bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of sialoadenectomy on medroxyprogesterone‐acetate‐induced mammary carcinogenesis in BALB/c mice. Correlation between histology and epidermal‐growth‐factor receptor content
Authors: Kordon Edith C. , Guerra Fabiana , Molinolo Alfredo A. , Elizalde Patricia , Charreau Eduardo H. , Pasqualini Christiane Dosne , Montecchia Fernanda , Pazos Patricia , Dran Graciela , Lanari Claudia
Published: February 2007

To evaluate the possible involvement of the salivary glands in the modulation of medroxyprogesterone (MPA)‐induced mammary tumorigenesis, 48 sialoadenectomized virgin BALB/c female mice and 47 controls were treated with 40mg MPA depot s.c. every 3 months for 1 year. Mammary tumors developed in 11 sialoadenectomized and in 34 control mice with similar latencies. In both groups, 75% of the tumors were ductal and progestin‐dependent (PD) while the remainder were lobular and progestin‐independent (PI). Epidermal growth factor (EGF) levels were measured in salivary glands (SG‐EGF) and serum (S‐EGF) in both groups. MPA induced a significant increase in SG‐EGF and in S‐EGF that became evident only after 1 month of MPA treatment. No increase in S‐EGF was detected in MPA‐treated sialoadenectomized mice, indicating that salivary glands are the major source of S‐EGF. The presence of EGF receptors (EGF‐R) was investigated in ductal PD and PI tumor lines and compared with 8 PI tumor lines of lobular origin. A significant difference in EGF‐R content was found between lobular and ductal tumors. No increase in EGF‐R was noted when ductal tumors became autonomous. EGF‐R did not correlate with tumor growth rate and there was an inverse correlation between EGF‐R and steroid receptors. When the effect of sialoadenectomy on tumor growth was tested in vivo in syngeneic transplants of 2 ductal PD, I ductal PI and 2 lobular PI mammary adenocarcinomas, it was not found to be significant when compared with the controls. It may be concluded that SG‐EGF plays an important role in the induction of mammary adenocarcinomas by MPA, while it has no significant effect on the growth of established tumors.
DT‐diaphorase as a determinant of sensitivity to adriamycin in non‐small‐cell lung‐cancer cell lines
Authors: Kasahara Kazuo , Shibata Kazuhiko , Bando Takuma , Numata Yuka , Fujimura Masaki , Matsuda Tamotsu
Published: February 2007

We have reported the establishment of a mitomycin‐C (MMC)‐resistant non‐small‐cell lung‐cancer cell line, PC‐9/MC4. As determined by an MTT assay, this resistant cell line was found to be 4 times more sensitive to adriamycin (ADM) than was the parental PC‐9. There were no significant differences in sensitivity to etoposide, mitoxantrone, daunomycin, epirubicin, pirarubicin, 9‐aminoanthracycline or 3'‐deamino‐3'‐morpholino‐13‐deoxo‐10‐hydroxy carminomycin. These data suggest that neither qualitative or quantitative changes in DNA topoisomerase 11 nor the enhanced repair of DNA can explain the differing sensitivity to ADM observed. No significant differences were found in the accumulation of ADM and glutathione (GSH) in these cell lines. Although total glutathione‐S‐transferase (GST) activity in PC‐9/MC4 cells was lower than that observed in PC‐9 cells and treatment with ethacrynic acid (EA) reduced sensitivity to ADM in both cell lines, relative resistance was unaffected. NADH‐cytochrome b5 reductase (B5R) activity in PC‐9/MC4 cells showed a 3‐fold greater decrease than that in PC‐9 cells, and DT‐diaphorase (DTD) activity in PC‐9/MC4 cells showed an approximately 200‐fold greater decrease than that in PC‐9 cells. Addition of dicumarol, an inhibitor of DTD, decreased the sensitivity of ADM of PC‐9 but not of PC‐9/MC4. DTD activity in the PC‐9 cell line was inhibited by treatment with dicumarol while in PC‐9/MC4 it remained unchanged. These data suggest that DT‐diaphorase is a determinant of sensitivity to ADM in the 2 cell lines.
Induction of multidrug resistance (MDR) by transfection of MCF‐10A cell line with c‐Ha‐ras and c‐erbB‐2 oncogenes
Authors: Sabbatini Antonietta R. M. , Basolo Fulvio , Valentini Paola , Mattii Letizia , Calvo Simonetta , Fiore Lisa , Ciardiello Fortunato , Petrini Mario
Published: February 2007

To investigate the relationship between oncogene activation and appearance of multidrug resistance (MDR) we transfected the human breast epithelial cell line MCF‐10A, negative for the expression of the P‐glycoprotein, with c‐Ha‐ras and/or c‐erbB‐2 oncogenes. The appearance of the MDR phenotype was then studied by evaluating mdr‐1 mRNA expression, the presence of P‐glycoprotein on the cell membrane and the onset of doxorubicin resistance, together with the effect of the reversing agent verapamil. We found that only MCF‐10A transfected with both c‐Ha‐ras and c‐erbB‐2 oncogenes acquired the MDR phenotype.
Daunorubicin efflux against a concentration gradient in non‐P‐glycoprotein multidrug‐resistant lung‐cancer cells
Authors: Mülder H. Sipko , Lankelma Jan , Dekker Henk , Broxterman Henk J. , Pinedo Herbert M.
Published: February 2007

