bluuebunny/crossref_metadata_2025 huggingface dataset:
Modulation of MDR‐1 expression by a H‐ras oncogene in a human colon carcinoma cell line
Authors: Kramer Robert , Arceci Robert , Weber Thomas K. , Morse Barry , Simpson Heather , Steele Glenn D. , Summerhayes Ian C.
Published: February 2007

In an established colon differentiation model, introduction of a c‐H‐ras‐1 oncogene into a poorly differentiated human colon carcinoma cell line (Clone A) results in changes associated with the acquisition of a more differentiated phenotype. Down‐regulation of mdr‐l mRNA was shown to accompany ras‐related differentiation events resulting in decreased Pgp synthesis and a significant reduction in membrane Pgp as detected by immunoprecipitation, Western‐blot and FACS analysis. Consistent with these observations was a reduction in Pgp‐mediated drug resistance associated with Clone‐A ras transfectants, with no alteration in drug sensitivity being observed with non‐MDR drugs in these cells. An alternative differentiation model involves exposure of Clone‐A cells to sodium butyrate. Under these conditions, differentiation‐related changes resulted in up‐regulation of mdr‐1 mRNA and Pgp synthesis, although no alteration in drug sensitivity was recorded. In agreement with this observation, the levels of membrane‐associated Pgp remained unchanged throughout the period of exposure to sodium butyrate. This study shows that modulation of Pgp expression in colon differentiation is dependent upon the differentiation induction agent used.
Dimethyl sulfoxide (DMSO) increases expression of sialyl lewis X antigen and enhances adhesion of human gastric carcinoma (NUGC4) cells to activated endothelial cells
Authors: Maehara Masanori , Isobe Yoshiaki , Nakagawara Gizo , Yagita Masato , Hoshino Takashi
Published: February 2007

Dimethyl sulfoxide (DMSO) exerts a number of biological effects including the promotion of cell differentiation in cultured cells. In this study, we examined the effect of DMSO on the adhesion of tumor cells to endothelial cells. In vitro treatment of human gastric adenocarcinoma (NUGC4) cells with DMSO resulted in increased adhesion to interleukin‐1 (IL‐l)‐activated human endothelial cells compared with DMSO‐untreated NUGC4 cells. In flow cytometry, treating NUGC4 cells with DMSO enhanced the expression of sialyl Lewis x (sialyl Lex) and sialyl dimeric Lex antigens on their surface. Also, the binding of Limulus polyphemus agglutinin (LPA), which specifically binds to cell‐surface sialic acids, was increased by DMSO. The adhesion of DMSO‐treated NUGC4 cells to activated endothelial cells was blocked by neuraminidase pre‐treatment of tumor cells or by antibody against either endothelial leukocyte adhesion molecule‐1 (ELAM‐I) or sialyl Lex. Thus, it is suggested that enhanced adhesion following DMSO treatment is mediated by the interaction of sialyl Lex expressed on NUGC4 cells with ELAM‐I of endothelial cells. The modulation of sialyl Lex antigen by DMSO provides a useful system for studying the regulatory mechanism of Lewis‐related carbohydrate antigens and also for understanding the metastatic properties of cancer cells.
Anomalous expression of P‐glycoprotein in highly drug‐resistant human KB cells
Authors: Dolci Elizabeth D. , Abramson Robin , Xuan Yongzhi , Siegfried Jennifer , Yuenger Kristin A. , Yassa David S. , Tritton Thomas R.
Published: February 2007

KB‐AI and KB‐A10 are 2 multi‐drug‐resistant cell lines which are 100‐ and 1,000‐fold resistant to Adriamycin, respectively. We have examined the expression of P‐glycoprotein at the molecular and cellular levels in these human carcinoma cells. Both MDR cell lines, when compared to the parental KB‐3–1, show characteristic increases in mdr l gene copy number, an increase in mdr l mRNA expression, a corresponding increase in transcription rate and a consequent over‐expression of P‐glycoprotein. However, the more highly resistant KB‐A 10 cells have a lower gene copy number, express less mdr l mRNA and contain less P‐glycoprotein than the AI cell line. To determine whether higher levels of cellular resistance were attributable to enhanced efficacy of P‐glycoprotein or to other cellular regulatory mechanisms, we examined other major cellular properties known to be associated with the mdr phenotype. Both the KB‐AI and KB‐A 10 lines exhibit similar increases in protein kinase C activity as compared to the drug‐sensitive parent. In addition, neither glutathione‐S‐transferase nor topoisomerase II activities account for enhanced resistance of the KB‐A 10 cells. The above observations are contrary to the premise that the level of drug resistance is necessarily proportional to expression of P‐glycoprotein or to other common factors thought to participate in drug insensitivity; consequently, new mechanisms of resistance must be in operation in these cells.
A study of cross‐resistance pattern and expression of molecular markers of multidrug resistance in a human small‐cell lung‐cancer cell line selected with doxorubicin
Authors: Supino Rosanna , Binaschi Monica , Capranico Giovanni , Gambetta Romolo A. , Sala Elena , Zunino Franco , Prosperi Ennio
Published: February 2007

A doxorubicin‐resistant variant of the human small‐cell lung‐cancer cell line N592 was selected by in vitro continuous exposure to increasing drug concentrations. The aim of this study was to examine the cross‐resistance pattern, cellular pharmacokinetics of doxorubicin and expression of molecular factors of resistance. The sub‐line N592/DX exhibited a multidrug‐resistance phenotype, which was somewhat atypical, since it included cisplatin. Development of doxorubicin resistance could not be attributed to differential doxorubicin uptake or retention. Verapamil partially reverted doxorubicin resistance without affecting cellular pharmacokinetics. These findings are consistent with undetectable levels of mdr‐1 ‐gene expression in these cells. A molecular analysis of other putative mechanisms of multidrug resistance indicated no alterations in GSH levels or GSH‐related enzymes, but a marginal reduction of topoisomerase II α expression in the resistant sub‐line. This reduction, which was associated with an increase in topoisomerase I, does not explain the high degree of resistance. This study supports the view that alternative, unidentified mechanisms, which may be of clinical relevance, must be involved in the development of multidrug resistance of small‐cell lung cancer.
Somatostatin receptor‐positive primary breast tumors: Genetic, patient and tumor characteristics
Authors: Bootsma A. H. , Lamberts S. W. J. , Van Eijck C. , Schouten K. K. , De Klein A. , Reubi J. C. , Waser B. , Foekens J. A. , Van Pel R. , Zwarthoff E. C.
Published: February 2007

