E‐cadherin‐positive well differentiated epithelial structures and E‐cadherin‐negative undifferentiated areas. Metastasis‐derived cell cultures contained both E‐cadherin‐positive and E‐cadherin‐negative MDCK‐ras‐e cells during early passages in vitro. During further culture, however, they regained the homogeneous E‐cadherin‐positive characteristic of the original MDCK‐ras‐e cell line. The behavior of MDCK‐ras‐e cells in vitro, as compared with its in vivo behavior, points to the existence of host factors which are able to down‐regulate E‐cadherin expression. We hypothesize that this down‐regulation plays a basic role in invasion.
The effect of fetal calf serum on growth arrest caused by activators of protein kinase C
Authors: Bradshaw Tracey D. , Gescher Andreas , Pettit George R.
Published: February 2007

The growth of human‐derived A549 lung carcinoma cells is inhibited by activators of protein kinase C (PKC) such as 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA). In this study, the effect of serum deprivation on TPA‐induced growth retardation has been investigated. Cells cultured with 10% FCS and TPA (10−8 M) stopped growing for 6 days, whereas inhibition of DNA synthesis caused by TPA in cells which were grown in medium containing the serum substitute ultraser lasted for less than 48 hr. The ability of cells to respond to the growth‐inhibitory potential of TPA decreased with decreasing amounts of FCS in the cellular medium. Addition of fetuin or epidermal growth factor (EGF) to incubates with serum‐deprived cells increased the ability of TPA to affect growth, but addition of platelet‐derived growth factor (PDGF), transforming growth factor beta (TGF‐β) or retinoic acid (RA) was without effect. Growth arrest caused by bryostatin I, another PKC activator, was equally transitory in serum‐supplemented and serum‐deprived cells. Cytosol of serum‐deprived cells contained only 32% of specific phorbol ester binding sites compared to cells grown with FCS; PKC enzyme activity and immunodectable protein were similarly reduced in cells grown without FCS. There was no difference in rate of TPA‐induced down‐regulation of PKC activity and cytosolic phorbol ester receptor sites between cells grown with or without serum.
A case‐control study of oral cancer and pre‐diagnostic concentrations of selenium and zinc in nail tissue
Authors: Rogers Mary A. M. , Thomas David B. , Davis Scott , Weiss Noel S. , Vaughan Thomas L. , Nevissi Ahmad E.
Published: February 2007

A case‐control study of oral cancer was conducted in western Washington state between 1983 and 1987. Cases (n = 379) were identified through a population‐based registry, and controls (n = 514) were selected by telephone using random digit dialing. Subjects participated in a personal interview, completed a food‐frequency questionnaire, and submitted clippings from the nails of each great toe for the determination of selenium and zinc concentrations. The odds ratio (OR) for low selenium levels in nail tissue (lowest 25% of the distribution compared to the upper 75%) was 1.4 (95% confidence interval (Cl) 1.0–2.2). Likewise, the odds ratio for low zinc levels in nails was 1.6 (95% Cl 1.0–2.3), but for low dietary zinc was 1.0 (95% Cl 0.7–1.7). Men with oral cancer had lower nail selenium levels than did the controls (OR = 1.9), but women with oral cancer did not (OR = 0.6). Individuals 20 to 39 years of age with oral cancer, in particular, were more likely to have lower selenium levels in nail tissue than controls (OR = 16.4). There was a significant interaction between selenium and ascorbic acid levels which could not be explained by cigarette use. Subjects at greatest risk had low levels of both nutrients (OR = 3.8 for smokers and OR = 5.7 for non‐smokers). However, since the elements were deposited in the nail matrix close to the date of diagnosis, the differences in the element concentrations between cases and controls may have been a result of the disease. Further etiologic studies of selenium, vitamin intake and oral carcinoma are warranted.
Androgen receptors in endocrine‐therapy‐resistant human prostate cancer
Authors: van der Kwast Theodorus H. , Schalken Jack , de Winter Jacobus A. Ruizeveld , van Vroonhoven J. Cor C. , Mulder Eppo , Boersma Wim , Trapman Jan
Published: February 2007