Multidrug‐resistant, human non‐small‐cell lung carcinoma SW‐I573/2RI20 (2RI20) cells, not containing the drug efflux pump P‐glycoprotein (Pgp), have reduced initial daunorubicin (DN) accumulation rates and decreased cellular steady‐state drug concentrations. Previously we found indications of the presence of a plasma membrane “vacuum cleaner”, pumping DN directly from the membrane, and reported evidence of active DN pumping using digitonin. Further evidence of active DN pumping is now provided via a different methodology and the active drug pump flux is estimated. Cells were exposed to a flowing medium containing the cytotoxic agent DN. After reaching a steady state, in which net DN uptake equals net DN efflux, high concentration pulses of vincristine (VCR) were injected into the flowing medium. A rapid increase in cellular DN content was observed, while only a minimal effect was seen in SW‐I573 wild‐type cells. After passage of the VCR pulse, the extra accumulated DN was effluxed against a concentration gradient. Upon increasing the VCR concentration, a maximum pump inhibition was reached which was similar to the effect of cellular energy depletion. Similar effects were observed for Pgp‐containing SW‐I573/2RI60 (2RI60) cells as well as non‐Pgp MDR human small‐cell lung carcinoma GLC4/ADR cells. With increasing extracellular DN concentrations, saturation of the VCR‐induced DN influx was observed (DN medium concentration 2.5 μM at 1/2 Vmax). At an extracellular DN concentration of 5 μM, higher concentrations of VCR were needed to reach the maximum effect in 2RI20 cells than at 0.5 μM DN. This is an indication of competitive interaction between DN and VCR for the putative DN efflux system. In summary, we found indications of inhibition of active DN efflux by VCR and DN efflux against a concentration gradient in non‐Pgp MDR 2RI20 and GLC4/ADR cells. These features are consistent with the presence of a multidrug transporter, different from Pgp, in the plasma membrane of these cells.
Expression of MUC2‐mucin in colorectal adenomas and carcinomas of different histological types
Authors: Blank Monika , Klussmann Enno , Krüger‐Krasagakes Sabine , Schmitt‐Gräff Anette , Stolte Manfred , Bornhoeft Gudrun , Stein Harald , Xing Pei‐Xiang , McKenzie Ian F. C. , Verstijnen Cees P. H. J. , Riecken Ernst Otto , Hanski Christoph
Published: February 2007

The expression of mucin MUC2 was investigated in normal colonic tissue, in colonic adenomas and in carcinomas of the mucinous and non‐mucinous type. The latter were subdivided into carcinomas originating from the adenoma‐carcinoma sequence (ACS) and de novo (DN) carcinomas. The expression was assayed by immunohistochemistry with the monoclonal anti‐MUC2 antibody CCP58 and by mRNA semiquantitation. MUC2 protein epitope CCP58 was strongly expressed in 21 % of normal colonic tissues, in 40% of villous and in 48% of tubular adenomas. Mucinous carcinomas exhibited strong expression in 72%, ACS carcinomas in 21 % and DN adenocarcinomas in none of the tumors investigated. Compared with the adjacent non malignant tissue (transitional mucosa), CCP58 epitope expression in the tumor was higher in 74% of mucinous carcinomas, but equal or lower in 69% of ACS carcinomas and in 100% of de novo carcinomas. The alterations of MUC2 expression detected by immunohistochemistry in adenocarcinomas were confirmed on mRNA level. These data indicate that the MUC2 expression pattern is different in the 3 carcinoma types investigated. MUC2 over‐expression occurs in the adenomatous tissue. It is always maintained in mucinous carcinomas, but frequently decreased in non‐mucinous ACS carcinomas. DN carcinomas are most frequently associated with decreased expression of MUC2. © 1994 Wiley‐Liss, Inc.
Human papillomavirus DNA in cervical carcinoma—correlation with clinical data and influence on prognosis
Authors: Ikenberg Hans , Sauerbrei Willi , Schottmüller Ursula , Spitz Christian , Pfleiderer Albrecht
Published: February 2007

Human papillomavirus (HPV) is a main factor in cervical carcinogenesis. However, data on the correlation of HPV with clinical features and the prognosis of cervical carcinoma remain controversial. The HPV status (positivity, type, copy number) in unfixed tissue specimens of 205 primary invasive cervical carcinomas was determined by Southern blot hybridization. A correlation with comprehensive clinical and histopathologic data and long‐time survival was evaluated. HPV DNA was detected in 73% of the cases; 83% of the HPV‐positive tumors contained HPV 16. HPV 16 was predominant among squamous‐cell carcinomas (SCC) (ρ = 0.05). HPV 16 copy number was higher in keratinizing tumors (ρ < 0.05), and elevated levels of the SCC antigen were more common in patients positive for HPV 16 (ρ < 0.03). No association was found between the HPV status and 8 other clinical and histopathologic variables. Multivariate analysis after a median follow‐up of 73 months demonstrated longer survival for patients with lower clinical stage (ρ = 0.001) and keratinizing SCC (ρ = 0.005). Women with HPV‐negative tumors had a higher risk of death (RR 1.51; ρ = 0.07). HPV analysis does not clearly define biologically distinct sub‐sets of cervical carcinoma. This underlines the importance of additional factors in cervical carcinogenesis. © 1994 Wiley‐Liss, Inc.
Urbanization gradients and cancer mortality in Uruguay, 1988–1992
Authors: Vassallo Alberto , De Stéfani Eduardo , Ronco Alvaro , Barrios Enrique
Published: February 2007

Cancer deaths occurring in Uruguay in the period 1988–1992 were classified by site, sex and residence according to the ninth version of the International Classification of Diseases (ICD‐9), in order to analyze gradients of urbanization. For this purpose, Uruguayan counties were classified as rural, suburban and urban, using population density as an ordinal indicator of the degree of urbanization. Significant increasing trends with urban excess were observed for cancers of the colon, rectum, lung and female breast. In addition, this study showed significant urban deficits for esophageal, gastric and uterine cancers, contrasting with previous studies. These differences seem to be real rather than artifactual due to misclassification. The study thus provides interesting new findings concerning urban differences between developed communities and a partially developing country. © 1994 Wiley‐Liss, Inc.
Repair of cisplatin‐DNA adducts by protein extracts from human ovarian carcinoma
Authors: Jones Sandra L. , Hickson Ian D. , Harris Adrian L. , Harnett Paul R.
Published: February 2007

