bluuebunny/crossref_metadata_2025 huggingface dataset:
Membrane association and shedding of the GPI‐anchored Ca‐MOv18 antigen in human ovary carcinoma cells
Authors: Miotti S. , Alberti S. , Facheris P. , Mantovani L. , Fornaro Mara , Stella Manuela , Ménard S. , Canevari S. , Colnaghi M. I.
Published: February 2007

The antigen recognized by the MOvl8 MAb (Ca‐MOvl8) was recently shown to be a glycosylphosphatidylinositol (GPI)‐linked protein. In this report we show that GPI‐anchorage is not limited to IGROVI cells nor to other ovary carcinoma cell lines, but Ca‐MOvl8 was also found to be sensitive to phosphatidylinositol‐specific phospholipase C (PI‐PLC) treatment on fresh ovarian cancer cells. Furthermore, we found a heterogeneous sensitivity of Ca‐MOv18 to PI‐PLC cleavage, not only among the different cells studied but also in different experiments performed on the same cell line, during extended periods of time in culture. Sensitivity to PI‐PLC cleavage was determined by immunofluorescence on live cells and by double‐determinant radioimmunoassay of the antigen released in the supernatant. The specificity of the PI‐PLC cleavage was demonstrated as follows: (a) TX114 solubilized Ca‐MOv18 shifts from the detergent to the aqueous phase after treatment with PI‐PLC; (b) on membrane preparations, PI‐PLC specifically released a fraction of the antigen, which is distinct from the weakly associated form released by high‐salt treatment; (c) Ca‐MOvI8 from IGROVI expressed the cross‐reacting determinant (CRD), which is characteristic of GPI‐linked molecules. The absence of CRD expression on the spontaneously released protein and the possibility of artificially inducing antigen shedding during the resynthesis of Ca‐MOvI8 which follows bacterial PI‐PLC treatment are interesting points which need to be further investigated in order to understand the physiology of the Ca‐MOvI8 tumor antigen.
Human papillomavirus (HPV) infection, hiv infection and cervical cancer in Tanzania, East Africa
Authors: Meulen J. Ter , Eberhardt H. C. , Luande J. , Mgaya H. N. , Chang‐Claude J. , Mtiro H. , Mhina M. , Kashaija P. , Ockert S. , Yu X. , Meinhardt G. , Gissmannl L. , Pawlita M.
Published: February 2007

The presence of HPV‐DNA was determined in tumor biopsies of cervical‐cancer patients and in cervical swabs of non‐cancer patients from Tanzania, East Africa, by Southern blot hybridization and/or PCR. HPV types 16 and 18 were detected in 38% and 32%, respectively, of 50 cervical‐carcinoma biopsies. A consensus primer PCR capable of detecting a broad spectrum of HPV types revealed the presence of HPV‐DNA in 59% of 359 cervical swabs of non‐cancer patients. Type‐specific PCR showed that types 16 and 18 accounted for 13.2% and 17.5%, respectively, of all HPV infections. Therefore we concluded that HPV 18 is more prevalent in Tanzania than in any other geographical location so far reported. The strongest risk factors for the presence of any HPV‐DNA in the 359 female non‐cancer patients were young age and HIV infection. The epidemiology of HPV types 16 and 18 was found to differ from that of other HPV types, being associated in univariate analysis with trichomonas vaginalis infection, marital status (single/divorced), age at first intercourse, and young age at menarche. However, young age at menarche accounted for most of the effects of all other variables in multivariate analysis. Of the non‐cancer patients, 12.8% had antibodies against HIV I (no patient being severely symptomatic), and HIV infection was highly correlated with the presence of HPV‐DNA, especially types 16 and 18. While HPV‐DNA of any type was detectable 1.4‐fold more often in HIV‐positive patients than in HIV‐negative patients, evidence of an infection with HPV types 16 or 18 was found 2.2‐fold more often in the HIV‐positive patients. The HIV‐positive women did not show an increased rate of cervical cytological abnormalities as assessed by PAP staining of a single cervical smear, the overall rate of abnormalities being 2.8%. Furthermore, the age‐adjusted prevalence of HIV antibodies was found to be considerably lower in 270 cervical‐carcinoma patients (3% HIV‐positive) in comparison with non‐cancer patients. Thus there was no association observable between the prevalence of HIV infections and the frequency of cervical cytological abnormalities or cervical cancer in the setting of this cross‐sectional study.
Expression of Cx26, Cx32 AND Cx43 gap junction proteins in normal and neoplastic human tissues
Authors: Wilgenbus Klaus K. , Kirkpatrick Charles J. , Knuechel Ruth , Willecke Klaus , Traub Otto
Published: February 2007

This report concerns the expression of the gapjunction proteins Connexin (Cx)26, 32 and 43 in different malignant and nonmalignant human tissues. Affinitypurified polyclonal antibodies against Cx26, 32 and 43 were used for immunohistochemical as well as immunoblot analysis. Cx32, the major gapjunction protein in rat and mouse liver, was detected in human liver and kidney. By contrast, Cx43 was expressed in epithelial and mesenchymal tissues and Cx26 was detected in different epithelia. Whereas all of the benign tumors studied, and some malignant ones, showed stable expression of gapjunction proteins, breast cancer, renalcell cancer and sarcomas showed a significant decrease in gapjunction proteins as opposed to normal tissue. Cx43, not detected in human normal liver, was found in human hepatocellular carcinoma and Cx26, not detected in human adult skin, was observed in tissue samples of basalcell carcinoma. In immunoblot analysis, Cx32 antibodies recognized a 27‐kDa protein in human liver and hepatocellular carcinoma. A 43‐kDa polypeptide was detected in human kidney, renalcell carcinoma, normal breast, connective tissue of invasiveduct carcinoma of the breast and hepatocellular carcinoma. © 1992 Wiley‐Liss, Inc.
Basement membrane collagen‐IV synthesis in colorectal tumours
Authors: Hewitt Robert E. , Powe Desmond G. , Carter G. Ian , Turner David R. , Price Janet E.
Published: February 2007

