bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro activity of novel antifolates against human squamous carcinoma cell lines of the head and neck with inherent resistance to methotrexate
Authors: van der Laan B. F. A. M. , Jansen G. , Kathmann G. A. M. , Westerhof G. R. , Schornagel J. H. , Hordijk G. J.
Published: February 2007

A series of 7 human squamous carcinoma cell lines of the head and neck (HNSCC), grown in standard medium containing high folate concentrations and in “folate‐conditioned” medium containing nanomolar concentrations of folates, were all found to be sensitive (IC50: ⩽50 nM) in growth‐inhibition studies to methotrexate (MTX) following drug exposure for 7 days. However, when MTX exposure was limited to 24 hr, only 2 out of 7 HNSCC cell lines were sensitive to MTX (IC50: <500 nM), 2 were moderately sensitive (IC50: 1‐2 μM), and 3 exhibited inherent resistance to MTX (IC50: >250 μM). In these last 3 cell lines, the mechanism of resistance was not correlated with altered membrane transport of MTX or changes in dihydrofo‐late reductase activity, but rather was associated with a 3‐fold lower activity of intracellular folylpolyglutamate synthase (FPGS) activity compared to MTX‐sensitive HNSCC cells. The 3 cell lines exhibiting inherent resistance to a short exposure to MTX, however, did not show inherent cross‐resistance after exposure for 24 hr to one or more of 3 novel antifolate compounds. These compounds, which appear to be more efficiently transported and polyglutamylated than MTX, include: 10‐ethyl‐10‐deazaami‐nopterin (10‐EdAM), 2‐desamino‐2‐methyl‐N10‐propargyl‐5,8‐dideazafolic acid (ICI‐198,583), and 5, 10‐dideazatetrahydrofo‐lic acid (DDATHF). These results indicate that antifolate membrane transport and intracellular FPGS activity are important factors in determining sensitivity or resistance of HNSCC cells to short‐term antifolate compound exposures.
Physiological factors influencing radioantibody uptake: A study of four human colonic carcinomas
Authors: Blumenthal Rosalyn D. , Sharkey Robert M. , Kashi Rina , Natale Ana M. , Goldenberg David M.
Published: February 2007

We evaluated the accretion of 131l‐labeled NP‐4 anticarcinoembryonic antigen (CEA) into 4 size‐matched human colonic carcinomas grown s.c. in nude mice. Antibody uptake for LS174T and GW‐39 tumors was relatively high (19 to 23%ID/g on day 3), whereas moderate uptake was seen in the Moser tumor (7.5% on day 3) and low uptake was detected in the GS‐2 tumor (1.8% on day 3). Blood clearance of radioanti‐body was twice as fast in mice with GS‐2 tumors than in mice with GW‐39, LS174T or Moser tumors. Seven physiological parameters that might influence radioantibody accretion were evaluated in order to better understand the differences in observed tumor targeting: vascular volume, blood flow rate, vascular permeability, tumor antigen content, serum antigen content and complexation of radioantibody, intratumoral antigen distribution, and intracellular antigen distribution. Although marked variability in vascular physiology, antigen content and antibody complexation of the 4 tumors grown in the same host and site existed, it was insufficient to explain the differences in antibody uptake. However, intra‐tumoral distribution of antigen, and sub‐cellular accessibility of antigen for radioantibody were important considerations. GS‐2 tumors are well differentiated and have polarized cells. CEA in GS‐2 is largely inaccessible to radioantibody; most of the antigen is located in the lumen of the glands or on the apical surface of gland cells and most of the antibody distributes to the stromal region on the basolateral surface. The low antibody targeting in GS‐2 could therefore be explained by restricted intra‐tumor accessibility of antibody. Scatchard analysis of NP‐4 binding to Moser cells under non‐internalizing and internalizing conditions revealed that 90% of the antigen is found within the cell, unavailable to bind with the NP‐4 antibody, which is slow to internalize. In contrast, CEA in LS174T cells was almost entirely accessible. The reduced antibody targeting to Moser xenografts might therefore, be explained by restricted antibody accessibility at the cellular level.
Therapy of primary and metastatic mouse mammary carcinomas with doxorubicin encapsulated in long circulating liposomes
Authors: Vaage Jan , Mayhew Eric , Lasic Dan , Martin Frank
Published: February 2007

The purpose of our study was to compare the therapeutic effects of doxorubicin in 3 different formulations: (1) in PBS, (2) in conventional liposomes composed of egg phosphatidylglycerol/ egg phosphatidylcholine/cholesterol/dl‐α tocopherol, and (3) in sterically stabilized, long‐circulating “Stealth” liposomes composed of hydrogenated soy phosphatidylcholine/cholesterol/ polyethylene glycol‐distearoylphosphatidylethanolamine. The doxorubicin formulations were used to treat recently implanted and well‐established, growing primary mouse mammary carcinomas, and to inhibit the development of spontaneous metastases from intra‐mammary tumor implants. In the treatment of recently implanted primary tumors, the formulations were given in 3 i.v. injections over 15 days, starting 3 or 10 days after tumor implantation. In the treatment of well‐established primary tumors, the mice received 4 i.v. injections over 22 days, starting an average 38 days after tumor implantation. In the preventive treatment against metastases, the formulations were given in 4 i.v. injections over 22 days, starting 22 days or 58 days after primary tumor implantation. The Stealth liposome formulation was significantly more effective than the conventional liposome formulation or the free drug in reducing the incidence of metastases from intra‐mammary implants of tumor MC19 and tumor MC65, in curing mice with recent implants of tumor MC2A, tumor MC2B, and tumor MC65, and in increasing the 8‐week survival of mice with well‐established implants of tumor MC2B. It is concluded that the long circulation time of the Stealth liposome doxorubicin formulation accounts for its superior therapeutic effectiveness. © 1992 Wiley‐Liss, Inc.
Interferon‐α/β in virus‐induced mouse mammary carcinogenesis: Effects on the spontaneous process and on the progression of transplanted pre‐neoplastic lesions
Authors: Basolo F. , Fontanini G. , Serra C. , Dolei A. , Proietti E. , Belardelli F. , Conaldi P. G. , Bistocchi M. , Squartini F. , Toniolo A.
Published: February 2007