In a series of 87 primary breast tumors, somatostatin receptor (SSR) expression was detected by in vitro autoradiography in 58 tumors. In 41 tumors the SSR expression was homogeneous and in 17 it was heterogeneous. Although the tumors were not selected by the investigators upon entry in the study, examination of the tumor and patient characteristics showed that a pre‐selection had taken place for small tumors. Eighty percent of the tumors were classified as stage pTI or pT2 tumors. This small tumor size and the large size of the tumor sections used for autoradiography can explain the high incidence of somatostatin expression in our series. Forty‐three of these tumors, 30 SSR‐positive and 13 SSR‐negative, were tested for morphological and (immuno)histochemical markers of neuroendocrine differentiation. Three SSR‐positive tumors were also positive for 2 or more other markers of neuroendocrine differentiation, suggesting that neuroendocrine breast tumors and SSR‐positive breast tumors are overlapping, but independent, subgroups of tumors. To test whether specific genetic alterations are associated with SSR‐positive or SSR‐negative breast tumors, we examined in a selected series of 47 SSR‐positive and 32 SSR‐negative breast tumors a number of known genetic markers by Southern blotting. Deletions or rearrangements of the retinoblastoma (RB) tumor‐suppressor gene were observed in 5 SSR‐positive and 5 SSR‐negative tumors. In 4 SSR‐positive and also in 4 SSR‐negative tumors an amplification of the neu oncogene was observed. Amplifications of the int‐2 oncogene were found in 2 SSR‐positive and I SSR‐negative breast tumor. In one SSR‐positive tumor an amplification of the c‐myc oncogene was observed and in another SSR‐positive tumor a rearrangement of the L‐myc oncogene was found.
p53 Mutations in sporadic adrenocortical tumors
Authors: Ohgaki H. , Kleihues P. , Heitz P. U.
Published: February 2007

Non‐familial human adrenocortical adenomas and carcinomas were screened for mutations in exons 5–8 of the p53 tumor suppressor gene by single‐strand‐conformation‐polymorphism (SSCp) analysis, followed by direct sequencing of PCR‐amplified DNA. point mutations in codons 12, 13 and 61 in H‐ras, K‐ras and N‐ras proto‐oncogenes were similarly assessed by direct DNA sequencing. Three out of 15 primary adrenocortical carcinomas (20%) contained a mis‐sense point mutation in the conserved regions (exons 5 and 8) of the p53 gene. Mutations were located in codon 157 (GTC → TTC; Val → phe), codon 163 (TAC → AAC; Tyr → Asn), and codon 273 (CGT → TGT; Arg → Cys). The mutation in codon 157 was detected in the primary tumor as well as in brain and lymph‐node metastases. Among 18 adrenocortical adenomas, there was only a single non‐miscoding mutation in codon 295 (CCT → CCC; pro → pro). These data suggest that mutational inactivation of the p53 gene occurs in a minority (20%) of sporadic adrenocortical carcinomas and that these mutations constitute a late event in the multi‐step process of malignant transformation. No ras mutations were detected in any of these tumors, suggesting that these genes are not involved in the development of tumors originating from the adrenal cortex.
Biodistribution study of 99mTc‐labeled LDL in B16‐melanoma‐bearing mice. Visualization of a preferential uptake by the tumor
Authors: Ponty Emmanuelle , Carton Michel , Soula Georges , Favre Gillcs , Benaniba Rachid , Boneu Andrée , Lucot Henriette
Published: February 2007

Since there is strong evidence of a preferential LDL accumulation in tumor cells, LDL might be of interest for tumor imaging. We have tested the ability of 99mTc‐LDL in tumor imaging with B16‐melanoma‐bearing mice as a model for further applications in human studies. The LDL fixation rate was higher with 99mTc‐labeled LDL than with 125I labeled LDL. Since technetium‐99m remains trapped in the cells, 99mTc‐LDL is a well‐adapted radioligand because of information given by this radiotracer on the receptor metabolism. We observed that, at early growth stages, the tumor took up the LDL at a maximal rate, suggesting differences in cholesterol metabolism as a function of tumor growth. Accumulation of label in the tumor area was perfectly observable in tumor‐bearing mice on scintigraphic images. Computerized quantification of the regions of interest (as well as biodistribution studies including killing of the animals) showed a 1.81 ‐fold increase in uptake by the tumor as compared to the liver and a 28‐fold increase as compared with corresponding normal tissue (muscle of the left leg) at day 8 of tumor growth. These data give strong support to the value of this non‐invasive method in visualizing and quantifying the tissue LDL uptake in vivo, including the precise information provided by nuclear scintigraphy on the distribution of the radiolabeled LDL in the different tissues. 99mTc‐LDL could be an efficient tool for further diagnostic or therapeutic exploration in cancer patients.
CD30‐specific AB1‐AB2‐AB3 internal image antibody network: Potential use as anti‐idiotype vaccine against Hodgkin's lymphoma
Authors: Pohl C. , Renner C. , Schwonzen M. , Schobert I. , Liebenberg V. , Wolf J. , Diehl V. , Jung W. , Pfreundschuh M.
Published: February 2007