Many studies suggest that increased proteolysis accounts for the epithelial basement membrane (EBM) breaks commonly seen in carcinomas. As failure to produce or maintain EBM may also be important, we chose to investigate synthesis of basement membrane collagen‐IV in human colorectal carcinomas. First, to determine the cellular origin of EBM collagen‐IV, speciesspecific antibodies were used to analyse caecal xenografts of 4 different human colorectalcarcinomaderived cell lines. The results of this study suggest an exclusively stromal cell origin for EBM collagen‐IV. Next, the distribution of periglandular myofibroblasts in carcinomas was examined, since in normal mucosa their location and ultrastructural features suggest that they play a role in EBM maintenance. They were generally abundant in normal mucosa and adenomas, but sparsely distributed in carcinomas, particularly at the invasive periphery where EBM collagen‐IV immunostaining is most deficient. Finally, the in situ hybridization technique was used to define cell populations synthesizing collagen‐IV. In normal mucosa, no collagen‐IV mRNA was detected in any component, while in carcinomas, the mRNA was clearly detectable in vascular endothelial cells but not in any other cell type. Increased vascular collagen‐IV production in carcinomas may be at least partly due to tumourinduced angiogenesis, since new bloodvessel formation requires the synthesis of new vascular basement membranes. © 1992 Wiley‐Liss, Inc.
Alcohol consumption as A risk factor for hepatocellular carcinoma in urban Southern African blacks
Authors: Mohamed Abdulla E. , Kew Michael C. , Groeneveld Hendrik T.
Published: February 2007

Our purpose was to ascertain whether alcohol abuse is a risk factor for the development of hepatocellular carcinoma in urban southern African blacks and, if so, to relate alcohol consumption to other possible risk factors such as persistent hepatitis‐B‐virus infection, smoking, male sex, in this subpopulation. A prospective, hospitalbased, casecontrol format involving 101 patients with hepatocellular carcinoma and 101 controls was used. The mean age of the patients was 53.7 ± 1.85 years and the male:female ratio 3.2:1. An increased risk was found, but only in urban men over the age of 40 years who habitually drank more than 80 g of ethanol daily. The risk remained after adjusting for chronic hepatitis‐B infection, smoking, and sex (odds ratio 4.4, 95% confidence interval 1.3 to 16.6; p = 0.003). Smoking proved not to be a risk factor, either alone or in concert with alcohol consumption. Hepatitis‐B infection was confirmed as a major risk in younger men and in women, but in urban men over the age of 40 years alcohol abuse was a greater risk. Current hepatitis‐B infection and alcohol abuse were additive risks. © 1992 Wiley‐Liss, Inc.
An attempt to define sets of cooperating genetic alterations in human breast cancer
Authors: Cheickh Meryem Ben , Rouanet Philippe , Louason Geneviève , Jeanteur Philippe , Theillet Charles
Published: February 2007

The extent and the variation of losses of genetic material were examined in a series of 191 human breast cancers by means of a set of 18 polymorphic DNA probes, specific of 7 chromosomal arms (1p, 1q, 3p, 11p, 13q, 17p and 18q) known to be frequently affected by allele losses. Frequencies of losses of heterozygosity ranged from a low of 3.5% (chromosome 13q) to a high of 27% (chromosome 3p). The number of sites involved in breast cancer added to the frequent occurrence of concomitant losses at several chromosomal arms within the same tumor suggest cooperative effects of these LOHs. We were therefore interested in assessing the existence of preferential associations between sets of LOHs in our panel of tumors. Statistically significant associations were found between LOHs at chromosomes Ip and 17p, and between LOHs at chromosomes 11p and 17p. Furthermore, since all the tumors presently studied had previously been analyzed for protooncogene amplification at 5 distinct chromosomal sites, we tested for associations between LOH and DNA amplification. Such associations were indeed observed as exemplified by the correlations observed between the LOH at 11p and amplification of the erbB2 gene and LOH at 17p and the amplification of the fig gene. The only correlation with clinicopathological parameters that could be observed linked the occurrence of LOHs on 11p with recurrent breast cancer (p = 0.015). Sets of several LOHs or LOHs and gene amplifications could not be significantly related to any marker of tumor aggressiveness. © 1992 Wiley‐Liss, Inc.
DNA flow cytometry, nuclear morphometry, mitotic indices and steroid receptors as independent prognostic factors in female breast cancer
Authors: Eskelinen Matti , Lipponen Pertti , Papinaho Sari , Aaltomaa Sirpa , Kosma Veli‐Matti , Klemi Pekka , Syrjänen Kari
Published: February 2007