Low levels of anti‐viral activity, mainly interferon α/β (IFN‐α/ β), are regularly found in lymphoid tissues of BALB/c mice infected with the C3H strain of mammary tumor virus. At the time of tumor development, significant amounts of anti‐viral activity were detected in homogenates of spleen and mammary tumors, but not in blood and normal mammary glands. This activity is pH2‐resistant and neutralized by antibody to IFN/ α‐β. The pathogenetic role of IFN in mammary carcinogenesis was investigated in 2 ways: (a) by treating virus‐infected newborn mice with antibody to IFN‐α/β, and (b) by giving either the latter antibody or IFN‐α/β to virus‐free animals transplanted with pre‐neoplastic lesions. Mice were treated only for 2 months, starting either I week after birth or immediately after tumor transplant. In case (a), treatment with antibody to IFN‐α/β shortened the incubation period of mammary carcinomas and decreased the mean survival time. In case (b), anti‐IFN antibody did not significantly affect the development of mammary tumors. However, exogenous IFN‐α/β markedly reduced both tumor incidence and mortality rate. These results indicate that endogenous IFN‐α/β plays a crucial role in the in vivo restriction of the early infectious phase of spontaneous carcinogenesis and that relatively high doses of IFN‐α/β may inhibit the progression of pre‐neoplastic lesions. © 1992 Wiley‐Liss, Inc.
Optimization of culture conditions for activation and large‐scale expansion of human T lymphocytes for bispecific antibody‐directed cellular immunotherapy
Authors: Lamers C. H. J. , van de Griend R. J. , Braakman E. , Ronteltap C. P. M. , Bénard J. , Stoter G. , Gratama J. W. , Bolhuis R. L. H.
Published: February 2007

We investigated the optimal culture conditions (i.e., activation procedure, medium composition and type of culture vessel) for rapid in vitro expansion of large numbers (>5 × 109) of blood T lymphocytes. These expanded lymphocytes can be targeted to be cytotoxic to ovarian carcinoma cells with a bispecific monoclonal antibody (BsAb) specific for CD3 and for the ovarian carcinoma‐associated antigen MOvl8. Both phytohemagglutinin (PHA) and monoclonal antibody (MAb) CD3 induced rapid T‐cell proliferation, although the growth kinetics after PHA activation were slightly faster. A 50‐fold increase in cell number was obtained after 14 and 16 days for PHA and CD3 MAb, respectively. The induction of BsAb‐directed cytolysis was faster after CD3 MAb than after PHA activation of lymphocytes, but became similar around day 20. A mixture of media consisting of 78% RPMI 1640, 20% AIM‐V and 2% human plasma (Mix‐med) yielded better results than 100% AIM‐V medium. Culture of lymphocytes in polyolefin bags, compared with tissue culture flasks, or cryopreservation did not affect lymphocyte yield and function. In most cultures the proportion of CD8+lymphocytes increased, suggesting a growth advantage of CD8+over CD4+ lymphocytes in this culture system. A protocol employing PHA activation, Mix‐med and polyolefin bags has been used successfully to activate and expand blood lymphocytes for the first 5 patients entered into a phase‐l/ll clinical trial for the intraperitoneal treatment of ovarian carcinoma using CD3 x anti‐MOv 18 BsAb‐directed T lymphocytes. © 1992 Wiley‐Liss, Inc.
Expression of histo‐blood‐group‐A/B‐gene‐defined glycosyltransferases in normal and malignant epithelia: Correlation with A/B‐carbohydrate expression
Authors: Mandel Ulla , Langkilde Niels C. , Osrntoft Torben F. , Therkildsen Marianne Hamilton , Karkov Jens , Reibel Jesper , White Thayer , Clausen Henrik , Dabelsteen Erik
Published: February 2007

Malignant transformation of oral and bladder epithelia is often associated with loss of histo‐blood‐group‐A‐ and ‐B‐carbohydrate antigens, whereas these antigens, which are absent in normal adult distal colon (but present in fetal colon) reappear in malignant distal colon. In order to gain insight into the genetic basis of the biosynthetic regulation for these changes, we have correlated the expression of the A‐ and B‐carbohydrate antigens with that of the A/B‐gene‐defined glycosyltransferases in colon, bladder and oral carcinomas by immunohistology. A newly developed anti‐A/B‐transferase monoclonal antibody (MAb) was used to demonstrate the in situ localization of transferase expression at the individual cell level with correlation to carbohydrate antigen expression, and gave the essential information that the transferase is derived from the ABO gene complex. The reappearance of A‐ and B‐carbohydrate antigens in carcinomas of the distal colon was found to be unrelated to the expression of the A/B‐transferees proteins, which were expressed throughout normal adult colon in accordance with previous enzymatic studies. In contrast, the loss of A‐ and B‐carbohydrate antigens in malignant bladder and oral epithelia was accompanied by concordant loss of enzymes. © 1992 Wiley‐Liss, Inc.
Expression of p53 protein in cutaneous melanoma
Authors: Akslen Lars A. , Mørkve Odd
Published: February 2007