The tumor‐associated CD30 antigen is presently under study as a target for active specific immunotherapy of Hodgkin's lymphoma with anti‐idiotypic antibodies. Internal image antibodies (Ab2(i) 9G10 and 14G9 against the CD30‐specific antibody HRS‐4 (Abl) have been described, which induce a CD30‐specific T‐ and B‐cell response in BALB/c mice and New Zealand white rabbits. In extension of this work, murine monoclonal anti‐idiotypic Ab2β 9G10, mimicking structures of the nominal CD30 antigen, was used to generate monoclonal Ab3 in mice and polyclonal Ab3 in rabbits with specificity for CD30. The Ab2p 9G10‐specific murine monoclonal Ab3 4A4 bound specifically to the 120‐kDa band of CD30 present on Hodgkin cell lines and Hodgkin tumor tissue, and effectively inhibited binding of Ab1 HRS‐4 to Ab2 9G10 as well as to CD30+ cells. Monoclonal Ab3 4A4 was cytotoxic for CD30+ cell lines in vitro and effectively prevented the s.c. growth of L540 cell tumors after passive i.v. administration in a SCID mouse tumor model. While this cytotoxic effect of the IgM subclass monoclonal Ab3 4A4 was due to complement activation, the murine monoclonal Abl HRS‐4 and a polyclonal Ab3 preparation of 1gG‐subclass from New Zealand white rabbits were cytotoxic by an antibody‐dependent cell‐mediated mechanism in vitro. In conclusion, Ab2p 9G10 is able to induce a CD30‐specific cytotoxic 1gG and 1gM response. Cytotoxicity was shown to be mediated by complement activation and antibody‐dependent cell‐mediated cytotoxicity in vitro and in vivo and across species barriers. Thus, the CD30‐like Ab2β 9G10 may hold promise for effective active specific immunotherapy of human Hodgkin's lymphoma.
Intracellular glutathione heterogeneity in L1210 murine leukemia sublines made resistant to dna‐interacting anti‐neoplastic agents
Authors: Tagliabue Giovanna , Ubezio Paolo , Balconi Giovanna , Mascellani Enrico , D'Incalci Maurizio , Pifferi Antonio , Geroni Cristina
Published: February 2007

Intracellular glutathione (GSH) content was measured by flow cytometry using monochlorobimane (mBCI) and by the enzymatic assay in a set of 6 sublines of murine L1210 leukemia cells made resistant to DNA‐interacting agents having distinct mechanisms of action: L‐phenylalanine mustard (L‐PAM), 1,3‐bis(2‐chloroethyl)‐1‐nitrosourea (BCNU), cisplatin (DDP), N‐de‐formyl‐N‐(4‐N, N‐bis(2‐chloroethylamino) benzoyl) distamycin A (FCE 24517), doxorubicin (DX) and 3′ ‐deamino‐3′ (2‐methoxy‐4‐morpholinyl)‐doxorubicin (FCE 23762). A significant correlation was demonstrated between the mean intracellular mBCI fluorescence values measured by flow cytometry and levels of GSH measured by the classical enzymatic assay, despite the possible influence of glutathione‐S‐transferases and of other thiols on the mBCI fluorescence. Although less specific, the flow cytometric method is more informative than the enzymatic assay, allowing detection of fluorescence distributions, which we proved to be characteristic of each subline. In order to assess a procedure enabling a quantitative analysis to be made of intercellular GSH heterogeneity, we propose the use of appropriate thresholds and parameters of the mBCI flow cytometric distribution. By use of this analysis procedure, distinct types of alterations, with respect to the heterogeneity distribution of the parental L1210 cell line, have been evidenced in resistant cells. A uniform increase in mBCI fluorescence was observed among cells of the sublines resistant to L‐PAM and FCE‐24517. The mean mBCI fluorescence increase in sublines resistant to DX and DDp was due to a higher number of cells with fairly high mBCI fluorescence, but still within the range spanned by the parental cell line. A less heterogeneous mBCI fluorescence distribution was found in the L1210 subline resistant to FCE 23762, which was, however, similar to a cloned sensitive line. Though GSH was linked to the principal cause of drug resistance only in the L‐PAM‐resistant cell line, alterations in heterogeneity, as detected by mBCI fluorescence distributions, were found in 5 out of 6 resistant lines.
Inhibition of glycolysis and mitochondrial respiration of ehrlich ascites carcinoma cells by methylglyoxal
Authors: Halder Jyotsnabaran , Ray Manju , Ray Subhankar
Published: February 2007

The effect of methylglyoxal (MG) on the aerobic glycolysis of Ehrlich ascites carcinoma (EAC) cells has been tested. Methylglyoxal inhibited glucose utilization and glucose 6‐phosphate (G6P) and L‐lactate formation in whole EAC cells. Methylglyoxal strongly inactivated glyceraldehyde 3‐phosphate dehydrogenase (GA3PD) of the malignant cells, whereas MG has little inactivating effect on this enzyme from several normal sources. Methylglyoxal also inactivated only the participate hexominase of the EAC cells, but this inactivation was less pronounced than the effect on GA3PD. Methylglyoxal has little inactivating effect on glucose 6‐phosphate dehydrogenase (G6PD), and no effect on L‐lactate dehydrogenase (LDH) of the malignant cells. Glucose‐dependent L‐lactic acid formation of EAC‐cell‐free homogenate was strongly inhibited by MG, but when GA3PD of normal cells was added to this homogenate, significant lactate formation was observed even in the presence of MG. Methylglyoxal also inhibited the respiration of EAC‐cell mitochondria. Respiration of mitochondria isolated from liver and kidney of normal mice, however, remained unaffected. As a consequence of the inhibition of glycolysis and mitochondrial respiration, the ATp level of the EAC cells was drastically reduced. Studies reported herein strongly suggest that the tumoricidal effect of MG is mediated at least in part through the inhibition of mitochondrial respiration and inactivation of GA3PD, and this enzyme may play an important role in the high glycolytic capacity of the malignant cells.
Transformation of mouse fibroblasts with the oncogenes H‐ras or trk is associated with pronounced changes in drug sensitivity and metabolism
Authors: Peters G. J. , Wets M. , Keepers Y. P. A. M. , Oskam R. , Van Ark‐Otte J. , Noordhuis P. , Smid K. , Pinedo H. M.
Published: February 2007