Clinical features, 8 histological variables, 7 nuclear morphometric variables, 2 mitotic indices, oestrogen‐receptor (ER) and progesterone‐receptor content (PR), DNA ploidy and S‐phase fraction (SPF) were entered in a Cox's model to assess their independent predictive value in 216 breast‐cancer patients followed up for over 9 years. In the whole series, histological type (p = 0.007), volume‐corrected mitotic index (M/V index) (p = 0.01), axillary‐lymph‐node (pN) status (p = 0.024) and the year of treatment (p = 0.045) predicted independently the recurrence‐free survival (RFS). In a sub‐analysis including SPF (n = 148), the year of treatment (p = 0.003), tumour diameter (p = 0.004), SPF (p = 0.022) and nuclear pleomorphism (p = 0.056) independently predicted the RFS. In a Cox's analysis of the whole series, tumour diameter (p < 0.001), pN status (p = 0.001), PR status (p = 0.002) and the year of treatment (p = 0.021) were independent predictors of survival. In a separate analysis including also SPF (n = 148), tumour diameter (p < 0.001), SPF (p = 0.003), pN status (p = 0.008) and the year of treatment (p = 0.015) proved to be independent prognostic factors. The results show that tumour diameter, pN status, M/V‐index, histological type, SPF and PR status comprise a sufficient combination of prognostic factors in female breast cancer. In pN patients, age and SDPE may be of additional prognostic significance. The prognostic scores combining the independent prognostic variables reflecting both the proliferative rate and metastatic potential of the tumours are accurate predictors of the RFS and overall survival.
Human papillomavirus (hpv) DNA in penile carcinomas and in two cell lines from high‐incidence areas for genital cancers in Africa
Authors: Tornesello Maria L. , Buonaguro Franco M. , Beth‐giraldo Elke , Kyalwazi S. K. , Giraldo Gaetano
Published: February 2007

Biopsies of 13 penile cancers (PC), from patients living in regions of Uganda with a high incidence of genital cancers, were studied for the presence, molecular characteristics and physical state of DNA related to that of human papillomavirus (HPV) types 6, 11, 16, 18, 31 and 33. HPV DNA sequences were detected in all PC specimens by dot/Southern blot analyses and by gene amplification of DNA sequences highly conserved among several HPVs. HPV 16 DNA sequences were found in one PC; DNA sequences with low homology to HPV 16 or HPV 18 were present in all other samples. Viral DNA is primarily integrated in the cellular DNA. To isolate and characterize a possible highly oncogenic HPV, a genomic library of the DNA extracted from the PC‐8 biopsy has been constructed in the EcoRI arms of the EMBL4 phage. A single phage containing 8.30‐kb HPV16‐related sequences has been identified and the 3 segments of 0.45, 0.65 and 7.2 kb, released by EcoRI digestion, have been independently subcloned in pUCI8 for further analysis.
Studies of clonal cell lines developed from primary breast cancers indicate that the ability to undergo morphogenesis IN vitro is lost early in malignancy
Authors: Shearer Moira , Bartkova Jirina , Bartek Jiri , Berdichevsky Feodor , Barnes Diana , Millis Rosemary , Taylor‐Papadimitriou Joyce
Published: February 2007

In an attempt to obtain cell lines from the different components found in primary breast cancers, we used a low‐calcium medium to culture epithelial cells of mixed phenotype and a recombinant T antigen containing retrovirus to immortalize cells found in these cultures. In each case, the histology of the sample used for culture was examined in detail and the best growth was obtained from samples associated with a substantial in situ or benign component and from lobular rather than ductal carcinomas. Clonal cell lines were developed from each of 4 tumours: I infiltrating ductal (tumour number 2), 2 infiltrating lobular (tumours 3 and 5) and I mucoid (tumour 6). To try to identify the phenotype and origin of the cell lines, immunohis‐tochemical markers, histological analysis of tissue sections and behavioural markers were used. All the cell lines expressed mainly luminal epithelial cell markers, but the basal epithelial keratin, keratin 14, was also expressed homogeneously or heterogeneously. Growth in agar was seen with some but not all cell lines derived from only I tumour (tumour 5) and tumour development in nude mice was observed (with low efficiency) with cell lines from only I tumour (tumour 6). The data suggest that the cell lines obtained from the infiltrating ductal carcinoma (tumour 2) developed from cells cultured from the associated benign component, while the cell lines from tumours 3, 5 and 6 may each have developed from a cell in an early stage of malignancy. When tested for their ability to undergo morphogenesis on extracellular matrix components, celi lines from tumour 2 made well‐developed ductal‐alveolar‐like structures, while those from the other tumours did not.
Unique expression of integrin fibronectin receptors in human neuroblastoma cell lines
Authors: Yoshihara Takao , Esumi Noriko , Humphries Martin J. , Imashuku Shinsaku
Published: February 2007

Cultured human neuroblastoma cells can be classified morphologically into 3 types: neuroblastic (N), intermediate (I) and substrate adherent (S). Neuroblastoma cells of all types were found to attach and display distinct morphological characteristics on fibronectin, with S‐type cells attaching better than N‐type cells. Studies of the expression of integrin fibronectin receptors (α3βI, α4β I, α5β I and αVβ I) were carried out using a total of 26 morphologically distinct cell lines and their subpopu‐lations. Fluorescence‐activated cell sorting (FACS) analysis and immunoprecipitation revealed that all S‐type cells expressed abundant α5β I, while N‐type cells barely expressed this molecule. Although α3β I expression of S‐type cells was also higher than that of N‐type cells, some N‐type cells had significantly increased levels of this molecule. α4β I was found to be randomly expressed. All cell lines tested expressed αVβ I. Human neuroblastoma cells, the majority of which are N‐type cells with very low α5β I expression, are also contrasted with other childhood cancer cells (rhabdomyosarcoma, Ewing's sarcoma, and glioma), all of which expressed high levels of α5β I. The characteristic expression of integrin fibronectin receptors may account for the clinically unique tumor behavior, and the immunohistochemical staining for integrins may become a useful alternative to conventional histology in differential diagnosis and a marker for prognosis in neuroblastoma.
Dipyridamole enhances an anti‐proliferative effect of interferon in various types of human tumor cells
Authors: Suzuki Nobuo , Oiwa Yoko , Sugano Isamu , Inaba Noriyuki , Sekiya Souei , Fukazawa Ichio , Yoshida Jun , Takakubo Yoshiaki , Isogai Eriko , Saito‐Ebihara Mitsue
Published: February 2007