The purpose of the present study was to examine the expression of p53 protein in malignant melanomas of the nodular sub‐type, with special reference to the role of p53 alterations in the development of metastases. Increased expression of p53 protein was found in 29 of 30 cases using PAb 1801 monoclonal antibody (MAb) and flow‐cytometric detection on archival material. A large proportion of the cells were positive in a majority of the cases. The level of expression was not correlated to DNA ploidy or to subsequent development of local metastases. However, a significant decrease in p53 protein expression was observed in metastatic lesions, as compared with the corresponding primary tumors. Our results indicate that p53 alterations may be an early event in melanoma development, since almost all cases were affected, and that metastatic spread appears to be parallelled by decreased p53 protein expression. © 1992 Wiley‐Liss, Inc.
Aspects of medical history and exocrine carcinoma of the pancreas: A population‐based case‐control study in The Netherlands
Authors: De Mesquita H. B. Bueno , Maisonneuve P. , Moerman C. J. , Walker A. M.
Published: February 2007

During 1984–88 a population‐based case‐control study was carried out in The Netherlands, in collaboration with the International Agency for Research on Cancer, to examine the possible relationship between aspects of medical history and exocrine pancreatic carcinoma in 176 cases and 487 controls. About 58% of patients were interviewed directly. We observed an inverse relationship between medical treatment for allergy‐ related conditions and the development of pancreatic cancer (30 cases vs. 130 controls, OR 0.57, 95% Cl 0.36 to 0.90). A history of gallbladder problems, gallstones, cholecystectomy, stomach or duodenal ulcer, pancreatitis, appendicitis, diabetes or tonsillectomy was not related to risk. In direct responses, compared with once daily, a positive relationship was seen for stool frequency, 10 years ago, of less than once daily (18 cases vs. 40 controls, OR 2.10, 95% Cl 1.09 to 4.04). In men, diabetes treated with insulin and diagnosed more than 1 year previously was significantly and positively related to risk (5 cases vs. 1 control, OR 11.66, 95% 1.28 to 105.95). In brief, the results of the present study suggest that a history of allergy‐related conditions may protect, whereas a past stool frequency of less than once daily may enhance the risk of cancer of the pancreas. Other elements of the medical history were not consistently related to risk. © 1992 Wiley‐Liss, Inc.
Anthropometric and reproductive variables and exocrine carcinoma of the pancreas: A population‐based case‐control study in The Netherlands
Authors: De Mesquita H. B. Bueno , Maisonneuve P. , Moerman C. J. , Walker A. M.
Published: February 2007

During 1984–88 a population‐based case‐control study was carried out in The Netherlands, in collaboration with the International Agency for Research on Cancer, to examine the possible relationship between aspects of medical history and exocrine pancreatic carcinoma in 176 cases and 487 controls. About 58% of patients were interviewed directly. In women, a significant, positive dose‐response effect of height was seen (p‐value trend < 0.005). Compared with ages 14 or more, women with an early age at menarche, i.e., 11 years or less, had a 3‐fold increase in risk (15 cases vs. 23 controls, OR 3.07, 95% Cl 1.35 to 7.00). Other aspects of the reproductive history were not related to risk. In brief, the results of the present study support the hypothesis that, in women, early menarche and greater adult stature may be early predictors of the development of cancer of the pancreas later in life. © 1992 Wiley‐Liss, Inc.
Immunoreactivity, pharmacokinetics and bone marrow dosimetry of intrathecal radioimmunoconjugates
Authors: Moseley R. P. , Papanastassiou V. , Zalutsky M. R. , Ashpole R. D. , Evans S. , Bigner D. D. , Kemshead J. T.
Published: February 2007

Ten patients with neoplastic meningitis were treated with a variety of 1311‐monoclonal antibody (MAb) conjugates, chosen to bind to their particular malignancy. Pharmacokinetic studies revealed that MAbs leave the ventricular compartment, enter the sub‐arachnoid space and then pass into the blood. Once the MAbs enter the blood compartment, their clearance is determined by factors such as circulating anti‐mouse Ig and circulating antigens. These lead to complex formations and the clearance of the conjugate by the ireticuloendothelial system. In one individual, the anti‐mouse Ig response observed systemically was not mirrored within the CSF, which has implications for planning future therapy of this type. In other patients, formation of immune complexes was due to binding to circulating antigen within the blood. The major toxicity associated with the intrathecal administration of 1311‐MAbs was bone‐marrow suppression. The doses to the bone marrow, resulting from the form of therapy, were calculated but showed no direct correlation with WHO grade 3/4 toxicity. Doses to the ventricular lining werct also calculated, but due to the complex geometry of the compartment, calculation of potential tumour doses was not practicable. © 1992 Wiley‐Liss, Inc.
Expression of genes related to the human erbB, erbA, pdgf and pdgf‐r in tumors of different etiology in Xiphophorus
Authors: Zechel C. , Peters H. , Schleenbecker U. , Anders F.
Published: February 2007

The melanoma determining; Tu locus of the teleost Xiphophorus contains an accessory gene, x‐erbB*a, which is closely related to the EGF receptor gene family, and is probably oncogenic. x‐erbB*a exists in allelic forms that are specific for distinct Tu‐loci, and shows high homology to a non‐allelic non‐oncogenic counterpart x‐erbB*i which is transcribed into mRNA of 4.6 kb in non‐tumorous and tumorous tissues of fish harboring and lacking Tu. Expression of a 4.0‐kb mRNA in tumors (melanoma and fibrosarcoma) of different etiology is strictly correlated with the inheritance of X. maculatus x‐erbB*a alleles; transcripts of 8.0 kb were detected in melanoma and carcinoma of fish harboring a certain x‐erbB*a of X. variatus. The expression of the putative x‐erbB*a transcripts parallels the stage of malignancy of the tumor. The expression of the xiphophorine EGF receptor gene (x‐erbB) was detected in almost all tumors, is strongly enhanced in carcinoma, and is positively correlated with the degree of malignancy of melanoma and fibrosarcoma. Some tumors show expression of erbA‐related genes. The PDGF receptor mRNA is expressed in all tumors analyzed and shows enhanced expression in malignant tumors of neurogenic, epithelial and mesenchymal origin. Expression of x‐pdgf was observed in several cases of melanoma, but more frequently in carcinoma and fibrosarcoma. We conclude that x‐erbB*a might be involved in initiation of tumors of different cellular origin and etiology in fish harboring Tu, as well as in the determination of the malignancy of the tumor. Furthermore, we assume that x‐erbB*i, x‐erbB, x‐pdgf and x‐pdgf‐r play a role in secondary events in tumorigenesis by, e.g., conferring a selective growth advantage to the tumor cells. © 1992 Wiley‐Liss, Inc.
Differentiation‐related expression of adhesion molecules and receptors on human neuroblastoma tissues, cell lines and variants
Authors: Gross Nicole , Favre Simone , Beck Daniel , Meyer Manuela
Published: February 2007