Malignant activation of oncogenes ras or trk is implicated in a number of solid tumors and leukemias. We determined the chemosensitivity profile of wild‐type mouse NIH‐3T3 fibro‐blasts, and that of NIH‐3T3 lines transformed by the H‐ras (S2‐721) and trk (106–632) oncogenes, against 11 different drugs from various classes. Differences in sensitivity were related to drug accumulation and metabolism. Both ras‐ and trk‐trans‐formed cell lines were less sensitive to cisplatin (CDDp) and doxorubicin (DXR) than the wild type. NIH‐3T3 transformants expressing H‐ras were less sensitive than those expressing trk or the wild type to the indoloquinone EO9, methotrexate and arabino‐furanosylcytosine. No clear difference in sensitivity was observed for vincristine, Vp‐16, or the new cytidine analog 2′, 2′ ‐difluoro‐deoxycytidine. In both ras‐ and trk‐transformed cell lines sensitivity to 5FU was increased moderately, but sensitivity to 5′ deoxy‐5‐fluorouridine (5′dFUR) was increased markedly. Only the trk‐transformed line NIH‐3T3 was more sensitive to 2′ deoxy‐5‐fluorouridine. Expression of P‐glycoprotein was not different between the 3 cell lines but DXR accumulation in both mutants was decreased, indicating a non‐P‐glycoprotein‐associated difference in sensitivity. Conversion of 5′ dFUR to 5FU (catalyzed by pyrimidine nucleoside phosphorylases) was 5–10 times higher in both mutants than in the wild type. The activity of the phosphoribosyl‐transferase (direct conversion of 5FU to FUMP) was comparable, but the rate of conversion of 5FU to fluorouridine (FUR) was lower in the wild type, as well as that of 5FU to FUMP via FUR. In contrast, the activity of thymidylate synthase, the target enzyme for fluoropyrimidines, was higher in the wild‐type cells. The concentrations of both purine and pyrimidine nucleotides were lower in cells expressing trk. In conclusion, transformation of cells with the H‐ras or trk oncogenes can markedly influence sensitivity to several drugs and affect normal metabolism and that of several anti‐cancer agents.
The effect of ion channel blockers, immunosuppressive agents, and other drugs on the activity of the multi‐drug transporter
Authors: Weaver James L. , Pine P. Scott , Aszalos Adorjan , Szabo Gabor , Gottesman Michael M. , Goldenberg Sarah
Published: February 2007

The MDRI protein is an energy‐dependent transport protein responsible for the multi‐drug resistance seen in many tumors. A variety of drugs have been shown to inhibit the function of this pump, including compounds known to block various ion channels. The mouse lymphoma cell line L5178Y has been transduced with the human mdr 1 gene. Using this cell line, we have tested a number of compounds to determine whether there is a correlation between the ability to block a specific type of ion channel, or shift membrane potential, and the ability to act as an MDR‐reversing agent using the fluorescent substrates Rhodamine 123 and daunorubicin as test compounds. Our results show no apparent correlation between the ability to block a specific ion channel and reversal of MDR transport ability. We have found active MDR inhibitors in compounds that affect K+, Na+, Ca+ +, H+, but not Cl− channels. Our data suggest that Cl−channel activity may be distinct from MDR activity. Several immunosuppressive compounds and analogs were also tested and found to be active reversing agents. Measurements suggest a significant difference in resting membrane potential between the L5178YvMDR line and the L5178Y parental cell line used in these experiments. No correlation was found between the ability of drugs to alter membrane potential and to inhibit MDR transport activity. Our results suggest that MDR transport function may be independent of the physiological movement of ions and show that a wide variety of compounds can inhibit MDR transport.
Monoclonal antibody directed against colon cancer mucin has high specificity for malignancy
Authors: Siddiki Badcr , Ho Jenny J. L. , Huang Jian , Byrd James C. , Lau Eva , Yuan Mei , Kim Young S.
Published: February 2007

Exposure of core carbohydrate structures such as NeuAc alpha2–6GalNAc‐Ser/Thr (sialosyl‐Tn) in mucus glycoproteins is often associated with malignant transformation in a number of different tissues. Reagents that specifically identify such structures would be useful in the diagnosis of cancer. Monoclonal antibody JT10e has been produced against mucins from xeno‐grafts of LS174T colon cancer cells but also reacts with mucins of pancreatic cancer xenografts. The following data suggest that JTlOe reacts with sialosyl‐Tn: (I) reactivity with bovine and ovine submaxillary mucins; (2) reactivity sensitive to neuramini‐dase or mild acid; (3) inhibition of reactivity by NeuAc and NeuAc alpha2–6 lactose; and (4) lack of reactivity with other glycoproteins with related carbohydrate structures but no sialosyl‐Tn. JTlOe is distinguishable from 2 other antibodies which react with sialosyl‐Tn, B72.3 and TKH2 because JT10e: (a) did not react with normal gastric tissue while B72.3 and TKH2 did; (b) was only partially inhibited by TKH2 and not at all by B72.3; (c) did not react with Tn antigen while B72.3 did; and (d) bound more strongly to pancreatic cancer mucins than did either B72.3 or TKH2. JT10e reacted with a high percentage of malignant colonic, pancreatic, gastric and mammary tissues but not with the corresponding normal tissues. A high percentage of patients with colonic, pancreatic, gastric, mammary and lung cancers had elevated blood levels of JT10e antigen. A number of colonic cancer patients had elevated JT10e antigen levels without corresponding elevations in CA19–9 levels. These results suggest that JT10e antibody could be used in conjunction with other mucin markers to improve the identification of malignancy in colon, and to study the structure of oligosaccha‐rides in cancer mucins.
Sensitized T lymphocytes render DBA/2 beige mice responsive to IFN α/β therapy of friend erythroleukemia visceral metastases
Authors: Kaido Thomas , Gresser Ion , Maury Chantal , Vignaux Françoise , Maunoury Maric‐Thérèse , Belardelli Filippo
Published: February 2007