Despite the initial androgen‐dependent growth of most human prostate cancers, eventually all prostate cancers become androgen‐independent at varying intervals after androgen ablation or anti‐androgen therapy. In order to gain more insight into the role of the androgen receptor (AR) in this process, AR and prostate‐specific antigen (PA) expression was evaluated immunohistochemically in prostatic tumour tissues from patients who developed urinary flow obstruction between 4 and 107 months after onset of treatment. AR expression was evaluated with a monoclonal antibody (MAb) specific for the N‐ terminal domain of the human AR. To substantiate the progressive tumour growth, proliferative activity was assessed immunohistochemically by staining with MAb Ki‐67. Ki‐67‐defined tumour‐growth fractions varied from 0.8–64.7%. In 13 of the 17 examined tumours over 80% of the tumour cells were AR‐positive, 3 tumours showed a considerable heterogeneity in AR expression and in I tumour almost all tumour cells seemed to be AR‐negative. Two‐thirds of the examined tumours contained variable proportions of PA‐positive tumour areas. These observations contrast with the view that androgen ablation induces a preferential outgrowth of receptor‐negative tumour cells.
Prognostic significance of argyrophilic nucleolar organizer staining in soft‐tissue sarcomas
Authors: Kuratsu S. , Aozasa K. , Myoui A. , Tsujimoto M. , Ueda T. , Uchida A. , Hamada H. , Ono K. , Matsumoto K.
Published: February 2007

The utility of argyrophilic stain for nucleolar organizer region (AgNOR) for estimating proliferative activity and prognosis of soft‐tissue sarcomas (STS) was examined. Formalin‐fixed and paraffin‐embedded sections of 38 cases with STS were used; the reaction product of AgNOR stain was observed as dots mainly in the nucleoli. The mean number of AgNOR dots per nucleus of tumor cells was calculated in 200 cells (AgNOR count). The AgNOR count, ranging from 1.4 to 16.1 (mean, 7.5), showed a good correlation with cellularity (r = 0.483, p < 0.003) and histologic grade (r = 0.626, p < 0.00005), but less shown with mitotic counts (r = 0.350, p < 0.04). The prognosis of cases with AgNOR low‐count group (5‐year survival rate was 74.6%) was much better than those in high count group (33.3%) (p < 0.0005). The AgNOR count correlated well with reactivity of tumor cells for Ki‐67 staining, which was available only in freshly prepared sections. These findings suggested that the AgNOR staining is a simple and useful method for estimating tumor‐cell proliferation and prognosis of patients with STS.
Transglutaminase activity and Nϵ(γ‐glutamyl) lysine isopeptide levels during cell growth: An enzymic and immunological study
Authors: Alaoui Said El , Legastelois Stéphane , Roch Anne‐Marie , Chantepie Jacqueline , Quash Gérard
Published: February 2007

A monoclonal antibody (MAb) 81D1c2, which recognizes the Nϵ(γ‐glutamyl) lysine isopeptide produced by the action of transglutaminase activity was prepared. Its reactivity towards the homologous isopeptide was about 3‐fold greater than that with either Nα (α glutamyl) lysine (a naturally occuring heterologous dipeptide) or Nϵ(γ glutamyl) lysine, another heterologous peptide not described so far in naturally occuring proteins. When used in an immunohistochemical study on cells in culture derived from human carcinoma of the larynx (HEp2) and from chicken embryo cells (CEC), both fixed in acetone, this MAb detected Nϵ(γ glutamyl) lysine residues In the nucleus. The amount of Nϵ(γ glutamyl) lysine isopeptides follows closely transglutaminase activity during the lag phase of growth of both CEC and HEp2 cells. However, during exponential growth, the 2 parameters decrease concomitantly in HEp2 cells, whereas in CEC, transglutaminase activity increases but isopeptide bond levels drop. Compared with other reported methods for measuring isopeptides, this immunohistological approach permits the localization and at least the semi‐quantitative determination of Nϵ(γ glutamyl) lysine in cells in situ.
V‐ABL does not abolish IL‐6 requirement by murine plasmacytoma cells
Authors: Sugiyama Hiroyuki , Wiener Francis , Babonits Magda , Silva Santiago , Hirano Toshio , Kishimoto Tadamitsu , Klein George
Published: February 2007