The expression of cellular adhesion molecules (CAM) involved in cell adhesion and immune recognition was measured on neuroblastoma tissue samples, on a neuroblastoma (NB) cell line, SK‐N‐SH, and on 3 phenotypically different variants, SH‐SY5Y, SH‐EP, SH‐IN, representing neuronal, Schwannian/ glial or intermediate NB‐cell types. Immunohistochemical analysis of CAM expression by NB and related tumors at different stages of differentiation revealed a co‐expression of several CAM (ICAN‐1/CD54, LFA‐3, VLA‐2 and HLA‐ABC) associated with low stages and more highly differentiated NB tumors and peripheral neuroepitheliomas (PN). In contrast, N‐CAM was uniformly expressed on all NB tumors. Flow cytometric analysis of CAM surface expression by SK‐N‐SH and variant cells revealed highly variable phenotypes. Expression of ICAM‐1, LFA‐3, VLA‐2 and HLA‐ABC molecules was associated with the epithelial cell type represented by the SH‐EP variant. In contrast, low expression of these molecules and high expression of N‐CAM was associated with the neuronal SH‐SY5Y cells. Exposure of the NB cells to differentiation inducers (retinoic acid, 5′‐brornodeoxyuridine and phorbol esters) and cytokines (τ‐interferon, α‐tumor necrosis factor) resulted in a variable up‐regulation of the expression of all CAMs, except N‐CAM, regardless of the type of differentiation induced. In an attempt to establish a link between the pattern of expression of CAM on NB cells and their susceptibility to natural killer (NK) or lymphokine‐activated killer (LAK) cell lysis, the analysis revealed that NB cells expressing CAM and a differentiated phenotype were less susceptible to NK lysis, but no difference in the sensitivity of the NB cell types to LAK effectors was observed. Treatment of NB target cells with cytokines or PMA decreased their susceptibility to NK and LAK lysis, while induction of differentiation with RA or BUdR resulted in no changes in the sensitivity to NK and LAK lysis. In conclusion, expression of HLA‐ABC and several co‐regulated CAMs was shown to be associated with a differentiated phenotype in NB, with an overall decreased sensitivity to NK/LAK effector cells. © 1992 Wiley‐Liss, Inc.
Orthotopic and ectopic organ environments differentially influence the sensitivity of murine colon carcinoma cells to doxorubicin and 5‐fluorouracil
Authors: Wilmanns Christoph , Fan Dominic , O'Brian Catherine A. , Bucana Corazon D. , Fidler Isaiah J.
Published: February 2007

We determined the effects of organ environment on the response of murine CT‐26 colon carcinoma cells to 2 structurally and pharmacologically distinct chemotherapeutic agents. CT‐26 cells were injected i.v. (to produce lung lesions), s.c., into the cecal wall, and into the spleen (to produce spleen and liver lesions). Doxorubicin (DXR) at 10 mg/kg, 5‐fluorouracil (5‐FU) at 20 mg/kg, or saline (control) was injected intravenously on different schedules after tumor‐cell implantation. The in vivoresponses of the tumors growing in the cecum, spleen, liver, lung and subcutis were compared. Colon carcinomas growing in the subcutis were most sensitive to DXR. Tumors growing in the spleen and cecum were most sensitive to 5‐FU and less so to DXR. Tumors in the liver were highly resistant to both drugs, whereas experimental lung metastases were sensitive to 5‐FU but resistant to DXR. The differential responses of the tumors to the drugs were not due to drug distribution. The level of protein‐kinase‐C activity was elevated in the spleen, liver and cecum tumors as compared with s.c. tumors and correlated with the in vivo DXR resistance of the tumor cells. This correlation suggested that organ environment may modulate the chemosensitivity of tumor cells, at least in part, by perturbing signal transduction pathways. Collectively, the data indicate that the organ environment has profound effects on the response of tumor cells to chemotherapy. A molecular understanding of this phenomenon should facilitate the design of more effective systemic chemotherapy for cancer metastases. © 1992 Wiley‐Liss, Inc.
Secretion of autocrine growth‐promoting activity by renal‐carcinoma cells treated with 5‐fluorouracil
Authors: Nishimura Naoki , Kanda Shigeru , Yogi Yasuo , Kawamura Masaya , Nakamura Mikio , Hyakutake Hiroyuki , Kanetake Hiroshi , Saito Yutaka
Published: February 2007