Human ovarian carcinoma exhibits a spectrum of responses to treatment with cisplatin, ranging from marked sensitivity to relative resistance. Variations in the efficiency with which cells repair cisplatin‐DNA adducts may in part determine cisplatin sensitivity. We have investigated the use of a novel cell‐free assay of nucleotide excision repair of platinated DNA to assess the DNA repair capacity of direct biopsies of untreated human ovarian carcinoma. In this pilot study, ovarian carcinoma extracts performed in vitro excision repair of platinated DNA. Moreover, tumors from different individuals varied as much as I O‐fold in their repair capacity in this assay. © 1994 Wiley‐Liss, Inc.
Detection of neutralizing antibodies against human T‐cell leukemia virus type 1 using a cell‐free infection system and polymerase chain reaction
Authors: Haraguchi Yuji , Yang Da‐Wei , Handa Atsushi , Shimizu Nobuaki , Tanaka Yuetsu , Hoshino Hiroo
Published: February 2007

We have developed a cell‐free infection system to titrate neutralizing antibodies against human T‐cell leukemia virus type I (HTLV‐I) using the polymerase chain reaction (PCR). S+L‐CCC (8C) feline kidney or U‐251 MG human glioma cells were infected with a cell‐free culture supernatant derived from HTLV‐I ‐infected c77 feline cells. DNA was extracted from 8C or U‐251 MG cells after incubation for 24 hr and amplified by PCR. The c77 cell supernatant gave discrete bands, whereas those of HTLV‐I ‐positive T cells did not. When the inocula were treated with HTLV‐I antibody‐positive human sera or the monoclonal or polyclonal antibody against the peptide 190–199 of HTLV‐I envelope protein gp46, the subsequent formation of HTLV‐I proviral DNA was inhibited. We determined the titers of neutralizing antibodies by densitometrically scanning the intensity of the PCR bands. These titers correlated well with those determined by the plaque assay using a pseudotype of vesicular stomatitis virus bearing the envelope antigens of HTLV‐I. At high serum concentrations, many seronegative samples markedly inhibited the plating of the HTLV‐I pseudo‐type whereas they barely affected results obtained by PCR. Thus, the c77‐PCR system can detect neutralizing antibodies against HTLV‐I even at low titers. © 1994 Wiley‐Liss, Inc.
Childhood cancer incidence in relation to distance from the former nuclear testing site in semiplatnsk, kazakhstan
Authors: Zardze Daid G. , Li N. , Men Tamara , Duffy Spephen W.
Published: February 2007

Rates of childhood cancer between 1981 and 1990 in the 4 administrative zones of Kazakhstan were studied to asess the relationship, if any, with distance from nuclear testing sites. Risk of various cancers among children aged 14 years or younger were estimated in relation to distance from (1) a site where testing in air was performed before 1963, (2) a site where underground testing took place thereafter, and (3) a reservoir, known as “Atom Lake,” created by 4 nuclear explosions in 1965. Risk of acute leukaemia rose significantly with increasing proximity of residence to the testing areas, although the absolute value of the risk gradient was relatively small. The relative risk for those living less than 200 km from the airtesting site was 1.76 compared with those living 400 km or more away from the site. Similar relative risks were observed for the underground site and “Atom Lake.” There was also some evidence of increased risk of brain tumours in association with proximity to the test sites. In 2 of the 4 zones studied, there was substantial regional variation in acute leukaemia rates which was not attributable to distance from the test site. The findings may be affected by potential confounders, notably urban/rural status and ethnic factors. © 1994 Wiley‐Liss, Inc.
A case‐control study of reproductive risk factors associated with cervical cancer
Authors: Mukherjee Bishwa Nath , Sengupta Subrata , Chaudhuri Salil , Biswas Liton Naha , Maiti Pradip
Published: February 2007

Using the logistic‐regression technique, a hospital‐based case‐control study of 177 married women with invasive squamous‐cell cervical cancer and 149 hospital‐visiting controls enabled evaluation of selected reproductive factors as risks. Early age at marriage was found to be the single best predictor of the disease status. However, those who married late but gave birth to a large number of children were generally found to be suffering from cervical cancer. The results support the hypothesis that it is not so much parity per se that enhances the risk, but the rapidity of multiple pregnancies that matters. Logistic analysis also revealed the independent influence of birth interval on the risk of cervical cancer. These findings warrant serious consideration in future studies, given the obvious implications for prevention. Other implications for the prevention of cervical cancer are briefly discussed. © 1994 Wiley‐Liss, Inc.
Asbestos and erionite fibres can induce mutations in human lymphocytes that result in loss of heterozygosity
Authors: Both Katrin , Henderson Douglas W. , Turner David R.
Published: February 2007

Although asbestos and erionite are proven human carcinogens, most studies have concluded that these fibres are not mutagenic to mammalian cells in vitro. We have studied the potential of these fibres and chrysotile fibres to induce mutations in human peripheral lymphocytes, using a mutation assay that measures mutation at the autosomal HLA‐A locus. Exposure of lymphocytes in culture to 400 μg/ml of crocidolite or erionite for 72 hr did not result in a statistically significant increase in the mutation frequency (MF) in the HLA‐A assay, although a trend towards increased MF was observed. Exposure to 400 μg/ml chrysotile resulted in no increase in MF; however a significant increase was observed at 50 μg/ml. Mutations in somatic cells can be classified according to their molecular basis. Molecular analysis of mutants obtained following exposure of lymphocytes to crocidolite and erionite demonstrated a statistically significant increase in the class of mutations arising from loss‐of‐heterozygosity (LOH) events involving the selection locus (HLA‐A) and more distal loci. Mutations following exposure to crocidolite and erionite showed a greater frequency of LOH than did spontaneous mutants (p < 0.02 and p < 0.005 respectively). Mutants following exposure to chrysotile did not display a significant difference in LOH when compared with spontaneous mutants. Thus, although an increase in overall mutation frequency following fibre exposure did not achieve statistical significance, the modest increase seen following exposure to erionite and crocidolite is translated into a highly significant change in those components of the spectrum of mutations which result in LOH. © 1994 Wiley‐Liss, Inc.
Hepatic tumors induced by carbon tetrachloride in transgenic mice carrying a human c‐H‐ras proto‐oncogene without mutations
Authors: Tsunematsu Satoshi , Saito Hidetsugu , Kagawa Tatehiro , Morizane Toshio , Hata Jun‐Ichi , Nakamura Tatsuya , Ishii Hiromasa , Tsuchiya Masaharu , Nomura Tatsuji , Katsuki Motoya
Published: February 2007