Activation of c‐myc by juxtaposition to an immunoglobulin locus and introduction of the v‐ab/ oncogene act synergistically in generating a mouse plasmacytoma (PC). The question arose whether the effect of v‐ab/ could be attributed to a deregulation of interleukin‐6 (IL‐6) production or responsiveness, in view of the fact that IL‐6 exerts potent growth‐stimulatory activity on PC cells. We studied the effect of IL‐6 on the in vitro growth of primary PCs induced by pristane alone (TEPCs) or by pristane + A‐MuLV (ABPCs). Five of 13 TEPCs and 3 of 7 ABPCs responded to IL‐6. Macrophage supernatants prepared from both TEPCs and ABPCs had similar stimulatory effects on PC cells. From 30 primary PCs (including both TEPCs and ABPCs), we established 9 in vitro lines, 2 of which expressed v‐abJ. All were able to grow on macrophage feeder layers. Three types of behavior could be distinguished on the basis of growth in feeder‐free cultures in the presence and absence of IL‐6. Group I contained 4 IL‐6‐dependent lines. Group II contained 2 IL‐6‐independent lines (one v‐abl expressor) that grew faster In the presence of IL‐6. Group III consisted of 3 feeder‐dependent lines (one v‐ab/ expressor) that were not significantly stimulated by IL‐6. These findings indicate that v‐ab/ expression does not Influence IL‐6 dependence or responsiveness by itself. The supernatant of one line in group II was able to stimulate PC cells. All 6 lines of Groups I and II carried a typical (12; 15) translation, while all 3 lines in group III had a variant (6;15) or (15;16) translocation.
Retinoic acid suppression of squamous differentiation in human head‐and‐neck squamous carcinoma cells
Authors: Poddar Susmita , Hong Waun K. , Thacher Scott M. , Lotan Reuben
Published: February 2007

Retinoids (vitamin A analogues) inhibit the squamous differentiation of normal and malignant epithelial cells. This Study investigated the ability of the head‐and‐neck squamous‐cell carcinoma (HNSCC) cell line 1483 to undergo squamous differentiation in the absence and presence of β‐all‐trans retinoic acid (RA). The growth of these cells in culture is accompanied by an increase in keratinocyte trans‐glutaminase, involucrin and keratin KI, 3 established markers of squamous cell differentiation. Higher levels of these differentiation markers were detected in cells cultured in delipidized serum (DLS), from which endogenous retinoids have been extracted, than in cells cultured in fetal bovine serum (FBS), which contains retinoids. Treatment with I μM RA decreased the levels of the various differentiation markers in cells cultured in either FBS or DLS as revealed by labelling of permeabilized cells and by immunoblotting of cell extracts using specific monoclonal or polyclonal antibodies. The cells' ability to cross‐link proteins to form envelopes under the plasma membrane was stimulated in the presence of calcium ionophore but Inhibited by RA. These results indicate that the malignant 1483 HNSCC cells recapitulate the main characteristics of normal squamous‐cell differentiation in culture and that RA suppresses this differentiation as it does in normal keratinizing epithelial cells.
Comparative cytotoxicity of CI‐973, cisplatin, carboplatin and tetraplatin in human ovarian carcinoma cell lines
Authors: Perez R. P. , O'Dwyer P. J. , Handel L. M. , Ozols R. F. , Hamilton T. C.
Published: February 2007

The clinical efficacy of cisplatin‐based chemotherapy for ovarian cancer is frequently compromised by drug resistance or dose‐limiting renal and neurologic toxicities. CI‐973 (NK‐121), a 2‐methyl‐1, 4‐butanediamine analogue of carboplatin, has shown little nephro‐and neuro‐toxicity in pre‐clinical model systems and in phase‐I trials. Its in vitro spectrum of activity against ovarian cancer cell lines has not been previously characterized. The in vitro activities of CI‐973, cisplatin, carboplatin and tetraplatin were compared in several platinum‐sensitive and‐resistant human ovarian carcinoma cell lines. Cytotoxicity was assessed by inhibition of clonogenic survival in soft agar with continuous drug exposure. On a molar basis, cisplatin and tetraplatin were the most potent analogues, while carboplatin was consistently less potent. Cisplatin, carboplatin and CI‐973 elicited a very similar response pattern by Spearman rank correlation, distinct from that seen with tetraplatin. The magnitude of resistance to CI‐973 was comparable to cisplatin in 5 cell lines but was substantially lower in the highly cisplatin‐resistant 2780‐CP70 and OVCAR‐10 cell lines. These results suggest that CI‐973 and teatraplatin may have potential utility in some cases of cisplatin‐resistant ovarian cancer. In addition, our data are consistent with the existence of at least 2 platinum‐resistance phenotypes—one with moderate levels of resistance to cisplatin, carboplatin and CI‐973 but highly resistant to tetraplatin, the other highly resistant to cisplatin and carboplatin but only partially cross‐resistant with tetraplatin and CI‐973. The recognition of different resistance phenotypes may facilitate the study of cellular resistance mechanisms to cisplatin and newer platinum analogues.
Immunological and non‐immunological influence of H‐2Kb gene transfection on the metastatic ability of B16 melanoma cells
Authors: De Giovanni C. , Palmieri G. , Nicoletti G. , Landuzzi L. , Scotlandi K. , Bontadini A. , Tazzari P.‐L. , Sensi M. , Santoni A. , Nanni P. , Lollini P.‐L.
Published: February 2007