The anti‐proliferative activity of human interferon (HulFN) was enhanced by dipyridamole, 2,6‐bis‐(diethanolamino)‐4,8‐dipiperidinopyrimido‐[5,4‐d]‐pyrimidine, when tested against various human tumor cell lines, including KT (breast carcinoma), PLC/PRF/5 (hepatoma), MGC‐1, U251‐SP and T98 (glioma), HAC‐2 and SHIN‐3 (ovarian carcinoma), and MM‐1CB (melanoma). The enhancement occurred irrespective of the kind of HulFN used (α, β or γ) and the original degree of susceptibility of the cells to HulFN. Even low doses down to 0.01 μM of dipyridamole that had no intrinsic anti‐proliferative activity could enhance the effect of HulFN. The enhancement of HulFN effects seems not to be caused by induction of HulFN production, because neither anti‐viral activity nor HulFN antigens were detected in culture medium in cells treated with dipyridamole. Mopidamole, a derivative of dipyridamole lacking one piperidine residue, produced little enhancement of the effects of HulFN. Among ovarian cancer cell lines tested, the enhancement of the activity of HulFN by dipyridamole for HAC‐2 and SHIN‐3 cells was equivalent to or greater than that for 3 chemotherapy agents (adriamycin, vincristine, and a camptothecin derivative). However, neither HOC‐21 ovarian cancer cells nor HEC‐1 endometrial adenocarcinoma cells were susceptible to any combinations. When MGC‐1, U251‐SP, and HAC‐2 cells were injected into nude mice, the growth of tumors was more markedly inhibited by the subcutaneous administration of HulFN in combination with oral administration of dipyridamole than by the HulFN alone. Thus, this combination therapy seems to be worth trying for human cancer, although the enhancement of the effects of HulFN by dipyridamole varied among the cell lines examined.
Regulation of surface‐differentiation molecules by epidermal growth factor, transforming growth factor alpha, and hydrocortisone in human mammary epithelial cells transformed by an activated c‐Ha‐ras proto‐oncogene
Authors: Basolo F. , Serra C. , Ciardiello F. , Fiore L. , Russo J. , Campani D. , Dolei A. , Squartini F. , Toniolo A.
Published: February 2007

Spontaneously immortalized human mammary epithelial cells MCF‐IOA were transfected with an activated c‐Ha‐ras oncogene. Transfected cells (MCF‐IOT) acquire a malignant pheno‐type, as already reported. Studies of l‐2′‐deoxyuridine incorporation in cultures given graded doses of hydrocortisone (HC), cholera toxin (CT), epidermal growth factor (EGF), and transforming growth factor alpha (TGF‐α) showed that though MCF‐IOT had become almost independent on exogenous EGF and TGF‐α, they continued to respond to the synergistic effect of HC and CT plus EGF. Both lines were phenotypically characterized with an immunoradiometric assay in live cells. Expression of MHC class‐I molecules, human milk‐fat‐globule‐1 antigen, and EGF receptor was reduced in ras‐transfected cells, although other differentiation markers were unchanged. Exogenous EGF down‐regulated the expression of functional EGF‐R, selectively in transformed cells. TGF‐α failed to modulate EGF‐R. In contrast, HC strongly stimulated the expression of EGF‐R while depressing MHC class‐l molecules. Thus, it appears that in vivo HC may co‐operate with TGF‐α and EGF in promoting the growth of transformed mammary cells. This hormone might also favor the escape from immune surveillance by reducing the expression of surface differentiation markers.
A single human colonic adenoma cell line can be converted IN VITRO to both A colorectal adenocarcinoma and A mucinous carcinoma
Authors: Paraskeva Christos , Hague Angela , Rooney Nicholas , Williams Ann C. , Harper Sara J. , Hanlon Katherine A. , Atkinson Rebecca J. , Corfield Anthony P.
Published: February 2007

In a previous study, using a chemical carcinogen, we converted in vitro a non‐tumorigenic cell line derived from a human colorectal diploid adenoma, designated PC/AA, into a tumori‐genic cell line which, when inoculated into athymic nude mice, produced progressively growing adenocarcinomas. We now report that continuous in vitro passage of the PC/AA adenoma cell line resulted in its spontaneous transformation to a mucinous carcinoma with a modal karyotype of 51, XY, + i( Iq), +8, +9, +13, +i(13q), ‐21, +mar. These studies show that a single adenoma can be converted along 2 independent pathways, giving rise to either a mucinous carcinoma or an adenocarcinoma, and provide further experimental evidence for the adenoma‐carcinoma sequence. Cytogenetic changes which occur along both pathways to tumorigenicity include abnormalities of chromosome I and multiple copies of chromosome 13. These abnormalities may be important in tumour development and progression in colorectal carcinogenesis.
Quantitative study of the growth of experimental hepatic tumors in rats by using magnetic resonance imaging
Authors: Qin Yunfeng , Van Cauteren Marc , Osteaux Michel , Willems Gérard
Published: February 2007