A study was made of the auto‐proliferative activity of human renal‐carcinoma cells in the supernatant from a carcinoma‐cell culture in serum‐free medium to which an anticancer agent had been added (5‐FU). The human renal‐cancer cells used in this study were of 3 strains: ACHN, VMRC‐RCW and NT. When each line was cultured in medium containing no 5‐FU, the supernatant showed almost no activity for stimulating DNA synthesis. However, when the line was cultured in the presence of 5‐FU, the supernatant showed autocrine growth‐promoting activity which strongly stimulated DNA synthesis of 3 renal‐cancer cell lines in a dose‐dependent manner. Activity could be detected in a 4‐ to 6‐kDa fraction by gel filtration. This fraction increased the DNA‐synthesis‐promoting activity of epidermal growth factor, transforming growth factor‐β, basic fibroblast growth factor and insulin, and was acid‐ and heat‐stable. It was also stable against pepsin and dithiothreitol. DNA synthesis in BALB/c 3T3, adult rat hepatocytes and rabbit renal tubular cells was not affected by this fraction which was thus considered not to affect non‐cancerous cells. Renal‐cell carcinoma responds poorly to anticancer agents, and the autocrine activity of the fraction may possibly be a factor accounting for this resistance. © 1992 Wiley‐Liss, Inc.
Human carcinoembryonic antigen cDNA expressed in rat carcinoma cells can function as target antigen for tumor localization of antibodies in nude rats and as rejection antigen in syngeneic rats
Authors: Pèlegrin André , Terskikh Alexei , Hayoz Daniel , Chalandon Yves , Olsson Nils‐Olivier , Folli Silvio , Buchegger Franz , Kromer Beril , Schwarz Klaus , Martin Monique , Martin Francois , Mach Jean‐Pierre
Published: February 2007

We have tried to develop a new model consisting of rats transplanted with syngeneic colon carcinoma PROb cells transfected with cDNA coding for the carcinoembryonic antigen (CEA), the human tumor marker most commonly used as target for MAbs. The antigenic density of the 4 CEA‐expressing clones selected for a precise characterization ranged from 5 × 104 to 1 × 106 CEA molecules per cell. In all clones the CEA was shown to be attached to the membrane by a phosphatidylinositol (PI) anchor. Using a panel of radiolabeled MAbs directed against the 5 major epitopes described on the CEA molecule, we showed that all these CEA epitopes were expressed by the 4 transfectants. Southern‐blot analysis showed that the entire CEA cDNA was present in the transfectants. Western‐blot analysis, however, showed that the size of the CEA expressed by the 4 transfectants was slightly smaller than that of CEA produced by 2 reference human colon‐carcinoma cell lines. Two clones, expressing 1 × 105 and 1 × 106 CEA molecules per cell, respectively, were grafted s.c. in nude mice and rats. Injection of radiolabeled anti‐CEA F(ab')2 fragments into these animals showed specific tumor localization with the highest percentages of injected doses for the transfectants expressing the highest CEA level. When grafted into immunocompetent syngeneic BDIX rats, the CEA‐expressing clones induced a strong antibody response against CEA and tumor rejections in a majority of the animals. Although the analysis of the immune response against the CEA‐cDNA‐transfected carcinoma cells is under investigation, the present results demonstrate that human CEA could function as a rejection antigen when transfected into rat carcinoma cells. © 1992 Wiley‐Liss, Inc.
Jumping translocations originate clonal rearrangements in SV40‐transformed human fibroblasts
Authors: Hoffschir F. , Ricoul M. , Lemieux N. , Estrade S. , Cassingena R. , Dutrillaux B.
Published: February 2007

A comparative study of chromosomal rearrangements occurring in 4 independent clones obtained from SV40‐transformed cornea and skin human fibroblasts was performed. Rearrangements principally affect some constitutive heterochromatin and, to a lesser degree, telomeric regions. This results in multiple exchanges between a limited number of chromosome structures, i.e., in jumping translocations. Such rearrangements occur even at early passages and some of them give rise to clonal rearrangements that accumulate at late passages. This process is responsible for progressive modification of the karyo‐types, principally characterized by deletion of a number of chromosome segments. Thus, clonal rearrangements are selected among many others not occurring at random. The selective pressure retaining clonal rearrangement seems to be similar for the 4 independent clones, since selection of the derivative chromosomes leads to the same imbalances, whatever the origin of the clone. This sequence of events recalls that of human solid tumors, since jumping rearrangements are generally observed in pre‐malignant conditions or in low‐grade malignancies, whereas clonal rearrangements leading to typical imbalances are detected in more advanced malignant tumors. © 1992 Wiley‐Liss, Inc.
Effect of MHC class‐I transfection on local tumor growth and metastasis in an H‐2‐negative clone derived from a chemically induced fibrosarcoma
Authors: Mialdea M. J. , Martin J. , Gaforio J. J. , Algarra I. , Pérez M. , Gonzalez A. , Ljunggren H. G. , Garrido F.
Published: February 2007

GR9 is a chemically induced fibrosarcoma composed of clones with different H‐2 class‐I expression. These clones differ with respect to local growth and spontaneous metastasis. The B9 clone (H‐2 negative) is highly tumorigenic (local growth) but of low metastatic potential (spontaneous metastasis assay). We have analyzed the effect that transfection of H‐2Dd and H‐2Kdgenes on this clone have upon local growth, lung colonization after i.v. injection and ability to form spontaneous metastases. The results showed that the effect on local growth of transfection of the Kd‐gene was stronger than that of the Dd gene. In addition, B9 co‐transfected with H‐2Kd and Dd genes showed the highest immunogenic properties in syngeneic BALB/c mice. Interestingly, the pSV2‐neo transfected clone gave almost the same result as that obtained with Dd transfection. Lung colonization after i.v. injection of the different clones (experimental metastasis), paralleled the results obtained for local growth: the number of lung nodules followed the cadence KdDd < Kd < Dd < pSV2. Survival of mice was always inversely correlated with local growth, e.g., all mice injected with 5 × 105 B9 H‐2KdDdtransfected cells survived. In contrast, no mice injected with the B9 control did. These differences were abrogated in irradiated and nude BALB/c mice. Finally, all transfected clones remained non‐metastatic in a spontaneous metastasis assay, behaving as the control, non‐transfected B9 cells. © 1992 Wiley‐Liss, Inc.
p53 over‐expression is an early event in the development of human squamous‐cell carcinoma of the larynx: Genetic and prognostic implications
Authors: Dolcetti Riccardo , Doglioni Claudio , Maestro Roberta , Gasparotto Daniela , Barzan Luigi , Pastore Antonio , Romanelli Massimiliano , Boiocchi Mauro
Published: February 2007