The H‐2b‐negative B78HI clone (derived from B16 melanoma) was transfected with the H‐2Kb gene; 4 cell clones expressing membrane H‐2Kb antigens and 2 control clones (transfected with pSV2neo alone) were used for studies of metastatic ability, immunogenicity, NK sensitivity and homo‐typic adhesion. The experimental metastatic capacity of H‐2Kb transfectants In syngeneic mice was greatly diminished in comparison with control and parent cells. Both immune‐mediated and Intrinsic properties of transfectants correlated with their lower metastatic ability. A cell‐mediated cytotoxic response was induced by repeated in vivo immunizations of syngeneic mice followed by in vitro restimulation of effectors when transfectants (but not controls) were used as immunizers and as targets. Moreover, homotypic adhesion of H‐2Kbtransfectants was significantly lower than that of controls. Sensitivity to NK ceils of transfectants was not decreased in comparison to H‐2‐negative controls. It is known that In vitro treatment with IFN‐γ of H‐2‐positive B16 melanoma cells induces a simultaneous increase in H‐2 expression and in experimental metastasis; treatment of H‐2Kb transfectants with IFN‐γ induced a higher Kb expression, but no increase in metastatic ability, thus suggesting that the IFN‐sensitive component that mediates enhancement of metastasis is not H‐2Kb.
Use of combination of monoclonal antibodies directed against three distinct epitopes of a tumor‐associated antigen: Analysis of cell binding and internalization
Authors: Casalini Patrizia , Mezzanzanica Delia , Canevari Silvana , Della Torre Gabriella , Miorn Silvia , Colnaghi Maria Ines , Matzku Siegfried
Published: February 2007

Three monoclonal antibodies (MAbs), MOv17, MOv 18 and MOv19 with tumor‐restricted specificity for human ovarian carcinoma, were tested alone or in double combination with the aim of analyzing their binding and internalization behavior on different In vitro cell lines. Biochemical studies indicated that the 3 MAbs were directed against 3 epitopes of the same 38 kDa surface molecule. By immuno‐electron‐microscopy they exhibited a different internalization behavior since MOv17 Induced evident endocytosis through coated vesicles, whereas MOv18 gave rise to occasional uncoated vesicles and MOv19 was completely unable to promote internalization of the relevant molecule. When tested 2 by 2 there was a binding synergy in one of the 9 possible combinations (125I‐labelled MOv18 and unlabelled MOv19), but no change in the internalization behavior. The binding synergy, which was highly reproducible, was temperature‐dependent and was also evident on glutaraldehyde‐fixed cells. A metabolism involvement is therefore unlikely. This could be attributed to an easier accessibility of the CaMOv18 due to a conformational change of the molecule after MOv19 MAb binding.
Liver cancer in Thailand. I. A case‐control study of cholangiocarcinoma
Authors: Parkin D. Maxwell , Srivatanakul Petcharin , Khlat Myriam , Chenvidhya Dhiraphol , Chotiwan Pornarong , Insiripong Somchai , L'abbé Kristan A. , Wild Christopher P.
Published: February 2007

Potential risk factors for cholangiocarcinoma were investigated in a case‐control study among inhabitants of north‐east Thailand, which included 103 cases from 3 hospitals, with age‐and sex‐matched controls. A clear association with past or present infection with Opisthorchis viverrini, as indicated by raised serum antibodies, was found (o.r. 5.0), and at least two‐thirds of cases can be attributed to this cause. The results suggest that males may be at higher risk than females. There was no association with hepatitis B infection, with aflatoxin Intake as estimated from albumin adducts in serum or with any particular dietary patterns. Alcohol consumption was very low in the population, and the risk associated with regular drinking was non‐significant. Regular users of betel nut—predominantly female—had a high risk (o.r. 6.4), a possible mechanism being through their increased exposure to nitro‐samines.
Liver cancer in Thailand. II. A case‐control study of hepatocellular carcinoma
Authors: Srivatanakul Petcharin , Parkin D. Maxwell , Khlat Myriam , Chenvidhya Dhirapol , Chotiwan Pornarong , Insiripong Somchai , L'Abbé Kristan A. , Wild Christopher P.
Published: February 2007