Interferon α/β (IFN α/β) is highly effective in inhibiting the development of Friend erythroleukemia cell (FLC) visceral metastases in DBA/2 mice injected intravenously (i.v.) with FLC, but does not protect FLC‐injected DBA/2 beige (bg/bg) mice. Use of IFN α/β‐resistant FLC indicated that IFN was acting through host mechanisms in DBA/2 mice and thus pointed to a defect in some host mechanism in bg/bg mice essential for IFN's anti‐metastatic action. We undertook experiments to restore in bg/bg mice the marked anti‐FLC meta‐static effect of IFN a/p observed in DBA/2 and +/bg mice. Adoptive transfer of spleen cells from normal syngeneic mice to IFN‐treated bg/bg mice was ineffective, but the transfer of splenic T lymphocytes from FLC‐immunized DBA/2 or +/bg mice markedly increased the survival time of FLC‐injected bg/bg mice provided that these mice were also treated with IFN a/p. Neither treatment alone resulted in an increase in survival time. As few as 1 × 107 immune spleen cells were effective in IFN‐treated FLC‐injected bg/bg mice. The T‐cell immune response to FLC of bg/bg mice was diminished compared with that of +/bg mice. Likewise, only combination therapy of immune spleen cells and IFN α/β resulted in an increased survival time of ESb‐lymphoma‐injected bg/bg mice. Our results indicate the essential participation both of T‐cell‐mediated immune mechanisms and of IFN α/βin the inhibition of FLC visceral metastases.
Differential expression and release of LFA‐3 and ICAM‐1 in human melanoma cell lines
Authors: Scheibenbogen Carmen , Keilholz Ulrich , Hunstein Werner , Meuer Stefan , Dengler Thomas , Tilgen Wolfgang
Published: February 2007

We examined 10 different melanoma cell lines for cellular expression and release of I CAM‐1 (CD54) and LFA‐3 (CD58) and the influence of cytokines, including IFNa, IFNγ and TNFa. Cellular ICAM‐I expression and density varies considerably between the melanoma cell lines. While IFNα has no effect on cellular ICAM‐I (cICAM‐l) expression, IFNγ and to a lesser extent TNFα can effectively up‐regulate cICAM‐l. Soluble I CAM‐1 (sICAM‐1) is detected in the supernatants of all lines tested, release of sICAM‐1 correlates with cellular expression. LFA‐3 does not much differ in its expression level on melanoma lines, and cytokines have little or no effect on its expression. Soluble LFA‐3 is released by only 6 out of 10 lines. Its release can effectively be inhibited by IFNγ in all lines and by TNFa in one, while IFNa has no effect. These data show that expression and release of LFA‐3 and ICAM‐I differ between melanoma cell lines. This may be of importance for the interaction of melanoma cells with immune effector cells in vivo.
Disialoganglioside GD2 anti‐idiotypic monoclonal antibodies
Authors: Cheung Nai‐Kong V. , Canete Adela , Cheung Irene Y. , Ye Jian‐Nan , Liu Chongyuan
Published: February 2007

Disialoganglioside GD2 is widely expressed among neuroblas‐tomas, melanomas, small‐cell lung carcinoma, sarcomas and brain tumors. Immunity directed against this antigen may have anti‐tumor utility. Since GD2 is poorly immunogenic, anti‐idiotypic antibodies may serve as alternative tumor vaccines. Monoclonal antibody 3F8, a murine lgG3 specific for GD2, has shown excellent tumor‐targeting ability in vitro and in vivo. LOU/CN rats were immunized with 3F8 and their spleens were used in somatic‐cell hybridization, using Sp2/0, p3 and Y3 as fusion partners. Six anti‐idiotypic (anti‐id) MAbs (C2D8, ldio‐2, AIG4, C2H7, C4E4, A2A6) were selected based on their reactivity with 3F8 and non‐reactivity with murine lgG3 myelomas. Specificity of each anti‐id was demonstrated by using various ELISA: (i) lack of direct binding to solid phase myelomas and serum proteins; (ii) inability of other myelomas to inhibit anti‐id binding to 3F8; (iii) absence of cross‐reactivity of other myelomas to solid‐phase anti‐id; (iv) lack of inhibition by anti‐id of binding of other ganglioside antibodies to their antigens. Antigen specificity was further examined by inhibition of binding of 3F8 to GD2 on immuno‐thin‐layer chromatography, and by inhibition of 3F8 immunostaining of neuroblastoma cell lines. These 6 antibodies were demonstrated to be distinct, in view of their cross‐reactivity, fusion partners and relative strength of binding to 3F8. Anti‐GD2 antibodies were induced after immunization with these anti‐id antibodies in C57BI/6 mice. These rat anti‐3F8‐idiotypic antibodies with exquisite specificity for anti‐GD2 antibodies may be useful in vaccine construction.
Dualistic effects of cis‐diammine‐dichloro‐platinum on the anti‐tumor efficacy of subsequently applied recombinant interleukin‐2 therapy: A tumor‐dependent phenomenon
Authors: Bernsen Moniquc R. , Heintz A. Peter M. , Van Barungen Harrold J. J. , Van Der Velden Alike W. , Dullens Hub F. J. , Otter Willem Den
Published: February 2007