Hepatic tumors were generated in mice by repeated administration of carbon tetrachloride (CCI4). Eight transgenic (Tg) mice carrying a human c‐H‐ras proto‐oncogene (rasH2 line) and 9 non‐Tg mice were killed at 20 weeks. Tg mice developed more tumors than did non‐Tg littermates. Most tumors were neoplastic nodules, but I hepatocellular carcinoma (HCC) was found in a Tg mouse at 20 weeks. Three Tg and 2 non‐Tg mice were kept without further administration of CCI4. Two Tg mice died at 30 weeks of HCC with intra‐abdominal bleeding, and I Tg mouse developed HCC with a mesenteric metastasis at 32 weeks. No HCC was found in 2 non‐Tg mice at 32 weeks. Although mutations at codon 12, 13, and 61 of the H‐ras gene are often found in murine hepatocarcinogenesis, neither the tumors, including one HCC, nor the normal cells revealed any such mutations. These results showed that the unmutated human c‐H‐ras gene facilitates malignant transformation of hepatocytes when continuous liver‐cell death and regeneration is caused by repeated administration of CCI4. © 1994 Wiley‐Liss, Inc.
Correlation between cytopathological results and In situ hybridisation on needle aspiration biopsies of suspected african Burkitt's lymphomas
Authors: Labrecque Louise G. , Lampert Irvin , Kazembe Peter , Philips John , Griffin Beverly E.
Published: February 2007

Burkitt's lymphoma (BL) is a very high‐incidence malignancy in sub‐Saharan Africa, where it targets mainly young children. This lymphoma is closely associated with Epstein‐Barr virus (EBV). Diagnosis of BL relies on clinical presentation as well as histological results obtained from biopsies. In this report, 66 new patients from Malawi (one of the southernmost African countries with high‐incidence BL), suspected on clinical grounds to present with BL, had fine needle aspiration biopsies taken, smeared on slides and used for May‐Grünwald Giemsa staining. Duplicate slides were independently assessed for EBV presence and expression by DNA‐DNA and/or RNA‐RNA in situ hybridisation (ISH), using respectively the repetitive viral BamHIW DNA fragment in a biotinylated probe and the small EBV‐encoded RNA EBER I in a digoxigenin‐labelled riboprobe. There was very good correlation between the various techniques in the diagnosis of the lymphomas, showing 67% of clinically suspect cases to be BL. Our report, presenting data on BL in Malawi, illustrates the usefulness of a simple aspiration biopsy in the diagnosis of this malignancy by Giemsa staining and also in both types of ISH.
Frequent loss of heterozygosity of the long arm of chromosome 7 is closely associated with progression of human gastric carcinomas
Authors: Kuniyasu Hiroki , Yasui Wataru , Yokozaki Hiroshi , Akagi Morihisa , Akama Yoshihiko , Kitahara Kenji , Fujii Kiyomu , Tahara Eiichi
Published: February 2007

Loss of heterozygosity (LOH) on the long arm of chromosome 7 was examined using 5 polymorphic marker probes on 98 gastric carcinomas to elucidate a novel locus for development and progression of the tumors. Twenty‐six (32%) of 82 informative cases showed LOH on 7q on at least one locus of 5 loci. Among 5 loci, LOH at D7S95 locus was most frequent, the incidence being 53% in well‐differentiated gastric carcinomas and 33% in poorly differentiated and scirrhous gastric carcinomas respectively. At 3 loci, c‐met, D7S63 and D7S22, the incidence of LOH was about 30% and 10% in well‐differentiated and poorly differentiated gastric carcinoma cases respectively. In contrast, LOH at D7S64 was not detected in any gastric‐carcinoma cases. Deletion mapping of 7q revealed that D7S95 locus was the essential region of LOH. Eight (62%) of 13 cases with LOH at D7S95 locus belonged to the most advanced stage grouping. Furthermore, 6 (75%) of 8 cases with abdominal dissemination showed LOH at D7S95. Therefore, cases with LOH at D7S95 showed significantly worse prognosis than the cases without the LOH in the stage‐III and stage‐IV groups. These findings overall suggest that D7S95 locus on 7q may contain a candidate suppressor gene for the progression of gastric carcinoma.
Topoisomerase I and II activity in human breast, cervix, lung and colon cancer
Authors: McLeod Howard L. , Douglas Fiona , Oates Melanie , Symonds R. Paul , Prakash Drew , Van Der Zee Ate G. J. , Kaye Stanley B. , Brown Robert , Keith W. Nicol
Published: February 2007