Potential risk factors for hepatocellular carcinoma were investigated in a case‐control study among inhabitants of north east Thailand. Sixty‐five cases from 3 hospitals, with matched controls, were included. Infection with hepatitis‐B virus was the major risk factor—chronic carriers of HB surface antigen had an estimated relative risk of 15.2. Infection with hepatltis‐C virus appeared to be rare. No increase In risk was found with recent aflatoxin intake, as estimated by consumption of possibly contaminated foods, or by measuring aflatoxin‐albumln adducts in serum. Regular use of alcohol (2 or more glasses of spirits per week) was associated with a nonsignificant elevation in risk (o.r. = 3.4, 95% c.i. 0.8–14.6), but the number of regular drinkers in the population was small. The meaning of an apparent protection conferred by certain food items is uncertain, but a possible role of betel nut in the aetiology deserves further investigation.
Prevalence of antibody against the core protein of hepatitis C virus in patients with hepatocellular carcinoma
Authors: Watanabe Yushiro , Harada Shizuko , Saito Izumu , Miyamura Tatsuo
Published: February 2007

We have cloned the whole structural region of the hepatitis C virus (HCV) genome and transiently expressed the nucleocapsid protein in animal cells. Since the nucleotide sequences of this region of the HCV genome has been shown to be highly conserved among different HCV isolates, the assay detecting the antibody to this expressed protein is useful for studying the pathogenicity of HCV. in this work, we investigated the presence of antibodies to HCV nucleocapsid protein (p22) in patients with hepatocellular carcinoma (HCC) and compared it* frequency with that of antibody to HCV non‐structural protein (C‐100), which is presently applied for blood screening for transfusion and diagnosis for chronic hepatitis C. By a sensitive Western blot analysis, 85 of 102 (83.3%) sera of hepatitis B virus surface antigen (HBsAg)‐negative HCC patients were positive for the antibody to p22 (anti‐p22), whereas 68 of the same 102 cases (66.7%) were positive for the anti‐C100 by ELISA. The prevalence of anti‐p22 in 23 HBV carrier HCC patients, 56 patients with non‐HCC cancer and 100 healthy blood donors were 4.3, 12.5 and 1.0%, respectively. Thus, high prevalence of anti‐p22 in non‐B HCC confirmed that HCV infection is closely related to the development of HCC. Furthermore, the anti‐p22 assay can detect HCV‐infected patients who could not previously be identified as such by the present anti‐C100 assay.
Vegetable and fruit consumption and cancer risk
Authors: Negri Eva , La Vecchia Carlo , Franceschi Silvia , D'Avanzo Barbara , Parazzini Fabio
Published: February 2007

The relationship between cancer risk and frequency of consumption of green vegetables and fruit has been analyzed using data from an integrated series of case‐control studies conducted in northern Italy between 1983 and 1990. The overall dataset included the following histologically confirmed cancers: oral cavity and pharynx, 119; oesophagus, 294; stomach, 564; colon, 673; rectum, 406; liver, 258; gall‐bladder, 41; pancreas, 303; larynx, 149; breast, 2,860; endometrium, 567; ovary, 742; prostate, 107; bladder, 365; kidney, 147; thyroid, 120; Hodgkin's disease, 72; non‐Hodgkin lymphomas, 173; myelomas, 117; and a total of 6,147 controls admitted to hospital for acute non‐neoplastic conditions, unrelated to long‐term dietary modifications. Multivariate relative risks (RR) for subsequent tertiles of vegetable and fruit consumption were derived after allowance for age, sex, area of residence, education and smoking. For vegetables, there was a consistent pattern of protection for all epithelial cancers, with RRs in the upper tertile ranging from 0.2 for oesophagus, liver and larynx to 0.7 for breast. All the trends in risk were in the same direction and significant for all carcinomas except gallbladder. In contrast, no protection was afforded by high vegetable consumption against non‐epithelial lymphoid neoplasms. With reference to fruit, strong Inverse relationships were observed for cancers of the upper digestive and respiratory tract, with RRs in the upper tertile between 0.2 and 0.3 for oral cavity and pharynx, oesophagus and larynx relative to the lowest tertile. The lower the location of the tumour in the digestive tract, the weaker appeared to be the protection afforded. Significant inverse relationships were observed for liver, pancreas, prostate and urinary sites, but not for rectum, breast and female genital cancers or thyroid. No relationship emerged for lymphomas and myelomas. Even in the absence of a clear biological interpretation, the consistency and strength of the patterns observed indicate that, in this population, frequent green vegetable intake is associated with a substantial reduction of risk for several common epithelial cancers, and that fruit intake has a favourable effect, especially on upper digestive cancers and, probably, also on urinary tract neoplasms.
Nutritional factors and pancreatic cancer: A case‐control study from South‐West Poland
Authors: Zatonski W. , Przewozniak K. , Howe G. R. , Maisonneuve P. , Walker A. M. , Boyle P.
Published: February 2007

