bluuebunny/crossref_metadata_2025 huggingface dataset:
HLA restriction and T‐cell‐receptor Vβ gene expression of cytotoxic T lymphocytes reactive with human squamous‐cell carcinoma of the head and neck
Authors: Yasumura Satoshi , Weidmann Eckhart , Hirabayashi Hideki , Johnson Jonas T. , Herberman Ronald B. , Whiteside Theresa L.
Published: February 2007

A human cytotoxic‐T‐lymphocyte (CTL) line capable of killing autologous tumor (AuTu) cell targets was established from peripheral‐blood lymphocytes of a patient with squamous‐cell carcinoma of the tongue. The cultured CTL were CD3+CD8+CDIIb‐HLA‐DR+T cell receptor (TCR) α/β+. When tested in 4‐hr 51Cr‐release assays against various lines of squamous‐cell carcinoma of the head and neck (SCCHN) and a variety of non‐squamous human tumor and normal cell targets, the CTL were found to lyse the autologous SCCHN cell line (PCI50) and 7 allogeneic SCCHN lines: PCI‐1, ‐2, ‐4A, ‐4B, ‐13, ‐30 and ‐38. Of these tumor cell lines, PCI‐13, ‐30 and ‐38 shared HLA‐A2 locus with the AuTu, PC1‐50, while PCI‐4A and ‐4B shared HLA‐B44 with AuTu. Lysis of AuTu (A2+B44+), PCI‐13 (A2+B44‐) and PCI‐4B (A2‐ B44+) by the CTL was efficiently inhibited by monoclonal antibodies (MAbs) to CD3, CD8, TCRα/β or the major‐histocompatibility‐complex(MHC)‐class‐l antigens. MAbs to HLA‐A2 antigens inhibited lysis of PCI‐50 or PCI‐13 targets by the CTL. In cold‐target inhibition assays, unlabeled PCI‐4B or PCI‐13 cells inhibited CTL lysis of AuTu targets. The CTL incubated in the presence of the HLA‐A2+ SCCHN PCI‐50 or ‐13, but not an HLA‐A2+ gastric carcinoma, produced TNF‐α, IFN‐γ and GM‐CSF. The CTL were tested for their TCR Vβ gene expression by polymerase chain reaction (PCR). At week 10 in culture, the time of the highest AuTu cytotoxicity mediated by the CTL line, Vβ was expressed by 26% of T cells. Three clones, obtained by limiting dilution from 10‐week CTL and selected for high cytotoxicity against AuTu, were found to be Vβ+. Further analysis of the specificity of these clones indicated lytic activity against PCI‐13 (A2+B44‐), but not PCI‐4B (A2‐B44+) targets. In 16‐week cultures, which retained AuTu cytotoxicity as well as Vβ expression, TCR Vβ2 was also expressed at high frequency (29%), and AuTu‐reactive clones were found to be Vβ2+. Our results indicate that at least 2 different CTL populations (Vβ2+ and β2+) are able to recognize SCCHN‐associated antigen(s) and that the Vβ6+ T cells are HLA‐A2 restricted, while Vβ6+ T cells are HLA‐A2 restricted, while Vβ2+ T cells may be HLA‐ B44 restricted. © 1994 Wiley‐Liss, Inc.
A prospective study of weight, body mass index and other anthropometric measurements in relation to site‐specific cancers
Authors: Chyou Po‐Huang , Nomura Abraham M. Y. , Stemmermann Grant N.
Published: February 2007

The association of weight, body mass index and other anthropometric measurements with cancer was investigated in a cohort of 7,840 men, examined and interviewed from 1965‐1968 in Hawaii. After 23 years of follow‐up, histologically confirmed incident cases of prostate (n = 306), colon (n = 289), lung (n = 236), stomach (n = 229) and rectal (n = 108) cancer were identified. Body weight was positively associated with prostate cancer. This direct association was stronger for cases diagnosed II or more years after examination than for those diagnosed earlier. A similar pattern was also present for the risk of colon cancer,in association with weight and body mass index. For lung cancer, increased subscapular and triceps skinfold thickness were each associated with decreased risk with adjustment for cigarette smoking, but the inverse association did not persist as the time interval from exam to cancer diagnosis lengthened. There was no significant association between anthropometric measurements and stomach or rectal cancer. © 1994 Wiley‐Liss, Inc.
Intestinal metaplasia types and the risk of gastric cancer: A cohort study in Slovenia
Authors: Filipe M. Isabel , Muñoz Nubia , Matko Ivan , Kato Ikuko , Pompe‐Kirn Vera , Jutersek Albert , Teuchmann Sibylle , Benz Maja , Prijon T.
Published: February 2007