The identification of human DNA topoisomerases as cellular targets for active anti‐cancer drugs has stimulated further interest in topoisomerase function in tumour biology. Topoisom‐erase I and II catalytic activity is detectable in many normal and malignant tissues. However, little is known about the expression of topoisomerases in most human solid tumours. The present study evaluated topoisomerase I and II activity in biopsy samples from 86 patients with breast, lung, cervix or colon cancers. Significant intra‐ and inter‐tumour variation in topoisomerase expression was observed. Topoisomerase I activity was relatively high in cervix and colon tumours in comparison to lung and breast cancers. Topoisomerase II activity was high in cervix, colon and lung cancers relative to breast cancer. Topoisomerase I and II activity co‐segregated in individual colon tumour samples, but no correlation was observed in cervix, lung or breast tumours. The large heterogeneity in both topoisomerase I and II activity within a tumour type suggests a mechanism for variable response to topoisomerase‐directed therapy. The differences in activity between tumour groups suggest that the potential efficacy of inhibitors of topoisomerase I in colon and cervical tumours may be greater than in lung and breast tumours. Future in vivo evaluation is required to establish the clinical relevance of the observed heterogeneity in topoisomerase activity.
Parallel assessment of glutathione‐based detoxifying enzymes, O6‐alkylguanine‐dna alkyltransferase and P‐glycoprotein as indicators of drug resistance in tumor and normal lung of patients with lung cancer
Authors: Oberli‐Schrämml Aurelia E. , Joncourt Franziska , Stadler Markus , Altermatt Hans‐Jorg , Buser Kathrin , Ris Hans‐Beat , Schmid Ulrico , Cerny Thomas
Published: February 2007

The levels of several potential indicators of drug resistance were measured in tumor and corresponding normal tissue of 55 untreated patients with lung cancer. The resistance parameters include glutathione (GSH) level, activities of the enzymes glutathione transferase (GST), glutathione peroxidase (GPx) and O6‐alkylguanine‐DNA alkyltransferase (ATase), as well as expression of P‐glycoprotein (Pgp). Median values of GSH, GST and GPx were significantly higher in tumor than in normal tissue of non‐small‐cell lung cancer (NSCLC) or of small‐cell lung cancer (SCLC), whereas ATase was elevated in tumor tissue of NSCLC only. Pgp expression as determined by Western blotting was significantly lower in tumor than in normal tissue of NSCLC. Resistance‐parameter expression did not correlate with stage of disease or age of the patients. We found a negative correlation between smoking intensity and GSH level in normal tissue. Our findings indicate that the fundamental differences in chemo‐sensitivity between SCLC and NSCLC cannot be explained by differences in the GSH‐system or in the expression of Pgp. However, the level of ATase activity may be one of the factors responsible for the difference in chemosensitivity.
Presence of immunoassayable transforming growth factor‐β1 (tgf‐β1) in breast cyst fluid (BCF): Relationship with the intracystic electrolyte and epidermal‐growth‐factor (EGF) content
Authors: Zanard S. , Pensa F. , Torrisi R. , De Franchis V. , Barreca A. , Minuto F. , Boccardo F.
Published: February 2007

We evaluated the presence and distribution of transforming growth factor‐β1 (TGF‐β1) in breast cyst fluid (BCF), and its relationship with intracystic epidermal growth factor (EGF). EGF and TGF‐β1 were determined by radioimmunoassay on 47 BCFs (27 of the Na+/K+ <3 type and 20 of the Na+/K+ >3 type). As expected, EGF levels were inversely correlated with the Na+/K+ ratio, and were consequently higher in Na+/K+ <3 cysts as compared with Na+/K+ >3 cysts, (p < 0.005). By contrast, TGF‐β1 levels were directly correlated with the Na+/K+ ratio (p < 0.01), being higher in Na+/K+ >3 cysts, though not significantly (p = 0.057). A significant negative relationship was found between EGF and TGF‐β1 concentration. When the analysis was performed separately in the 2 cyst sub‐populations, EGF and TGF‐β1 were found to be negatively and significantly correlated in Na+/K+ <3 cysts only (p < 0.01). Our results demonstrate that Na+/K+ <3 cysts contain high levels of EGF, a growth‐stimulating factor, and very low levels of TGF‐β1, a growth‐inhibiting factor. This may provide an explanation for the higher risk of breast cancer observed in women with Na+/K+ <3 cysts. Our results also suggest that EGF accumulation in this type of cysts might be regulated by TGF‐β1.
Enzyme‐Linked immunosorbent assay (ELISA) for IgA and IgG antibodies to epstein‐barr‐virus ribonucleotide reductase in patients with nasopharyngeal carcinoma
Authors: Fones‐Tan A. , Chan S. H. , Tsao S. Y. , Gan L. H. , Tan W. H. , Li B. , Khong P. W. , Gan Y. Y.
Published: February 2007

661 bp coding for the carboxyl end of the large sub‐unit of EBV ribonucleotide reductase was cloned into the pMal plasmid vector. Purified recombinant protein was tested in IgG and IgA ELISAs. For the IgG assay, 81 out of 100 NPC patients tested positive, whereas for the IgA assay, 60 tested positive. Among 100 normal individuals, I tested positive for the IgG assay and 9 tested positive for the IgA assay. The IgG assay picked up 6 out of 19 NPC sera which were IFA‐VCA‐ and IFA‐EA‐negative for IgA antibodies. Hence the recombinant ribonucleotide reductase could have good potential as a diagnostic test for NPC or could serve as a complementary test to IFA.
The decreased expression of CD3 ζ chains in cancer patients is not reversed BY IL‐2 Administration
Authors: Farace Françoise , Angevin Eric , Vanderplancke Josiane , Escudier Bernard , Triebel Frédéric
Published: February 2007

The mechanisms underlying the impaired immune response frequently observed in cancer patients are poorly understood. Recently, a decreased expression of the signal‐transducing CD3 ζ chains has been shown to occur in patients with colorectal or renal‐cell carcinoma. Here, we have investigated the expression of the ζ molecule on lymphocytes from patients with advanced cancers including renal‐cell carcinoma (n = 21), hepatic colorectal carcinoma metastases (n = 6), head‐and‐neck squamouscell carcinoma (n = 7) or miscellaneous tumors (n = 7), using flow cytometry analysis of PBMCs with monoclonal antibody specific for the cytoplasmic domain of the ζ chain. The same analysis was also performed with patients undergoing IL‐2 therapy (from 5 days up to 3 months i.v. or s.c). PBMCs from patients (n = 41) showed a significant decrease in expression of the ζ chains as compared to the levels found in lymphocytes from healthy controls (n = 15), which was not reversed by IL‐2 administration. Together, these results suggest that an alteration in the expression of the ζ‐chain‐transducing element in PBMCs may be a common cause of progressive immunosuppression in advanced cancer patients and that IL‐2 is not sufficient to reverse this situation in vivo.
Menstrual and reproductive factors and Gastric‐cancer risk in women
Authors: La Vecchia Carlo , D'Avanzo Barbara , Franceschi Silvia , Negri Eva , Parazzini Fabio , Decarli Adriano
Published: February 2007