A population‐based case‐control study of diet and pancreatic cancer has been conducted in the Opole Voivodeship region in Poland, using 110 cases and 195 controls. A full diet history was used to estimate the daily intake of calories, fat, protein, carbohydrate, cholesterol, retinol, vitamin C, and fibre. There Is a strong positive association with the intake of cholesterol with relative risks of 1.90, 3.77, and 4.31 for the 3 upper quartiles compared with the lowest (p, trend = < 0.01). An inverse association was seen with vitamin C intake, with corresponding relative risks of 1.10, 0.30, and 0.37 (p, trend = < 0.01). Weak Inverse associations were seen with the Intake of retinol and fibre. There is evidence of an inverse association with fat intake, particularly unsaturated fats, and of a positive association with the intake of carbohydrate and to a lesser extent, protein. These results further support the potential importance of dietary factors in the aetiology of cancer of the pancreas.
Proteinase‐alpha2 macroglobulin complexes are not increased in plasma of patients with cancer
Authors: Zucker Stanley , Lysik Rita M. , Zarrabi Mohamed H. , Fiore John J. , Strickland Dudley K.
Published: February 2007

α2‐macroglobulin, a major glycoprotein component of plasma, is unique in its capacity to bind and inhibit the proteolytic activities of all classes of proteinases. Since proteinases implicated in cancer dissemination (type‐IV collagenase, plasminogen activator, cathepsins B) are normal constituents of blood, we have explored the hypothesis that elevated tissue levels of activated proteinases bound to α2M might be detected in plasma of patients with cancer. To test this premise, blood was collected from 149 subjects (33 healthy controls, 31 patients with infections and non‐malignant diseases, 16 with myeloproliferative disease, 10 with gastrointestinal cancer, 7 with genito‐urinary cancer, 16 with lung cancer, 14 with lymphoma, 11 with miscellaneous cancers and 11 with chronic lymphocytic leukemia and myeloma). Plasma was assayed for α2M‐proteinase complexes using a sandwich ELISA which employs a mouse monoclonal antibody (MAb) that binds to a neo‐antigenic determinant on complexed α2M and a rabbit polyclonal anti‐native human α2M antibody. The concentration of complexed α2M in healthy controls was 14.2 ± 9.8 μg/ml (mean ± standard deviation). No significant differences in complexed α2M were noted between normal and cancer groups (range 7.4–14.6 μg/ml). On the basis of these data, we propose that, in patients with cancer, activated proteinases are bound locally to inhibitors in the tissues and are not available to form complexes with plasma α2M. An alternative explanation is that proteinases are not secreted in excess by cancer cells in vivo.
Difference in prevalence of human papillomavirus genotypes in cytomorphologically normal cervical smears is associated with a history of cervical intraepithelial neoplasia
Authors: Van Den Brule Adriaan J. C. , Walboomers Jan M. M. , Maine Marc Du , Kenemans Peter , Meijer Chris J. L. M.
Published: February 2007

The prevalence of human papillomavirus (HPV) genotypes was investigated by the pofymerase chain reaction (PCR) method in cytologically normal and abnormal cervical scrapes obtained from asymptomatic women (n = 1,346), participating in a triennial screening program for cervical cancer, and from a gynecological outpatient population (n = 593). In the symptom‐free population oncogenic HPV types 16, 18, 31 and 33 were present in 1.5% of cytologically normal scrapes, while the overall HPV prevalence rate was 3.5%. Significantly, higher HPV prevalence rates of 7% (oncogenic HPV; p < 0.01) and 14% (all HPV; p < 0.01), respectively, were found in cytologically normal scrapes of the gynecologic outpatient population. It appeared that in this outpatient group 78% of the smears containing HPV 16 and 18 were associated with a history of cervical pathology, i.e. cervical intraepithelial neoplasia grade I to III. In smears with mild and severe dysplasia and smears suspected of carcinoma in situ from both populations, the overall HPV prevalence was 70%, 84% and 100%, respectively. In all squamous‐cell carcinomas of the cervix (n = 50) HPV was detected. Frequencies of HPV 16 and 18 increased from 41% in mild dysplasia to 94% in cervical carcinomas. Since a low prevalence of HPV was found in cytomorphologically normal cervices of women without a clinico‐pathological history, the findings in this study suggest that HPV detection in population‐based screening programs for cervical neoplasia can be an important tool in identifying women who are at risk of developing dysplasia and cervical cancer.
Carcinoembryonic antigen regulation in human colorectal tumor cells by a site‐selective cyclic amp analogue: A comparison with interferon‐gamma
Authors: Guadagni Fiorella , Tortora Giampaolo , Roselli Mario , Clair Tim , Cho‐Chung Yoon Sang , Schlom Jeffrey , Greiner John W.
Published: February 2007