The efficacy with which disseminated SL2 and P815 tumors, in syngeneic DBA/2 mice, can be eradicated with low‐dose recombinant interleukin‐2 (rlL‐2) therapy is about equal. Treatment (i.p.) of DBA/2 mice, injected i.p. with SL2 or P8I5 cells on day 0, with rlL‐2 (Proleukin) on days 10 to 14 results in a cure rate of 50 to 60% in each case. The in vitro sensitivity of SL2 and P8I5 cells to cis‐diammine‐dichloro‐platinum [II] (cisplatin) is also comparable, although P8I5 appears to be slightly more sensitive. In vivo, however, therapy with cisplatin is far less effective against P8I5 than against SL2. In the DBA/2‐SL2 model, at all doses tested, combination therapy with cisplatin (administered on day 2) and rlL‐2 (administered on days 10–14) resulted in anti‐tumor efficacy greater than that of either drug separately. In contrast, in the DBA/2‐P8I5 model, cisplatin decreased the anti‐tumor efficacy of subsequently applied rlL‐2 therapy. As the only difference between the 2 tumor models is the tumor itself, the success of combination therapy with cisplatin and rlL‐2 was dependent on tumor characteristics. We suggest that in these 2 tumor models, neither the sensitivity of these tumors to cisplatin nor their growth and dissemination patterns were responsible for the contrasting results of combination therapy in these models. Instead, tumor‐dependent immune‐modulating effects of cisplatin may be the cause of these effects. These immune‐modulating effects may comprise (a) effects of cisplatin on the tumor cells, resulting in changes in their susceptibility to immune effector cells, or changes in their immunogenicity; (b) activating or suppressive effects of cisplatin on immune effector cells; or (c) a combination of these effects. These effects could then either synergize or antagonize with the immune activating properties of rlL‐2.
Increased risk of nasopharyngeal carcinoma among males of french origin born in maghreb (north africa)
Authors: Jeannel D. , Ghnassia M. , Hubert A. , Sancho‐Garnier H. , Eschwège F. , Crognier E. , De‐Thé G.
Published: February 2007

A study on the incidence of nasopharyngeal carcinoma (NPC) has been performed in the Provence—Alpes—Cote d'Azur (PACA) region, South of France, where Maghrebian migrants represent 10% of the population. During the years 1986–1990, 76 cases of NPC were diagnosed, of which 44 were French by birth, 25 were Maghrebian migrants and 7 were other migrants. We found that 36% of the French patients were either born in endemic areas or had lived there for more than 15 years. Standardized annual incidence rates of NPC among Maghrebian migrants remained close to those observed in North Africa, while, standardized annual incidence rates for French by birth in the PACA region were similar to those observed in cancer registries in other regions of France and countries of low incidence. However, for males of French origin born in Maghrebian countries, the incidence of NPC (all cases being of undiffer‐entiated type) was 5.7 times higher than that of males of French origin born in France (p = 0.00001). These results favor a critical role of environmental factors in the risk of NPC.
Allelic loss in ovarian cancer
Authors: Yang‐Feng Teresa L. , Han Hong , Chen Kuang‐Chao , Li Shi‐Bo , Claus Elizabeth B. , Carcangiu Maria L. , Chambers Setsuko K. , Chambers Joseph T. , Schwartz Peter E.
Published: February 2007

Loss of heterozygosity (LOH) was examined at 86 loci distributed on every chromosomal arm in 50 human ovarian tumors. Frequent allele losses were observed on chromosomes 13q (42%), 17p (42%), 17q (45%), and Xp (41%). Deletion mapping on chromosome 17 revealed a candidate gene on the long arm distal to D17S41/S74 for ovarian cancer which is distant from the locus for early onset breast cancer. LOH on chromosome 17q was found to be concordant with LOH on chromosomes 3p, 13q, 17p and Xp suggesting that it may be an early event in neoplastic development. These findings indicate that multiple tumor‐suppressor genes for ovarian cancer possibly exist on chromosomes 13q, 17, and/or Xp and provide the basis for the identification of candidate gene(s) associated with ovarian cancer. The chromosomal mechanisms resulting in allele losses in ovarian cancer include deletion, deletion/duplication, mitotic recombination and monosomy, in concordance with the developed genetic model.
Estimates of the worldwide incidence of eighteen major cancers in 1985
Authors: Parkin D. M. , Pisani P. , Ferlay J.
Published: February 2007

The annual incidence rates (crude and age‐standardized) and numbers of new cases of 18 different cancers have been estimated for the year 1985 in 24 areas of the world. The total number of new cancer cases (excluding non‐melanoma skin cancer) was 7.6 million, 52% of which occur in developing countries. The most common cancer in the world today is lung cancer, accounting for 17.6% of cancers of men worldwide, and 22% of cancers in men in the developed countries. Stomach cancer is now second in frequency (it was slightly more common than lung cancer in 1980) and breast cancer—by far the most important cancer of women (19.1% of the total)—is third. There are very large differences in the relative importance of the different cancers by world area. The major cancers of developed countries (other than the 3 already named) are cancers of the colon‐rectum and prostate, and, in developing countries, cancers of the cervix uteri, mouth and pharynx, liver and oesophagus. The implications of these patterns for cancer control, and specifically prevention, are discussed. Tobacco smoking and chewing are almost certainly the major prevent able causes of cancer today.
Expression of MMP‐7(pump‐1) mRNA in human colorectal cancers
Authors: Yoshimoto Mitsuru , Itoh Fumio , Yamamoto Hiroyuki , Hinoda Yuji , Imai Kohzoh , Yachi Akira
Published: February 2007

The expression of MMP‐7 (pump‐1) gene was examined in 10 cases of colorectal cancer by utilizing RT‐PCR. In 9 out of 10 cases, MMP‐7 mRNA was detected in cancerous tissue, whereas none was detected in adjacent normal colon tissue. However, this message was detected in only I out of 6 colon‐cancer cell lines. In colonic mucosa from 3 patients with ulcerative colitis it was not detected. The expression of MMP‐2 (72‐kDa type‐IV collagenase) mRNA was also investigated in the same tissue samples, and was detected in all samples, including cancerous and non‐cancerous tissue. Our data suggest that MMP‐7 is expressed in a tumor‐associated manner in colorectal cancers and may play a role in tumor progression.
Association of the antibodies against human papillomavirus 16 E4 and E7 proteins with cervical cancer positive for human papillomavirus DNA
Authors: Onda Takashi , Kanda Tadahito , Zanma Soichi , Yasugi Toshiharu , Watanabe Sumie , Kawana Takashi , Ueda Kuniaki , Yoshikawai Hiroyuki , Taketani Yuji , Yoshiike Kunito
Published: February 2007