Precise estimation of the volume and growth rate of hepatic metastases would represent an important step forward not only in clinical oncology but also for the evaluation of experimental treatments in animal models. In the present study, an original method of volumetry of hepatic metastatic tumors in vivo has been tested in rats using magnetic resonance imaging (MRI). Three different hepatic tumor models mimicking liver metastases were established in syngeneic BDIX rats by injection of DHD/KI2 rat colon cancer cells either directly under the liver capsule or via the portal system. The liver tumor volumes were estimated in vivo by using MR imaging of the liverand summing the individual tumor volumes in the sequential MR liver sections. The values of the tumor volumes measured by MRI were compared with those determined by a classical method of water displacement in vitro after killing the animals and excising the tumors. At 3 weeks after tumor implantation, liver tumors as small as I mm in diameter could be detected by MRI. The difference between the tumor volumes estimated by MRI in vivo and those measured by water displacement in vitro was 9% for single liver tumors and 16% for multiple liver tumors. Close correlation between the values of the tumor volumes measured by MRI and those determined by water displacement was observed in solitary liver tumors (r = 0.985, p < 0.01) as well as in multiple liver tumors (r = 0.985, p < 0.01), indicating the high accuracy of MRI volumetry for liver tumors. Estimation of the liver tumor volumes by MRI in the same animals at successive time intervals made it possible to construct tumor growth curves and to calculate tumor growth parameters. These data suggest that MRI volumetry represents an effective means of evaluating the efficacy of experimental treatments in small animals and may have potentially important applications in clinical patients.
c‐myc amplification is an independent prognostic factor in postmenopausal breast cancer
Authors: Borg Åke , Baldetorp Bo , Fernö Mårten , Olsson Håkan , Sigurdsson Helgi
Published: February 2007

The c‐myc proto‐oncogene was analyzed in 311 cases of primary breast cancer, in 8% of which it was found to be amplified, usually at moderately increased copy number (2‐5 copies). The adjacent pvt gene was co‐amplified with c‐myc in all tumors analyzed. C‐myc amplification was significantly correlated to a high S‐phase fraction and to amplification of the c‐erbB‐2 proto‐oncogene. Weak relationships were found between c‐myc amplification and the presence of lymph‐node metastasis, advanced stage, DNA non‐diploidy and premenopausal status, but not tumor size, estrogen receptor or progesterone receptor status, or int‐2 amplification. C‐myc amplification, and especially a high gene copy number (>5 copies), was significantly related to early recurrence and death in breast cancer, a relationship seen in both the lymph‐node‐negative and node‐positive subcategories. A particularly strong correlation with poor clinical outcome was seen in postmenopausal patients (p > 0.0005), an association which persisted in multivariate survival analysis. We conclude that the activation of c‐myc is indeed associated with rapidly growing and progressive breast cancer. Gene amplification, on the other hand, is relatively infrequent and occurs mostly at low copy number, implying that tumors are heterogeneous with respect to cell clones harboring c‐myc amplification. An immunohistochemical assessment would more accurately illustrate the importance of c‐myc activation in human breast cancer. However, the obvious instability of the c‐myc transcript and translate suggests that c‐myc is not a suitable prognostic marker for routine purposes. © 1992 Wifey‐Liss, Inc.
Study of fibronectin mRNA in human laryngeal and ectocervical carcinomas by IN SITU hybridization and image analysis
Authors: Moro Laura , Colombi Marina , Tosatti Maria Pia Molinari , Barlati Sergio
Published: February 2007

The expression of fibronectin (FN) mRNA was studied in histological sections of surgical biopsies from human laryngeal and ectocervical invasive carcinomas of different grading stages by in situ hybridization and image analysis. This approach made it possible to identify the cell types synthesizing FN mRNA in the tissue sections and to compare semi‐quantitatively the FN mRNA levels expressed in the different specimens. The carcinoma cells synthesized low levels of FN mRNA, comparable to those detected in control epithelia and connective‐tissue fibroblasts. Well‐differentiated (G1) laryngeal and ectocervical carcinomas induced the synthesis of FN mRNA—to levels 7 to 13 times higher than in control connective tissues—in the stromal fibroblasts surrounding the tumors. In carcinoma samples analysed, the amount of FN mRNA detected in the stroma decreased in relation to tumor grading (from G1 to G3) and the stromal destruction. FN mRNA was not detectable in the endothelial cells of venules while it was present in large amounts in those surrounding the capillaries present in the stroma. These data indicate that FN, usually observed around carcinomas, is produced by stromal fibroblasts, which are induced to express FN mRNA, presumably in response to diffusible factors produced by the tumor cells, and/or by endothelial cells of the infiltrating capillary vessels. The induction of FN mRNA, inversely proportional to the tumor grading, may be useful in evaluating the invasion potential of the tumor. © 1992 Wifey‐Liss, Inc.
Loss of heterozygosity for distal markers on 22q in human gliomas
Authors: Rey Juan A. , Bello M. Josefa , Jimenez‐lara Ana M. , Vaquero Jesus , Kusak M. Elena , de Campos José M. , Sarasa José L. , Pestana Angel
Published: February 2007

Loss of constitutional heterozygosity as determined through the analysis of restriction‐fragment‐length polymorphism (RFLP) on tumoral and constitutional DNA has proven to be helpful to delimit the location of tumor‐suppressor genes in the human genome. In malignant gliomas this approach indicates that chromosomes 9p, 10, 17p, and 22 may contain genes of this category involved in its origin and/or progression. Regarding chromosome 22, the data so far provided by molecular studies confirmed those previously reported by cytogenetic studies, suggesting the existence of a sub‐group of malignant gliomas characterized by monosomy of this chromosome. However, the precise location of the putative glioma suppressor gene on chromosome 22 remains ambiguous. We have performed a combined cytogenetic and RFLP study on a series of 31 gliomas, looking for structural abnormalities of this chromosome. In 3 instances, terminal deletions of the long arm of chromosome 22 were observed by both methodologies, suggesting that the band q13 region distal to the D22S80 marker might be the critical domain non‐randomly involved in tumor suppression of gliomas. © 1992 Wifey‐Liss, Inc.
Cytostatic activity of new synthetic anti‐tumor aza‐alkyllysophospholipids
Authors: Sidoti Carole , Principe Paola , Vandamme Benedicte , Broquet Colette , Braquet Pierre
Published: February 2007