Treatment of human colorectal tumor cells (LS174T, HT29, and WIDr) with analogues of cyclic AMP (cAMP) (dibutyryl‐cAMP and 8‐Cl‐cAMP) selectively enhances the expression of carcinoembryonic antigen (CEA). Dose and temporal kinetics results revealed that 8‐Cl‐cAMP was approximately 100‐fold more potent than dibutyryl‐cAMP for increasing CEA expression. Results demonstrated that 8‐Cl‐cAMP treatment of LS174T quantitatively increased CEA levels in cell extracts 2‐fold, increased anti‐CEA monoclonal antibody (MAb) binding to the tumor cell surface, and induced the appearance of CEA‐related mRNA transcripts. The findings suggest that 8‐Cl‐cAMP is capable of regulating CEA expression at transcriptional and/or post‐transcriptlonal levels. Other human tumor cells, as well as normal cell types which do not constitutively express CEA, remained CEA‐negative following 8‐Cl‐cAMP treatment. Moreover, the level of expression of other human tumor antigens as well as antigens of the major histocompatibility complex were not changed by 8‐Cl‐cAMP treatment, suggesting some selectivity for CEA regulation by this cAMP analogue. In vivo administration of 8‐Cl‐cAMP to athymic mice bearing LS174T tumor xenografts increased the amount of anti‐CEA MAb bound to tumor extracts as well as the tumor localization of a radlonuclideconjugated anti‐CEA MAb. The results indicate that 8‐Cl‐cAMP can selectively upregulate CEA expression on human colorectal tumor cells in vitro and in vivo. Interestingly, IFN‐γ treatment of the LS174T cells falls to enhance or induce expression of CEA or any of the histocompatibility leukocyte antigens. Thus, 8‐Cl‐cAMP treatment regulates CEA expression through another cellular pathway which may involve cAMP‐dependent protein kinase.
Analysis of abnormal expression of G‐CSF gene in a novel tumor cell line (KHC 287) elaborating G‐CSF, IL‐1 and IL‐6 with co‐amplification of c‐myc and c‐ki‐ras
Authors: Suzuki Akira , Takahashi Takayuki , Okuno Yoshiaki , Nakamura Kishiko , Tashiro Hironori , Fukumoto Manabu , Konaka Yoshiteru , Imura Hiroo
Published: February 2007

We established a human carcinoma cell line (KHC 287) from a patient with large‐cell‐type lung carcinoma associated with marked leukocytosis. The culture supernatant of KHC 287 cells promoted granulocytic colony formation of human bone‐marrow cells in semi‐solid culture. Colony formation was almost completely suppressed by treatment of the supernatant with a monoclonal anti‐granulocyte colony‐stimulating factor (G‐CSF) antibody. Not only G‐CSF but also interleukin‐la (IL‐1 a), IL‐Iβ and IL‐6 were detected in the culture supernatant by an ELISA method. Northern blot analysis of KHC 287 cells revealed distinct expression of these cvtokine genes. Southern blot hybridization of KHC 287 DNA snowed 20‐ and 40‐fold co‐amplification of c‐myc and c‐ki‐ras, respectively. The chloramphenicol acetyl transferase (CAT) activity was distinctly enhanced in the KHC 287 cells which were transfected with the 360 bp upstream region of G‐CSF gene inserted into pSVOOCAT, but not in non‐G‐CSF‐producing tumor cell lines. These results suggest that overproduction of the transactivating factor(s) which binds to the 360 bp of the G‐CSF upstream region is responsible for the abnormal expression of G‐CSF gene in KHC 287 cell line.
New stable butyrate derivatives alter proliferation and differentiation in human mammary cells
Authors: Planchon P. , Raux H. , Magnien V. , Ronco G. , Villa P. , Crépin M. , Brouty‐Boyé D.
Published: February 2007