Occurrence of the antibodies against human papillomavirus (HPV) 16 proteins E4 and E7 is specifically but independently associated with cervical cancer. To correlate HPV DNA and antibody data, we examined the biopsy specimens and sera, by polymerase chain reaction (PCR) and by ELISA, respectively, from 51 patients with cervical cancer (including 3 recurrent cases) and 22 with cervical intra‐epithelial neoplasia. Consensus primers for the LI region were used for PCR and bacterially expressed, purified fusion protein HPV‐16 E4 and non‐fusion protein HPV‐16 E7 were used for ELISA. HPV‐16 DNA and other HPV types were detected in 17 and 25, respectively, out of 51 cases of cervical cancer. Ten out of the 17 HPV‐ 16‐DNA‐ positives were positive either for anti‐E4 or for anti‐E7: positivities for anti‐E4, for anti‐E7, and for both were 6/17, 5/17 and 1/17 respectively. Three anti‐E7‐positives consisted of those for HPV‐33, ‐52 and ‐58 DNA, suggesting that limited cross‐reaction occurred between the HPV types. Among the HPV‐ 16‐DNA‐positive cases of cancer, lymph‐node or distant metastasis was recorded more frequently in the seropositives than in the seronegatives. Our results show that the HPV‐16 anti‐E4 or anti‐E7 occurs in some, but not in all, of the HPV‐16‐DNA‐positive cases, and support the hypothesis that the presence of the HPV‐16 antibodies can be used as a marker for possible metastasis.
Ectopic production of heparin‐binding growth factors and receptors for basic fibroblast growth factor by rat mammary epithelial cell lines derived from malignant metastatic tumours
Authors: Fernig David G. , Barraclough Roger , Ke Youqiang , Wilkinson Mark C. , Rudland Philip S. , Smith John A.
Published: February 2007

A rat mammary (Rama) epithelial cell line, Rama 704, derived from normal rat mammary gland does not possess any detectable cell‐surface receptors for basic fibroblast growth factor (bFGF), produces a barely detectable level of bFGF mRNA and does not contain detectable levels of bFGF‐like activity. Similar results have been obtained with the Rama 37 epithelial cells derived from benign tumours. However, 4 independently isolated epithelial cell lines derived from malignant rat mammary tumours and their metastases possess receptors for bFGF and contain between 2 ng and 9 ng heparin‐binding, growth‐stimulatory activity per 106 cells. The weakly metastatic Rama 600 cells possess high‐ and low‐affinity receptors for bFGF, (KD 20 pM and 8 nM, respectively), while the moderately metastatic Rama 800 cells possess only high‐affinity receptors (KD 40 pM). The moderately metastatic C18PLN and 267LU cells, derived from metastases arising from benign Rama 37 cells which had been transfected with DNA from the malignant Rama 800 cells, also possess only high‐affinity receptors (KD 36 pM and 80 pM, respectively). Our results show that within the Rama system there is a correlation between the appearance of heparin‐binding growth factors and of high‐affinity but not low‐affinity receptors for bFGF with the malignant phenotype.
The effects of tumour necrosis factor alpha on the vascular bed and blood flow in an experimental rat hepatoma
Authors: Naredi Peter L. J. , Lindnér Per G. , Holmberg Stig B. , Stenram Unne , Peterson Anders , Hafström Larsolof R.
Published: February 2007

The influence of TNF α on tumour growth rate has been attributed to its effects on the vascular bed and blood flow. The aim of our study was to investigate the effects of pharmacological doses of TNF α on the tumour vascular bed and to quantify blood flow in an experimental hepatoma during a more extended period after TNF‐α exposure than hitherto reported. In Lister rats, a syngeneic rat hepatoma was implanted on the dorsum of the right hind foot. TNF a was given i.v. The injection was repeated after 24 hr. Tumour blood flow was estimated before and 1, 24, and 96 hr after TNF‐α administration with the 133Xe‐washout technique. The passage of microspheres through the tumour vascular bed (non‐entrapment), as a measure of vascular occlusion, was estimated 4 and 96 hr after TNF‐α administration. Tumour growth rate was measured. The tumours were subjected to histological examination and the sensitivity to TNF α in vitro was tested. A reduction of tumour blood flow was observed in TNF‐α‐treated groups. Tumour growth rate was equally increased after 96 hr in both the TNF‐α groups as compared with controls. There was no significant change in non‐entrapment for the TNF‐a‐treated rats as compared with controls. Histology revealed extensive necrosis and thrombosis in tumours. TNF a had no effect on the viability of the cloned hepatoma cell line in vitro.
The effect of hypoxia on acquired drug resistance and response to epidermal growth factor in chinese hamster lung fibroblasts and human breast‐cancer cells in vitro
Authors: Kalra R. , Jones A‐M. , Kirk J. , Adams G. E. , Stratford I. J.
Published: February 2007

Prolonged hypoxia induced transient drug resistance in Chinese hamster lung fibroblasts. Previously hypoxic cells were resistant to adriamycin and resistant to etoposide. Complete recovery of etoposide sensitivity was observed following re‐aeration for 24 hr. A change in P‐glycoprotein expression was unlikely to contribute to the resistance caused by hypoxia, since adriamycin resistance was not reversed by verapamil. However, alteration in the plasma membrane structure may be involved, since previously hypoxic cells were resistant to extracellular superoxide radical generated by the addition of xanthine/ xanthine oxidase. In contrast, adriamycin sensitivity was not altered by hypoxia in 3 human breast‐cancer cell lines. MDA‐468 and MCF‐7/Adr differed in their response to EGF, independent of the presence of hypoxia. These results suggest that hypoxic‐stress‐induced drug resistance is not generalized.
Mutation and expression of the p53 gene in malignant melanoma cell lines
Authors: Weiss Jürgen , Schwechheimer Karl , Cavenee Webster K. , Herlyn Meenhard , Arden Karen C.
Published: February 2007