The tumor‐suppressor protein p53 is over‐expressed in a large fraction of squamous‐cell carcinomas of the larynx (LSCCs). p53 overexpression is dependent upon the synthesis of mutated versions of the protein and has been associated with the malignant progression of certain tumor types. In order to examine the prognostic value of p53 immunodetection in LSCCs, we performed a retrospective analysis on a selected series of tumors, using the PAb 1801 and CMI antibodies. No significant difference in the frequency of p53 over‐expression was observed between tumors from patients with early relapse (67%) and those who had been disease‐free for more than 5 years (84%). The lack of correlation of p53 immunoreactivity with clinical stage and differentiation grade of LSCCs, together with the coordinated expression of pS3 in primary tumors and the corresponding lymph‐node metastases, indicate that p53 over‐expression is probably unrelated to the biological aggres‐siveness of these tumors. In addition, the detection of p53 immunostaining in pre‐invasive areas as well as in preneoplastic lesions suggests that p53 abnormalities probably constitute a very early event in LSCC development. © 1992 Wiley‐Liss, Inc.
pS2 protein and steroid hormone receptors in invasive breast carcinomas
Authors: Koerner Frederick C. , Goldberg Diane E. , Edgerton Susan M. , Schwartz Laurent H.
Published: February 2007

We have used immunohistochemical and hormone‐binding techniques to determine the presence of estrogen receptors, estrogen‐receptor protein, progesterone receptors and pS2 protein in 97 invasive breast cancers. Our group of tumors displayed the same frequency of estrogen receptors and progesterone receptors as other comparable groups, but it contained more tumors containing pS2 protein. We also observed staining of morphologically normal cells in lobules adjacent to the tumors and in several fibroadenomas; both findings vary from some other reports. We attribute these variations to the use of a sensitive immunohistochemical method and choice of the lowest possible threshold to classify a tumor as pS2‐positive. If we used a higher threshold, then about 95% of the tumors containing pS2 protein also contained estrogen receptor protein. Our results add further weight to the assertion that tumors containing pS2 protein also display estrogen receptors. The data also provide theoretical and indirect support to the clinical prediction that tumors containing pS2 protein are more likely to respond to hormonal therapy and may have a more indolent course than tumors lacking the molecule. © 1992 Wiley‐Liss, Inc.
Progression of human cutaneous melanoma is associated with loss of expression of c‐kit proto‐oncogene receptor
Authors: Natali P. G. , Nicotra M. R. , Winkler A. B. , Cavaliere R. , Bigotti A. , Ullrich A.
Published: February 2007

Mutations at the white spotting (w) locus in mice have deleterious effects on germ cells, melanocytes and hematopoietic stem cells. The w locus encodes the c‐kit tyrosine‐kinase receptor whose ligand is the product of the SI locus. Using monoclonal antibodies (MAb(s) ) to the extracellular domain, we have evaluated the expression of c‐kit in normal and transformed melanocytes. This cell lineage synthetizes a receptor with a mw of 145 kDa. The gene product is expressed in epidermal melanocytes and in a fraction of nevocytic and blue nevi. In primary melanomas, loss of the receptor is observed in more invasive lesions. Only 30% of the metastatic lesions express detectable levels of the receptor. These findings demonstrate that the c‐kit product is down‐regulated in melanocytes following malignant transformation. The functional relevance of this modulation remains to be evaluated. © 1992 Wiley‐Liss, Inc.
Phenotypic overlaps between pleomorphic malignant T‐cell lymphomas and mixed‐cellularity Hodgkin's disease
Authors: Tosi Piero , Leoncini Lorenzo , del Vecchio Maria Teresa , Spina Donatella , Lorenzini Letizia , Barbini Paolo , Massai Maria Rita , Pileri Stefano , Sabattini Elena , Kraft Rainer , Cottier Hans
Published: February 2007

Histologically diagnosed, or in part questionable, malignant pleomorphic peripheral T‐cell lymphomas (pPTCLs, n = 16) and mixed‐cellularity Hodgkin's disease (MCHD, n = 12) were objectively compared by the use of combined immunohistochemistry on paraffin sections, test‐point analysis of tissue components, and semi‐automated nuclear morphometry on semi‐thin resin sections. Classical, qualitative histomorphological distinction of these sub‐types of lymphomas proved to be valid and is probably still the best method. Quantitative discriminant features, in order of decreasing significance, were: (i) expression by large atypical cells (LACs) of CD45RO, CD43 and CD45 in pPTCLs, and of CD30 and GD15 in MCHD; (ii) means and standard deviations (SDs) of LAC nuclear‐profile areas (greater in MCHD than in pPTCLs); (iii) expression of CD3 by LACs in pPTCLs; (iv) prominence of small lymphoid cells in MCHD; (v) higher percentage of medium‐sized lymphoid cells in pPTCLs; and (vi) higher SDs of nuclear‐profile circularity factor of small lymphoid cells in MCHD. The medians of the largest nucleolar profile areas in LACs per field did not differ in pPTCLs and MCHD, but dispersion of individual values towards higher levels was significantly greater in the latter. Stepwise discriminant analysis of test point and nucleometric variables that best distinguished pPTCLs from MCHD revealed considerable overlaps, and questionable cases tended to be intermediate between the two. In conclusion, our results confirm and expand the notion of intra‐group heterogeneity, with indistinct borders and the existence of intermediate phenotypes between these two taxonomic categories of malignant lymphomas. © 1992 Wiley‐Liss, Inc.
Expression of HER‐2/neu in renal‐cell carcinoma. Correlation with histologic subtypes and differentiation
Authors: Rotter Michael , Block Thomas , Busch Raymonde , Thanner Stefanie , Höfler Heinz
Published: February 2007