Alkyllysophospholipids are analogues of the naturally occurring 2‐lysophosphatidylcholine which have been reported to have selective in vitro/in vivo anti‐tumor activity. Their anti‐proliferative effect has been found against a variety of animal and human tumor cell lines. We have characterized the cytostatic activity of 2 newly synthetized aza‐alkyllysophospholipids (AALPs), the BN52205 and the BN52211, on a human tumor cell line derived from a colon adenocarcinoma, the HT29. We used 3 different flow cytometric approaches to study which phase of the cell cycle was the most sensitive to the anti‐proliferative activity of the 2 AALPs. By applying the biparametric analysis of 5′‐bromo‐2‐deoxyuridine incorporation vs. DNA content we have been able to demonstrate that the 2 AALPs do not interfere with the S phase of the cell cycle. The simultaneous measurement of total nuclear protein vs. DNA content in isolated HT29 nuclei enabled us to exclude a block in the M phase of the ee cycle. Finally, stathmokinetic analysis enabled us to show that cytostatic activity of the 2 new AALPs is characterized by multiple “terminal points” as the drugs' action results in a G1 block, in a slow‐down of the transition from late S to G2 followed by an accumulation of HT29 cells in the G2 phase of the cell cycle. © 1992 Wifey‐Liss, Inc.
Modulation of neuroblastoma cell differentiation by the extracellular matrix
Authors: Matsushima Hiroshi , Bogenmann Emil
Published: February 2007

The effect of a complex in vitro synthesized extracellular matrix (ECM) and its components on growth and phenotypical differentiation of a human neuroblastoma (NB) cell line (HTLA230) was investigated. Rat smooth‐muscle‐cell (R22CIF)‐derived ECM composed of collagen, glycoproteins, and glycosaminoglycans (GAGs) promoted spontaneous neurite outgrowth of HTLA230 cells but did not alter their growth kinetic or cloning efficiency as compared with cells seeded onto gelatin‐coated dishes. The matrix significantly enhanced, quantitatively and qualitatively, the responsiveness of HTLA230 cells to retinoic acid (RA), and a substantially reduced growth rate was observed in the presence of RA with cells grown on the ECM. Biochemical modification of the composition of the R22CIF‐matrix by trypsin digestion and/or high‐salt extraction (4 M guanidinium) demonstrated that the ratio of chondroitin sulfate to hyaluronic acid (HA) present in the ECM determines the capacity of the matrix to promote NB differentiation. A human fibroblast (T‐1)‐derived ECM, which has a biochemical composition of the GAG component similar to that of the trypsinized R22 CIF‐matrix, but which has a high amount of glycoproteins, confirmed these results. Nerve‐growth‐factor (NGF)‐induced differentiation in a variant HTLA 230 cell line was inhibited when cells were grown on an ECM with a low ratio of chondroitin sulfate/HA. The composition of the ECM thus modulates the responsiveness to various differentiation‐inducing agents and alters the phenotype of NB cells. © 1992 Wifey‐Liss, Inc.
Phosphorylation and estradiol binding of estrogen receptor in hormone‐dependent and hormone‐independent GR mouse mammary tumors
Authors: Migliaccio Antimo , Pagano Michele , de Goeij Cornelis C. J. , Domenico Marina Di , Castoria Gabriella , Sluyser Mels , Auricchio Ferdinando
Published: February 2007

The effect of phosphorylation on the hormone‐binding capacity of the estrogen receptor (ER) was investigated in hormone‐dependent (HD) and hormone‐independent (HI) mammary carcinomas of GR mice. Tumor cytosols were incubated with ATP under conditions previously used to study the tyrosine kinase which confers hormone binding to phosphatase‐treated or in vitro‐synthesized ER. The ATP‐dependent increases in hormone‐binding capacity of 8 out of 20 HI tumors ranged from values of 23 to 124 fmol/mg cytosol protein. The enhancement by ATP of hormone binding to ER was significantly less marked in HD and HR tumors than in HI tumors. In only 3 out of 13 HD and HR tumors was an increase ranging from 15 to 20 fmol/mg protein detected. Analysis by Scatchard plot of estradiol binding to ER showed that cytosol incubation of HI tumors with ATP markedly increased the hormone binding without any change in affinity. The data suggest that ER of HI tumors is less phosphorylated in vivo than the ER of HD/HR tumors, so that the receptor of HI tumors is more susceptible to γ32P‐ATP phosphorylation and ATP‐induced hormone binding in vitro. Western blot of ER with antiphosphotyrosine antibody showed that, in HI tumors, the large ATP‐induced increase in hormone binding to ER was associated with phosphorylation on tyrosine of the receptor itself. Our findings indicate that the process of activation‐inactivation of binding through tyrosine‐phosphorylation/ phosphotyrosine‐dephosphorylation of ER observed in estrogen target tissues is altered in some HI mammary tumors. © 1992 Wifey‐Liss, Inc.
Malignant transformation of NIH‐3T3 cells after subcutaneous co‐injection with a reconstituted basement membrane (matrigel)
Authors: Fridman Rafael , Sweeney Thomas M. , Zain Mona , Martin George R. , Kleinman Hynda K.
Published: February 2007