Three monoclonal antibodies (MAbs) against p53 protein (PAb 24o, DO‐I and Pab1801) were used to define the immunophenotype of 13 melanoma cell lines. Immunoreactions could be detected in 12 out of 13 cell lines by using the indirect immunofluorescence technique. In 7 of these the majority of cells displayed cytoplasmic staining whereas positive nuclei were detected in only a few cells. Two cell lines had predominantly nuclear reactivity, while the remaining 3 cell lines showed signals in both locations. Despite identical nuclear staining patterns, the 3 MAbs produced qualitatively distinct cytoplasmic immunoreactions. PAb240 and DO‐ I, which showed similar staining frequencies, appeared more sensitive in the detection of p53 protein than did PAb 1801. Immunoprecipita‐tions of lysates from each of the cell lines with MAbs DO‐I and 1801 (which bind to both wild‐type and mutant p53 species) detected 53–kDa proteins, whereas PAb240 (which recognizes the mutant conformation of the protein in this type of assay) detected 53–kDa proteins in only 4 cell lines. Nucleotide sequencing of exons 5 to 9 of TP53 in these latter cell lines showed that each has homozygous point mutations in the locus, whereas in the others no TP53 alterations were found. Three of the 4 mutations were C‐to‐T transversions, alterations possibly caused by damage from UV‐light. Our findings indicate that immunostaining with p53 antibodies, although common in malignant melanoma, results from the presence of mutant p53 protein in about 30% of the cases tested. Neither immunostaining with PAb240 nor the patterns of intracellular distributions of the signals are sufficient to detect TP53 mutations.
p53 Mutation in gastric cancer: A genetic model for carcinogenesis is common to gastric and colorectal cancer
Authors: Uchino Shinya , Noguchi Masayuki , Ochiai Atsushi , Saito Takao , Kobayashi Michio , Hirohashi Setsuo
Published: February 2007

Mutation of the p53 tumour‐suppressor gene in exons 5 through 8 was examined in 118 cases of gastric cancer (59 early gastric cancers and 59 advanced gastric cancers) using PCR‐SSCP (polymerase‐chain‐reaction‐single‐strand‐conformation polymorphism) analysis and direct sequencing. In early gastric cancer, mutations were found in 15 of 41 (37%) cases of the cohesive type, i.e., papillary adenocarcinoma, well to moderately differentiated tubular adenocarcinoma, and poorly differentiated adenocarcinoma with solid nests or focal tubular structures, but were not detected in 18 cases of the noncohesive type, i.e., signet‐ring‐cell carcinoma and poorly differentiated adenocarcinoma growing in a scattered manner. In advanced gastric cancer, 25 of 59 (42%) cases of the cohesive type had p53 mutation. No significant association was found between p53 mutation and other histopathological parameters such as macroscopic classification, lymph‐node involvement and depth of tumour invasion. Fifteen of 25 (60%) mutations in the advanced gastric‐cancer group were accompanied by allele loss at the p53 gene locus. Eighty‐three percent of mutations in early gastric cancer and 52% of mutations in advanced gastric cancer showed G:C‐to‐A:T transition, almost exclusively at CpG dinucleotide mutational hot spots, indicating that the spectrum of p53 mutation was similar to that of colorectal cancer. These data suggest that the p53 mutation occurs selectively in gastric cancer of the cohesive type from the intramucosal cancer stage.
Endocrine cells in colorectal adenocarcinomas: Incidence, hormone profile and prognostic relevance
Authors: De Bruïne A. P. , Wiggers Th. , Beek C. , Volovics A. , Von Meyenfeldt M. , Arends J. W. , Bosman F. T.
Published: February 2007

The occurrence of endocrine cells in 350 cases of colorectal adenocarcinoma was studied by immunohistochemistry for chromogranin A (CGA). The hormone profile of endocrine tumor cells, the correlation between endocrine differentiation and presence of other colorectal epithelial‐cell lineages and the prognostic relevance of endocrine differentiation in colorectal cancer were investigated. CGA‐positive tumor cells were found in 30% of cases, 21% showing moderate positivity and 9.0% extensive positivity. Of CGA‐positive tumors, 70% additionally produced neurohormones, mainly indigenous to normal colorectal epithelium: 55% showed immunoreactivity for glucagon‐like substances, 20% for serotonin and 10% for somatostatin, PYY and HCG. No immunoreactivity was found for various neurohormones not normally produced by colorectal endocrine cells. CGA‐positive tumors tended to be more aggressive than CGA‐negative tumors. Especially, tumors with extensive CGA positivity showed shorter survival, which was most apparent within Dukes' stage C. In multivariate analysis, extensive CGA positivity was an independent indicator of poor prognosis. CGA immunoreactivity significantly correlated with mucin production, but not with expression of secretory component (SC), a columnar‐cell marker. Mucin production significantly correlated with SC expression. Tumors positive for CGA but not for mucin and/or SC showed the worst prognosis. SC expression was a relatively favorable feature, and mucin‐producing tumors showed intermediate behavior.
Vase mini‐exon usage by ncam is not restricted to tumours of neuroectodermal origin
Authors: Patel K. , Culverwell A. , Rossell R. J. , Kemshead J. T. , Phimister E.
Published: February 2007

The neural cell adhesion molecule (NCAM) plays an important role in normal development. Many variants of NCAM are generated through post‐transcriptional and post‐translational modifications. These variants are tissue‐specific and their expression is developmentally regulated. NCAM is also re‐expressed in a number of human tumours, including neuroblastoma, rhabdomyosarcoma, Wilms' tumour and Ewing's sarcoma. We have characterized the NCAM variants associated with rhabdomyosarcoma. Polysialylated NCAMs are present in this tumour and, after neuraminidase treatment, they resolve into 2 bands of 140 and 120 kDa. These data were corroborated by Northern‐blot analysis where mRNA species of 6.7 and 5.5 kb are detected. These mRNA code for the 140‐ and 120‐kDa NCAM proteins respectively. PCR analysis shows that the previously described VASE mini‐exon is also present in NCAM found in rhabdomyosarcoma. The VASE mini‐exon, spliced at exon 7–8 junctions, has previously been detected in neural and heart NCAM, as well as in NCAMs found in human small‐cell lung carcinoma (SCLC). DNA sequencing confirmed that the VASE mini‐exon in rhabdomyosarcoma is identical to that found in neuroblastoma and SCLC.