To understand possible correlates of the systematically higher gastric‐cancer rates in males than in females, we investigated the role of menstrual, reproductive and hormonal factors in females, using data from a case‐control study conducted in Northern Italy. Cases were 229 post‐menopausal women with incident, histologically confirmed gastric cancer, and controls were 614 post‐menopausal women in hospital for acute, nonneoplastic, non‐digestive‐tract conditions. After allowance for age, education, family history, and selected dietary correlates of stomach cancer, a reduced gastric‐cancer risk was observed in women with later menopause (odds ratio, OR = 0.6, p value for trend <0.05), and longer duration of fertile life (OR = 0.7, but the trend in risk was not significant). The risk of gastric cancer was elevated in multiparous women (OR = 1.7 for 3 and 1.9 for ≥ 4 births as compared to nulliparae). No association was observed with age at menarche, age at first birth, and with number of spontaneous or induced abortions. Ever‐users of oral contraceptives had a non‐significant increased risk (OR = 1.3), and users of oestrogen‐replacement treatment a non‐significant reduced risk (OR = 0.5). Although the association with each factor was moderate, and the interpretation of these findings is not straightforward, these results are compatible with the hypothesis that some role is played by (endogenous and exogenous) female hormones in the process of gastric carcinogenesis.
Linkage analysis in familial melanoma kindreds to markers on chromosome 6p
Authors: Walker Graeme J. , Nancarrow Derek J. , Walters Marilyn K. , Palmer Jane M. , Weber James L. , Hayward Nicholas K.
Published: February 2007

Malignant melanoma occurs as a familial cancer in 5%‐10% of cases, where it segregates in a manner consistent with autosomal dominant inheritance. Evidence from cytogenetics, fine mapping studies of deletions in melanomas and recent linkage studies supports the location of a human melanoma predisposition gene on the short arm of chromosome 9. Evidence also exists for a melanoma gene on I p, indicating genetic heterogeneity for melanoma predisposition. Previous studies have also reported findings suggestive of linkage of some melanoma families to the HLA region on the short arm of chromosome 6 (6p), indicating the possibility of even greater heterogeneity. To further define the possible effect of a gene within the HLA region on melanoma susceptibility, we have typed 7 simple tandem repeat polymorphisms (STRPs) from 6p in 16 Australian melanoma kindreds. Maximum 2‐point LOD scores ranged from 1.13 (θ = 0.2) to 2.03 (θ = 0.15) for 4 contiguous markers flanking the HLA complex, and multi‐point analysis gave a peak LOD score of 1.64, 24 centimorgans telomeric to D6S109. However, extended haplotype analysis of these markers showed that a region between D6S105 and HLAF segregated with melanoma in 5/16 families. These results are surprising given that the same cohort of families has previously been shown to be linked to chromosome 9. One interpretation of the current findings is that melanoma susceptibility in some families may result from a gene mapping within the HLA region of chromosome 6p.
P‐glycoprotein expression in the Golgi apparatus of multidrug‐resistant cells
Authors: Molinari Agnese , Cianfriglia Maurizio , Meschini Stefania , Calcabrini Annarica , Arancia Giuseppe
Published: February 2007

The surface and intracellular expression of mdrl‐P‐glycoprotein in parental drug‐sensitive human breast cancer cells (MCF‐7) and their multidrug‐resistant (MDR) variants has been studied by using the monoclonal antibodies (MAbs) MM4.17 and MRK‐16, which recognize 2 different epitopes of the drug efflux pump molecule. Fluorescence microscopic observations showed that P‐glycoprotein, in addition to being located at the cell surface, can also be found in the Golgi apparatus of resistant cells. To confirm this finding, Golgi apparatus and P‐glycoprotein were double‐labelled with wheat‐germ agglutinin (WGA) and MAb MM4.17. Laser scanning confocal microscopy indicated that, in MDR cells, Adriamycin mainly accumulated cytoplasmically in a perinuclear region. This accumulation proved to be modulated by pre‐treatment with verapamil or ATP depletion. Moreover, the vital staining of Adriamycin‐treated MDR cells, performed with the fluorescent lipid N‐{7‐(4‐nitrobenzo‐2‐oxa‐1,3‐diazole)}‐6‐aminocaproyl sphingosine (C6‐NBD‐ceramide), revealed that the anthracyclinic antibiotic was located in the Golgi apparatus. All these results indicate that the drug transporter is located in the Golgi apparatus, in which Adriamycin molecules also accumulate.
A secreted mucin carrying Sialyl‐Lewis a from colon carcinoma cells binds to E‐selectin and inhibits HL‐60 cell adhesion
Authors: Zhang Ke , Baeckström Dan , Hansson Gunnar C.
Published: February 2007