Twenty‐four renal‐cell carcinomas (RCC) and corresponding non‐neoplastic kidney tissue were examined for amplification and expression of the HER‐2/neu gene. Gene amplification was examined by slot‐blot analysis, mRNA expression by in situ hybridization and Northern blot analysis, and protein expression by immunohistochemistry. Northern‐blot analysis revealed lower expression of HER‐2/neu mRNA in clear‐cell (p < 0.001) and compact (p < 0.001) tumor subtypes, while chromophilic, chromophobic and tubulo‐papillary subtypes did not show significant differences in HER‐2/neu gene expression, as compared with non‐neoplastic kidney tissues. HER‐2/neu gene expression was not significantly associated with tumor stage. Low differentiation (G3) was associated with lower HER‐2/neu gene expression, but the number of G3 cases was too small for statistical analysis. HER‐2/neu gene amplification was not found in any of the tumors. The results of in situ hybridization and immunohistochemistry generally agreed with those of Northernblot analysis. We conclude that HER‐2/neu gene expression correlates with Thoenes' classification of RCC and may be inversely related to tumor differentiation; it is probably not involved in progression of RCC, in contrast to carcinomas of other locations (e.g. breast, ovary). © 1992 Wiley‐Liss, Inc.
Prevalence and distribution of human papillomavirus type‐16 DNA in pelvic lymph nodes of patients with cervical cancer and in women with no history of cervical abnormality
Authors: Rose Barbara R. , Thompson Carol H. , Chantrill Lorraine A. , Tattersall Martin H. N. , Cossart Yvonne E.
Published: February 2007

The polymerase chain reaction (PCR) was used to investigate the prevalence and distribution of human papillomavirus (HPV)‐16 DNA in paraffin sections of all pelvic lymph nodes removed from 14 patients with Stage Ib‐cervical cancer at the time of resection of their primary tumours. The results were compared with those obtained from 8 women with no known history of cervical abnormality. In all, 22 cervical biopsies and 401 lymph nodes (296 paraffin blocks) were examined. Nine of the 14 cervical cancer patients had primary tumours that were positive for HPV‐16 DNA: only 3 of these had lymph nodes with histological evidence of metastasis, and HPV 16 DNA was detected in each of the corresponding paraffin blocks. HPV 16 DNA was also detected in varying proportions (8%‐92%) of the histologically‐negative lymph nodes from these women. There was no correlation between the HPV DNA‐positive lymph nodes and their proximity to the primary tumour. HPV‐16 DNA was not identified in any of the lymph nodes from the 5 women whose cancers were not HPV‐16‐related, or in those of women with no evidence of cervical abnormality. This preliminary survey suggests that HPV DNA is frequently transported from HPV‐16‐related cervical tumours to regional lymph nodes. However, its practical significance will not be clear until sufficient time has elapsed for correlation of the results with the clinical outcome. © 1992 Wiley‐Liss, Inc.
Determination of placental ferritin (PLF)‐positive lymphocytes in women in early stages of breast cancer
Authors: Rosen Harald R. , Stierer Michael , Göttlicher Josef , Wolf Hermann , Weber Renate , Vogl Erich , Eibl Martha
Published: February 2007

Previous studies have proved that a certain acidic isoform of ferritin is specifically synthesized by the placenta and breast‐cancer tissue. In this context it has been further reported that the determination of this so‐called placental isoferritin (PLF) on the surface of a subset of peripheral lymphocytes is highly specific and sensitive for early stage breast cancer. By use of the monoclonal antibody CM‐H‐9 and flow cytometry, the levels of placental ferritin (PLF)‐positive cells were determined in 133 female patients undergoing surgical excision of a controversial or highly suspicious lesion of the breast detected by mammography. In addition, 61 healthy blood donors served as controls. Values of PLF‐positive cells in breast cancer patients differed significantly from those found in women with benign diseases and healthy controls (3.87% vs. 1.55% and 2.02, respectively; p < 0.00001). Analysis of prognostic factors in breast cancer patients (tumor size, lymph‐node status, menopausal status, estrogen receptor status, histologic grading and grading subfactors) revealed significantly higher levels of PLF‐positive cells in lymph‐node‐negative patients compared with node‐positive patients (p < 0.00001). Furthermore, levels of PLF‐positive cells s lowed a significantly negative correlation with tumor size and nuclear polymorphism. In 15 patients who underwent a guidewire‐directed surgical biopsy for a non‐palpable, mammographically suspect lesion, 4 cases of cancer correlated with high values of PLF‐positive lymphocytes while only I patient with a enign histologic result exhibited more than 4% positive cells. © 1992 Wiley‐Liss, Inc.
Matrigel promotes retinoblastoma cell growth in vitro and in vivo
Authors: Albini Adriana , Melchiori Antonella , Garofalo Angela , Noonan Douglas M. , Basolo Fulvio , Taraboletti Giulia , Chader Gerald J. , Giavazzi Raffaella
Published: February 2007