NIH‐3T3 cells are non‐tumorigenic when injected into athymic mice. If these cells are mixed with an extract of basement‐membrane proteins (matrigel) and injected s.c., they form locally invasive and highly vascularized tumors. Cells cultured from the NIH‐3T3‐matrigel‐induced tumors showed a transformed phenotype and lacked contact inhibition. When cultured in a gel of matrigel, they proliferated and formed branched and invasive colonies. In contrast, the parental NIH‐3T3 cells cultured on matrigel remained as cell aggregates and were not invasive. I.V. injections of the tumor‐derived NIH‐3T3 cells produced many colonies on the surface of the lungs, whereas the parental NIH‐3T3 cells were not metastatic. Zymographic analysis of the conditioned media obtained from both the tumor‐derived and parental NIH‐3T3 cells demonstrated higher amounts of the 72‐kDa gelatinase (type‐IV collagenase) enzyme in the tumor‐derived cells. Also, tumorderived NIH‐3T3 cells, but not parental NIH‐3T3 cells, secreted the 92‐kDa type‐IV collagenase. These studies suggest that the interaction of pre‐malignant NIH‐3T3 cells with extracellular matrix components may contribute to the process of tumor progression. © 1992 Wifey‐Liss, Inc.
Putative invasion‐specific proteins in mouse T‐cell hybridomas that differ in invasive and metastatic potential
Authors: La Rivière Geertje , Schipper Cor A. , Klein Gebbinck Jacqueline W. T. M. , Koch Guus , Kuhn Lotte , Lefkovits Ivan , Roos Ed
Published: February 2007

Fusion of invasive, activated T‐lymphocytes with non‐invasive BW5147 T‐lymphoma cells mainly yields highly invasive (HI), highly metastatic T‐cell hybridomas. In addition, several noninvasive (NI), non‐metastatic hybrids have been obtained, probably due to loss of involved gene(s) by chromosome segregation. Here we have compared a panel of HI and NI hybrids in a search for proteins specifically expressed by either cell type. MAbs were raised against HI hybrids, but out of more than 1,000 none bound exclusively to HI cells. Furthermore, polyclonal rat, rabbit and chicken antisera did not immunopre‐cipitate specific proteins from total lysates, and the expression of 18 (T‐cell) surface markers did not correlate with invasiveness. These results indicated that the number of differences between HI and NI hybridomas was surprisingly small. This notion was confirmed by 2‐dimensional gel electrophoresis. Among 1,000 detectable spots, we found only 2 clear‐cut differences between HI and NI T‐cell hybridomas, whereas multiple differences were found between individual hybrids. One protein (p130) was expressed at much higher levels by HI than by NI hybrids in this panel, whereas the other (p 15) was only seen in NI hybrids. These proteins are primary candidates for a role in invasion. © 1992 Wifey‐Liss, Inc.
Transformation of rodent fibroblasts by herpes simplex virus: Presence of morphological transforming region 1 (MTR 1) is not required for the maintenance of the transformed state
Authors: Bauer Georg , Kahl Susanne , Sawhney Iva Singh , Höfler Petra , Gerspach Ralph , Matz Bertfried
Published: February 2007

Studies on the mechanisms of transformation of mammalian cells by herpes simplex virus (HSV) in vitro have been prevented so far by the extremely low transformation frequencies obtained in monolayer culture. Here we present a transformation system that relies on the direct seeding in soft agar of infected single cells, thus avoiding negative interactions between normal and transformed cells. We took advantage of HSV‐1 temperature‐sensitive mutants at the UL9 locus, which codes for a DNA‐binding protein necessary for viral DNA replication. At the non‐permissive temperature, viral DNA synthesis and late gene expression are prevented. Viral gene expression is restricted to immediate early and early genes. Induction of transformation was highly efficient in our one‐step transformation system. It depended on intact viral particles and viral DNA. Immediate early and/or early viral gene expression was sufficient to induce transformation. Colonies were stably transformed and did not show any rescue of viable virus after temperature downshift and co‐cultivation with susceptible cells. Transformed cells maintained the transformed state in the absence of viral DNA. Our data therefore support the “hit‐and‐run” hypothesis for the transforming effect of HSV. © 1992 Wifey‐Liss, Inc.
Elevated stromelysin‐1 and reduced collagenase‐IV expression in invasive rat embryo fibroblasts expressing e1a deletion mutants + T24‐H‐ras
Authors: Engel Georg , Popowicz Przemyslaw , Marshall Heather , Norling Gertrud , Svensson Catharina , Auer Gert , Akusjärvi Göran , Linder Stig
Published: February 2007

We have studied the in vitro invasive properties of 3 cell lines derived from the co‐transfection of rat embryo fibroblasts (REF) with E1A genes deficient in exon 2 and T24‐ras. All 3 cell lines showed invasive properties at passage 10 after isolation. Invasive cells expressed elevated levels of stromelysin‐I and reduced levels of 68‐kDa type‐IV collagenase compared with untransfected REF. In 2 cell lines the invasive capacity increased during in vitro propagation. The expression of stromelysin‐I increased during this process, whereas 68‐kDa type‐IV collagenase was persistently expressed at reduced levels. In the third clone analyzed, the invasive capacity decreased during culture, in parallel with decreased expression of stromelysin‐I. The low level of stromelysin‐I expression observed in this cell line did not result from loss of AP‐I ‐transcription‐factor activity, and was not reversed by phorbol‐ester treatment. © 1992 Wifey‐Liss, Inc.
Fibroblast‐dependent tumorigenicity of melanoma xenografts in athymic mice
Authors: Gärtner Maria F. R. M. , Wilson E. Lynette , Dowdle Eugene B.
Published: February 2007