Sialyl‐Lewis × and a are known as ligands for E‐selectin (ELAM‐I) involved in leukocyte‐endothelial adhesion. L‐Canag (light cancer antigen), secreted by a colon carcinoma cell line COLO 205, is a soluble mucin‐type glycoprotein expressing sialyl‐Lewis a antigens. L‐Canag was purified from spent culture medium by trichloracetic acid precipitation and Superose 6 gel filtration. With a monoclonal antibody against E‐selectin (BBAI) as a positive control, the purified L‐Canag was shown to bind to E‐selectin‐Fc coated into plastic microtiter wells and to the surface of transiently E‐selectin‐transfected COS‐I cells in a Ca2+‐dependent way. Immunofluorescent double labelling showed that both BBAI and L‐Canag stained the same cells and morphological co‐localization on E‐selectin‐transfected COS‐I cells. Like BBAI, L‐Canag can inhibit leukocyte HL‐60 cell adhesion to E‐selectin‐transfected COS‐I cells, and this inhibition can be blocked by a F(ab')2 fragment directed against the sialyl‐Lewis a epitope.
Cellular immune response to human renal‐cell carcinomas: Definition of a common antigen recognized by HLA‐A2‐restricted cytotoxic T‐Lymphocyte (CTL) clones
Authors: Bernhard Helga , Karbach Julia , Wölfel Thomas , Busch Petra , Störkel Stefan , Stöckle Michael , Wölfel Catherine , Seliger Barbara , Huber Christoph , Zum Büschenfelde Karl‐Hermann Meyer , Knuth Alexander
Published: February 2007

Cytotoxic T lymphocyte (CTL) clones directed against autologous renal‐cell carcinoma (RCC) cell lines were generated by mixed lymphocyte/tumor‐cell culture (MLTC) using peripheral blood lymphocytes (PBL). A CD8+, CD4‐ CTL clone MZ1257‐CTL 5/30 with high cytolytic activity for the autologous tumor cell line MZ1257‐RCC was established. No lysis of the autologous EBV‐transformed B lymphocytes (EBV‐B) or K562 cells was observed. A panel of HLA‐A2‐matched allogeneic RCC lines was recognized by CTL 5/30. Further specificity analysis showed a cross‐reactivity with HLA‐A2‐matched allogeneic tumor cells of various origins, especially melanoma. CTL 5/30 was also cross‐reactive with several HLA‐A2‐positive allogeneic normal kidney cells in culture. The restriction element identified for CTL 5/30 was HLA‐A2, as shown by blocking of cytotoxicity using an anti‐HLA‐A2 monoclonal antibody (MAb) and by resistance of an HLA‐A2‐negative melanoma variant SK29‐MEL. 1.22 against lysis by CTL 5/30. In this report we demonstrate HLA‐A2‐restricted recognition of a T‐cell‐defined antigen on autologous renal‐cancer cells. This antigen is also expressed and recognized in association with HLA‐A2 on normal kidney cells in culture and other HLA‐A2‐positive tumor cells. It may therefore be a normal differentiation antigen to which tolerance is incomplete in the renal‐cell cancer system investigated.
Chimeric transcripts with an open reading frame are generated as a result of translocation to the Pvt‐1 region in mouse B‐cell tumors
Authors: Huppi Konrad , Siwarski David
Published: February 2007

Some mouse plasmacytomas exhibit a t(6;15) chromosomal translocation in which the breakpoint resides within the Pvt‐1 locus located 260 kilobases (kb) downstream of c‐myc on mouse chromosome 15. In this report, we show that the Pvt‐1 locus does not exhibit allelic exclusion in that Pvt‐1 transcripts continue to be expressed from the non‐translocated allele in t(6;15) plasmacytomas. From the translocated allele, we find chimeric transcripts containing a short 57‐bp segment of Pvt‐1 (termed Pvt‐1a) spliced directly to the immunoglobulin constant region sequence (lg‐Ck). These short transcripts have replaced a Jk segment with a trytophan residue via RNA splicing and contain a continuous open reading frame (ORF) from Pvt‐1a through Ck. Since this Pvt‐1a/Ck transcript is found in all 3 t(6;15) plasmacytomas examined, regardless of the location of the chromosomal breakpoint, we suggest that the Pvt‐1a/Ck chimera may have a functional role in the development of mouse plasmacytomas.
Association of transforming growth factor alpha (TGFA) and its precursors with malignant change in Barrett's epithelium: Biological and clinical variables
Authors: Brito Maria J. , Filipe M. Isabel , Linehan Jacqueline , Jankowski Janusz
Published: February 2007

Adenocarcinomas of the gastro‐esophageal junction (GEJ) and those arising in Barrett's esophagus (BE) are increasing in the West and have a poorer prognosis than distal stomach cancers. This has been attributed mainly to anatomical location, but biological factors such as growth‐regulatory molecules have been implicated. We have investigated the expression of one of these factors, TGFα, and its precursor prepro TGFα in 82 adenocarcinomas of GEJ (32 resected specimens and 50 biopsies) as well as in 48 BE biopsies without tumor, by immunohis‐tochemistry and by Western‐blot analysis. TGFα staining was shown in the cytoplasm and membrane of cells. Western blot confirmed that most immunoreactivity was against mature TGFa (5.6 kDa), but higher‐molecular‐weight bands were also identifiable, suggesting some reactivity with prepro protein. TGFα expression was more extense and intense in intestinal metaplasia and cancer. The tubular histological type of adenocarcinoma was more often positive than the signet‐ring type. Primary tumors with lymph‐node metastases also had increased TGFα expression. We conclude, therefore, that there is differential regulation of the expression of TGFα and its precursors during esophageal tumorigenesis. © 1995 Wiley‐Liss, Inc.
Prognosis following locally recurrent soft‐tissue sarcoma. A staging system based on primary and recurrent tumour characteristics
Authors: Choong Peter F. M. , Gustafson Pelle , Willén Helena , Åkerman Måns. , Baldetorp Bo , Fernö Mårten , Alvegård Thor , Rydholm Anders
Published: February 2007