Two new butyric esters which were devised to extend the half‐life of n‐butyric acid in vivo, were used to study their effects on a number of phenotypic characteristics including cell morphology, cell proliferation, colony formation, cell‐surface antigen and estrogen receptor expression in 3 normal immortalized cell lines and 2 carcinoma cell lines derived from the human mammary gland. When treated with butyric esters, human mammary cells acquired numerous cytoplasmic granules and vacuoles, reminiscent of secretory functions, and increased In volume. Modulation of the expression of membrane‐associated antigens recognized by the monoclonal antibodies (MAbs) 115D8, 140C1 and 125B5 was also observed. Furthermore, butyrate derivatives inhibited the proliferation of all the cell lines tested and the colony‐forming capacity of those that grew in soft agar. The inhibitory effects were, however, reversible upon removal of butyric esters from the culture medium. In the human breast carcinoma cell line, MCF‐7, in which the cytostatic effects of butyric esters were the most pronounced, cells accumulated in the G0/G1 phase of the cell cycle. This cell line was the only one to contain estrogen receptors which decreased in number when treated with butyric esters without any modification in their binding affinity. Moreover, the stimulatory effects of estrogen on MCF‐7 cell proliferation were antagonized by butyric esters. Our results demonstrate that many of the proliferative and differentiation changes previously reported for n‐butyrates in tumor cells are similarly produced by the new stable butyrate derivatives in normal and malignant human mammary cell lines.
Diet, alcohol consumption and reproductive factors in a case‐control study of breast cancer in Moscow
Authors: Zaridze David , Lifanova Yelena , Maximovitch Dmitrii , Day Nicholas E. , Duffy Stephen W.
Published: February 2007

A case‐control study was conducted in Moscow to assess the effect of diet on risk of breast cancer, and also to study the established reproductive risk factors. A notable finding of the study, which covered 139 case‐control pairs matched by age and neighbourhood, is that dietary factors are more important for post‐menopausal than for pre‐menopausal breast cancer. The decreased risk of post‐menopausal breast cancer was associated with high intakes of cellulose (OR 0.04; 95% CI 0.01–0.31), mono‐ and disaccharides (OR 0.02; 95% CI 0.002–0.27), vitamin C (OR 0.20; 95% CI 0.06–0.70), beta‐carotene (OR 0.09; 95% CI 0.02–0.49), and also polyunsaturated fatty acids (OR 0.14; 95% CI 0.03–0.69). High intakes of total fat resulted in a statistically non‐significant decrease in the odds ratio (OR 0.52; 95% CI 0.04–6.99), while saturated fats slightly increased the risk of breast cancer (OR 1.67; 95% CI 0.24–11.78). Protein intake was also associated with increased risk of breast cancer (OR 4.62; 95% CI 0.69–30.70). Alcohol use significantly increased the risk of breast cancer in post‐menopausal women (OR 3.39; 95% CI 1.37–4.38). In general, the results of our study indicate that high risk of breast cancer is associated with high intakes of nutrients derived from animal products, and low risk with high intake of those from vegetables and fruits.
Cloning and characterization of human papillomavirus type 52 from cervical carcinoma in Indonesia
Authors: Takami Yasunari , Kondoh Gen , Saito Junko , Noda Kiichiro , Sudiro Tjahjani Mirawati , Sjahrurachman Agus , Warsa Usman Chatib , Yutsudo Masuo , Hakura Akira
Published: February 2007

Human papillomavirus (HPV) type 52 from a cervical carcinoma in Indonesia was molecularly cloned and characterized. By hybridization with cervical carcinoma DNAs from Indonesian patients, HPV 52 was detected in 3 of 52 cases (6%), whereas HPV 16 and 18 were detected in 8 and 7 cases, respectively (15% and 13%). Sequence analysis revealed that the E6‐E7 ORFs contained several DNA binding motifs (Cys‐X‐X‐Cys) like previously sequenced HPVs. The E6 ORF also contained splice donor and acceptor signals, which may allow the expression of E6* protein. The E7 ORF encoded an amino acid sequence that is conserved in some DNA tumor viruses and is involved in binding to Rb protein and in cellular transformation. Transfection of a subgenomic fragment of HPV 52 under the control of a heterologous promoter showed that the E7 ORF alone induced anchorage‐independent growth of established rodent cells and immortalized primary rat embryo fibroblasts (REF), and that In cooperation with activated ras, it induced malignant transformation of REF. The E6 ORF also induced, less efficiently, anchorage‐independent growth. These results strongly suggest that HPV 52, like HPV 16 and 18, has oncogenic potential.