Two human melanoma cell lines, UCT‐Mel 2 and UCT‐Mel 3, were invariably tumorigenic in nude mice when inoculated s.c. in doses of 106 cells or higher; 105 cells or less did not give rise to tumours. In this report we show that otherwise sub‐tumorigenic inocula developed into vigorously growing tumour xenografts when co‐inoculated with normal fibroblasts. Fibroblasts derived from adult, neonatal or embryonic tissues all functioned as complementing cells, as did cells of human or murine origin. There was, however, a requirement for complementing cell viability, since ethanol‐killed fibroblasts were inefficacious. The fibroblast effect was dose‐dependent and was not observed if injections of fibroblasts and melanoma cells were separated anatomically or temporally. We have shown, by titrating admixtures of melanoma cells and fibroblasts, that fibroblasts are, in quantitative terms, more efficacious than melanoma cells as complementing cells. The system we describe provides a useful model for the study of stromal‐cell regulation of tumour growth. © 1992 Wifey‐Liss, Inc.
Arrest in late G2 or prophase of cell cycle induced by 4,4‐(l,2‐ethanediyl) bis (1‐isobutoxycarbonyloxymethyl 2, 6‐piperazinedione) (MST‐16) in cultured l1210 cells
Authors: Liu Yun‐Peng , Araya Shin‐Ichi , Nakamura Toru
Published: February 2007

The effects of MST‐16, a new antitumor agent derived from bis (2, 6‐dioxopiperazine), on cell growth, cell‐cycle progression and DNA synthesis, alone and in combination with other antitumor agents, were investigated in murine leukemia L1210 cells in vitro. The drug showed dose‐dependent inhibition of cell growth, and this effect was cell‐cycle phase‐specific. Flow cytometric analysis indicated that the drug could retard‐arrest the cells in late G2 phase or prophase and that it did not affect the progression from G1 to G2 phase. In the presence of MST‐16, the change in 3H‐thymidine incorporation was proportional to the retardation‐arrest of the cells, suggesting that MST‐16 has no direct action on DNA synthesis itself. MST‐16 could continuously retard the cells which were arrested by etoposide (VP‐16); and vincristine (VCR) could block the progression of the cells arrested by MST‐16, but not vice versa. The addition of MST‐16 followed by VCR was more effective than simultaneous addition of the 2 drugs on inhibition of cell growth. These results will be useful in designing a reasonable regimen of MST‐16 chemotherapy for malignancies. © 1992 Wifey‐Liss, Inc.
Metastatic phenotype of murine tumor cells expressing different cooperating oncogenes
Authors: Virone Angela , Monier Roger , Zerial Aurelio , Lavelle François , Feunteun Jean
Published: February 2007

Four murine cellular tumor models expressing various combinations of oncogenes (SV40 large T and v‐Ha‐ras, SV40 large T and v‐src, SV40 large T and neu, adenovirus E1A and v‐Ha‐ras) induce sarcoma when they are inoculated s.c. into the DBA/2 syngenic mice. The metastatic patterns, distribution and fate of these tumor cells transplanted by two different routes into syngenic DBA/2 mice have been studied. All the tumor cell lines except E1A‐ras, induce massive overt artificial metastases principally in the lung after i.v. injection. In s.c. tumor‐bearing mice, a few resting cells colonize the lung as micrometastases. When removed from this tissue context and injected s.c. these cells regain their proliferative potential and grow as local tumors which again give rise to occult pulmonary micrometastases. © 1992 Wifey‐Liss, Inc.
Colon cancer in argentina. II: Risk from fibre, fat and nutrients
Authors: Iscovich J. M. , L'abbé K. A. , Castelleto R. , Calzona A. , Bernedo A. , Chopita N. A. , Jmelnitzsky A. C. , Kaldor J. , Howe G. R.
Published: February 2007

A case‐control study has been conducted to investigate the relationship between total energy intake, fibre and nutrients and colon cancer in Argentina. Cases are 110 newly diagnosed patients from 10 hospitals between 1985 and 1986. Two neighbourhood controls per case were matched on age, sex and place of residence. The intake of calories, fibre and nutrients was estimated from the information collected on food consumption during the 5‐year period up to 6 months prior to interview of subjects, based on a pilot study and standard portion sizes in Argentina. In conditional regression models, dietary fibre is highly protective (odds ratio (OR) = 0.07 per 19.02 g/day; 95% confidence interval (CI): 0.02 to 0.25) and total energy intake increases risk (RR = 1.82 per 1000 kcal/day; 95% CI: 1.20 to 2.77), each with adjustment for the other. Analysis of caloric components (fat, protein and carbohydrates) reveals that carbohydrates are the most important factor driving the total energy effect. Other nutrients make little apparent contribution to risk.
Clustering of discrete cell properties essential for tumorigenicity and metastasis. III. Dissociation of the properties in N‐ras‐transfected rsv‐SR‐transformed cells
Authors: Deichman G. I. , Topol L. Z. , Kluchareva T. E. , Matveeva V. A. , Zakamaldina T. A. , Uvarova E. N. , Tatosyan A. G.
Published: February 2007

We have previously shown that RSV‐SR‐transformed hamster cells acquire high resistance to H2O2, i.e. the cytotoxic product of activated macrophages (H2OR2) and that they begin to secrete PGE (PGEs), thus inactivating the CTA of NK cells. Among normal cells, the same phenotype is expressed in activated macrophages. In all our RSV‐transformed cells these 2 properties were jointly expressed and correlated with high tumorige‐nicity and experimental metastasizing of these cells. We now show that transfection of 3 RSV‐SR‐transformed cell strains with activated N‐ras leads either to complete inhibition of the H2OR2 + PGES phenotype in all clones of one strain, or to inhibition of PGES only in the majority of clones of 2 other strains. Unexpectedly, the complete or partial inhibition of this phenotype did not alter the high tumorigenicity of 2 strains of these cells, but lower tumorigenicity was evident in almost all clones of the third strain (as well as in some gene‐neo‐transfected clones of these strains). The loss of PGES made these cells susceptible to the CTA of NK cells, while the loss of H2OR2 did not alter their resistance to the CTA of macrophages. Expression of the H2OR2 + PGES phenotype was retained in all cloned variants of control, gene‐neo‐transfected cells. The possible relation of the N‐ras gene to regulation of src gene activities in RSV‐SR‐transformed cells is discussed.