Cells derived from retinoblastomas grow slowly in vitro and only very rarely form tumors in nude mice. Matrigel, a mixture of components normally found in basement membranes, promotes the growth of Y‐79 and WERI‐Rbl retinoblastoma (Rb) cells when added to suspension cultures of the 2 Rb cell lines. It also substantially increases cell adhesion in vitro. Y‐79 cells, seeded into a Matrigel matrix, form round colonies over a 3‐week period similar to those of control, weakly metastatic murine melanoma cells. In vivo, s.c. co‐injection of Matrigel with either Y‐79 or WERI‐Rbl cells into nude mice promotes retino‐lastoma tumor formation. Transplantation of as few as 1,000 cells allows for xenografting under these conditions, while no tumors were observed in the absence of Matrigel, even at 10 × 106 cells/inoculum. The tumors produced have the expected morphology and express an mRNA for a highly specific retina/retinoblastoma marker protein, the interphotoreceptor retinoid‐binding protein. Thus, the xenografts obtained maintain the original morphological and molecular characteristics of the injected cells and represent a useful model for in vivo studies of retinoblastoma growth and treatment. © 1992 Wiley‐Liss, Inc.
Over‐expression of P‐glycoprotein and glutathione S‐transferase PI in MCF‐7 cells selected for vincristine resistance in vitro
Authors: Whelan Richard D. H. , Waring Clare J. , Wolf C. Roland , Hayes John D. , Hosking Louise K. , Hill Bridget T.
Published: February 2007

This study has provided evidence that exposure of the wild‐type MCF‐7 human breast carcinoma cell line to the mutagen ethyl methane sulphonate (EMS), followed by selection in vincristine (VCR), resulted in a stably‐resistant subline, designated VCREMS, which expressed an approximately 14‐fold level of resistance to VCR. This VCREMS subline showed cross‐resistance (3‐fold) to adriamycin (ADR) and to etoposide (3‐fold), but not to cisplatin. The addition of a non‐toxic concentration of verapamil (6.6 μM) significantly enhanced VCR cytotoxicity only in the resistant subline. This resistance was associated with over‐expression of P‐glycoprotein (Pgp), but without a concomitant increase in Pgp mRNA or gene amplification. In addition, activities of total glutathione S‐transferases (GST) and glutathione peroxidase were elevated in this resistant subline, with over‐expression of the GST‐pi isozyme and its associated mRNA being identified, without gene applification. This VCR‐selected resistant MCF‐7 cell line therefore provides another example of a breast carcinoma subline in which there is co‐ordinate over‐expression of both Pgp and GST‐pi, without attributing a causal relationship to either event, and extends the range of anti‐tumour drugs known to elicit modifications in glutathione metabolism. © 1992 Wiley‐Liss, Inc.
The human squamous cervical carcinoma cell line, HOG‐1, is responsive to steroid hormones
Authors: White J. O. , Jones R. N. , Croxtall J. D. , Gleeson R. P. , Krausz T. , Pervez S. , Jamil A. , Guida L. , Beesley J. E. , Soutter W. P.
Published: February 2007

Growth of the human squamous cervical carcinoma cell line, HOG‐1, was stimulated in response to oestradiol in serum‐containing and chemically defined medium. The oestradiol‐stimulated growth could be inhibited by 4‐OH tamoxifen, progesterone and medroxyprogesterone acetate; the last 2 compounds also inhibited basal cell growth in serum‐containing and chemically defined media. The data are consistent with the sensitivity of human squamous cervical cancer to sex‐steroid hormones and suggest that endocrine therapies may be of benefit in this disease. © 1992 Wiley‐Liss, Inc.
Two new human cholangiocarcinoma cell lines and their cytogenetics and responses to growth factors, hormones, cytokines or immunologic effector cells
Authors: Shimizu Yukihiro , Demetris Anthony J. , Gollin Susanne M. , Storto Patrick D. , Bedford H. Melanic , Altarac Silvio , Iwatsuki Shunzaburo , Herberman Ronald B. , Whiteside Theresa L.
Published: February 2007

Two new human cholangiocarcinoma (CC) cell lines (CC‐SW‐1 and CC‐LP‐1) were established and maintained in culture for 2 years. Histologically, both original liver tumors were adenocarcinomas, and the cell lines exhibited morphologic eatures of moderately differentiated adenocarcinoma. Immunohistochemistry showed that both cell lines were strongly positive for cytokeratin AEI but negative for carbohydrate tumor‐associated antigen, CA19–9. Ultrastructural analysis of both cell lines showed the presence of tight junctional complexes and focally formed microvilli. Both CC cell lines were tumorigenic in nude mice. Cytogenetic analysis showed that both cell lines expressed highly aneuploid karyotypes with numerous structural and numerical deviations. CC‐SW‐1 was hypodiploid with numerous chromosome losses and structural rearrangements, while CC‐LP‐1 was hyperdiploid and displayed multiple additional chromosomes. Doubling times for the CC‐SW‐1 and CC‐LP‐1 cell lines in the presence of 15% fetal bovine serum were 72 hr and 180 hr, respectively. Growth of the CC‐SW‐1 cell line was significantly stimulated in the presence of insulin, while that of the CC‐LP‐1 cell line was significantly augmented by epidermal growth factor (EGF). In contrast, dexamethasone strongly inhibited proliferation of both cell lines in a dose‐dependent manner. Among various recombinant cytokines examined for effects on growth or surface antigen expression on CC cell lines, only interleukin 1‐β (ILI‐β) strongly inhibited growth of the CC‐LP‐1 cell line, while interferons (IFNs) or tumor necrosis factor‐alpha (TNF‐α) were mildly inhibitory. Both tumor cell lines were resistant to natural killer (NK) cells but sensitive to lymphokine‐activated killer (LAK) cells. Preincubation of tumor cells with IFN‐γ, IFN‐α or TNF‐α significantly decreased the susceptibility of each tumor cell line to lysis by LAK cells, and the change in sensitivity did not correlate with the expression of HLA antigens or intercellular adhesion molecule‐1 (ICAM‐1) on the surface of tumor cells. These 2 CC cell lines are expected to provide valuable information about cell biology of human CC. © 1992 Wiley‐Liss, Inc.