We have shown that the clinical growth rate of local recurrence from soft‐tissue sarcoma could be expressed as a growthrate index (GRI) which was predictive for metastasis, and which was able to identify 2 equal populations of good (80% 2‐year MFS) and poor survivors (33%). We now report the associations between characteristics of the primary and GRI, and combine primary and locally recurrent tumour characteristics in a staging system. We studied 460 adult patients with soft‐tissue sarcomas of the extremities and trunk wall who were diagnosed and treated between 1964 and 1990, of whom 134 developed local recurrences and 151 metastases. The association of primary tumour size, histologic malignancy grade, depth, spontaneous necrosis, intratumoral vascular invasion and S‐phase fraction with local recurrence, GRI and metastasis were examined. High GRI was associated with primary tumours that were larger, deeper, more malignant, underwent spontaneous tumour necrosis, demonstrated intravascular invasion and had a higher S‐phase fraction. The same factors were also strongly associated with the incidence of metastasis. A multivariate analysis found GRI and primary tumour necrosis to be the strongest and most significant prognostic factors. GRI and tumour necrosis were combined in a staging system that identified groups with good survival (79 to 94% 2‐year MFS), intermediate survival (61% 2‐year MFS) and exceptionally poor survival (6% 2‐year MFS). These findings validate our earlier assertion that high GRI reflects highly malignant tumours. A staging system composed of primary tumour necrosis and GRI can identify patients who may be suitable candidates for trials of adjuvant chemotherapy. © 1995 Wiley‐Liss, Inc.
The expression profile of alternatively spliced neuronal c‐src RNA distinguishes between human tumours of the sympatho‐adrenal lineage
Authors: Örtoft Eva , Bjelfman Catarina , Hedborg Fredrik , Grimelius Lars , Påhlman Svcn
Published: February 2007

Human neuronal and neuroendocrine tumour specimens and cell lines were analysed regarding proteins and transcripts coded by the proto‐oncogene c‐src. At the protein level, most of the neuroblastomas and phaeochromocytomas expressed the neuronal c‐src form, pp60c‐srcN. None of the other neuroendocrine tumours, i.e. paragangliomas, neuroendocrine pancreatic tumours, or carcinoid tumours and small‐cell lung carcinomas of different types, appeared to express the neuronal form. In the brain, c‐src is transcribed into 3 differently spliced mRNA variants, c‐src, c‐srcNI, and c‐srcNI + Nll. The expression of these transcripts was analysed by PCR amplification of fragments covering the mini‐exons Nl and Nll of the corresponding cDNAs. The PCR products were analysed by Southern hybridization and characterized by determination of their sequences. Neuroblastomas, paragangliomas, retinoblastomas and the phaeochromocytomas expressed neuronal c‐src splice variants. However, whereas neuroblastomas and retinoblastomas contained all 3 transcripts, the phaeochromocytomas and paragangliomas expressed, with 2 exceptions, only the c‐src and the c‐srcNI + Nll mRNA species. To assess whether neuroblastomas display adrenal chromaffin characteristics, they were analysed regarding expression of the chromaffin marker enzyme, phenylethanolamine‐N‐methyl transferase. Whereas phaeochromocytomas were positive, all neuroblastomas were immunochemically negative for this enzyme. These results and the c‐src expression profile suggest that neuroblastomas, including those with an adrenal location, do not originate from the adrenal chromaffin differentiation lineage. The data further suggest neuronal c‐srcNI mRNA as a marker for sympathetic neuronal cells of the sympatho‐adrenal lineage. © 1995 Wiley‐Liss, Inc.
Chromosome abnormalities in benign hyperproliferative disorders of epithelial and stromal breast tissue
Authors: Dietrich Claudia U. , Pandis Nikos , Teixeira Manuel R. , Bardi Georgia , Gerdes Anne‐Marie , Andersen Johan A. , Heim Sverre
Published: February 2007

Cytogenetic analysis of short‐term cultures from 15 cases of benign proliferative breast disease (PBD), 10 diffuse PBD and 5 papillomas, and 15 fibroadenomas of the breast revealed clonal chromosome abnormalities in 7 diffuse PBD lesions, 4 papillomas and 5 fibroadenomas. The remaining 14 cases had a normal female chromosome complement. Cytogenetically unrelated abnormal clones were seen in 4 fibroadenomas and 2 PBDs. A single abnormal clone was found in 9 PBDs and I fibroadenoma. Three clonal abnormalities were seen as recurrent changes in 6 cases, namely interstitial deletions of 3p with 3p 12–14 as the minimally common deleted segment (in I papilloma, I diffuse PBD with atypia and I mixed‐pattern lesion with both papilloma and atypical diffuse PBD features), r(9)(p24q34) (in I diffuse PBD and I fibroadenoma), and del(l)(q12) (again in I diffuse PBD and I fibroadenoma). Intriguingly, 6 of the 16 abnormal cases had chromosome changes that have been seen repeatedly as primary abnormalities in breast carcinomas: der(16)t(l;16)(q10;p10), del(3)(p12p14), and del(l)(q12). We conclude that some of the chromosome anomalies frequently found in breast carcinomas are also present in PBD and fibroadenomas. These aberrations may be accepted as early, neoplasia‐relevant mutations. However, they do not seem to be sufficient by themselves to unleash a malignant process. © 1995 Wiley‐Liss, Inc.
nmb, a novel gene, is expressed in low‐metastatic human melanoma cell lines and xenografts
Authors: Weterman Marian A. J. , Ajubi Nasser , van Dinter Irma M. R. , Degen Winfried G. J. , van Muijen Goos N. P. , Ruiter Dirk J. , Bloemers Henri P. J.
Published: February 2007

From a subtractive cDNA library, we isolated several cDNA clones which showed differential expression between highly and lowly metastatic human melanoma cell lines. One clone, designated nmb, showed preferential expression in the low‐meta‐static cell lines and was chosen for further characterization. Sequence analysis revealed that this clone represents a novel gene, encoding a putative transmembrane glycoprotein which showed the highest homology to the precursor of pMEL17, a melanocyte‐specific protein. nmb RNA expression was absent in most tumor‐cell lines tested and not restricted to the melanocytic lineage. Transfection of a partial nmb cDNA into a highly metastatic melanoma cell line (BLM) resulted, in 2 of 3 transfectants, in slower subcutaneous tumor growth and, in 1 of 3 transfectants, in reduction of the potential for spontaneous metastasis in nude mice. © 1995 Wiley‐Liss, Inc.