Between 1967 and 1976, 1,525 Slovenian patients with a histological diagnosis of intestinal metaplasia (IM) were classified according to subtype of IM based on morphology and mucin staining; 518 cases were diagnosed with type I, 197 with type II and 275 with type III, but in 291 the diagnosis of IM was not confirmed. Patients who had developed cancer or died up to 1986 were identified by record linkage at the Slovenia Cancer Registry and the Central Population Registry in Slovenia. A total of 34 incident cases of gastric cancer occurring at least 6 months after the diagnosis of IM were identified. The standardised incidence ratio (SIR) for stomach cancer was 2.23 in the whole cohort. It was highest for IM type III, followed by type II and IM‐unconfirmed, but not increased for type I. The relative risk (RR) of developing gastric cancer based on Cox's proportional hazards model was 2.14 for type II and 4.58 for type III, compared with type I. The RR was especially increased for a subgroup of type III secreting sulphomucins in their goblet cells in comparison with types I‐II negative to sulphomucins. Our results confirm that subtyping of IM is useful for identifying individuals at high risk for gastric cancer. © 1994 Wiley‐Liss, Inc.
Cea expression of colorectal adenocarcinomas is correlated with their resistance against lak‐cell lysis
Authors: Kammerer R. , von Kleist Sabine
Published: February 2007

We have investigated the in vitro interaction of LAK cells with 20 different human tumor cell lines freshly isolated from colorectal carcinoma (CRC) specimens. Three steps of LAK cell/tumor cell interaction, namely adherence, infiltration and lysis, have been studied. All are important for the cytotoxic effect of LAK cells against solid tumors: LAK‐cell adherence was studied on tumor‐cell monolayer cultures, the infiltration capacity of LAK cells using tumor‐cell spheroids and the resulting cytotoxic effects of LAK cells against CRC cells grown as spheroids or monolayers. Finally, we correlated the degree of lysis of the CRC cells with their carcinoembryonic antigen (CEA) production, the secretion of which varied in a broad range from not detectable to 2,800 ng/day in culture medium. Cytotoxicity experiments showed a good correlation between CEA expression of CRC cells and their resistance against allogeneic LAK cells. Spheroids of CEA‐producing cells prevented LAK‐cell infiltration resulting in high resistance against LAK‐cell lysis. Adhesion of LAK cells on monolayers of CEA‐expressing CRC cells was minimal. Our results indicate that CEA expression may be an escape mechanism protecting colon carcinoma cells from an attack by cytotoxic cells. © 1994 Wiley‐Liss, Inc.
Expression of APO‐1 (CD95), a member of the NGF/TNF receptor superfamily, in normal and neoplastic colon epithelium
Authors: Möller Peter , Koretz Karin , Leithäuser Frank , Brüderlein Silke , Henne Christof , Quentmeier Armin , Krammer Peter H.
Published: February 2007

APO‐I is a 48‐kDa cell‐membrane protein identical to the Fas antigen now designated CD95. It is a member of the NGF/TNF receptor superfamily. Anti‐APO‐I monoclonal antibody induces apoptosis in a variety of cell types expressing this antigen. We immunohistochemically investigated APO‐I expression in normal colon mucosa, 20 adenomas, 258 colon carcinomas and 10 liver metastases and carried out in vitro studies using a panel of colon‐carcinoma cell lines. Immunohistochemically, APO‐I was regularly expressed at the basolateral membrane of normal colon epithelia. In a minor fraction of colon adenomas and in 39.1% of colon carcinomas APO‐I expression was diminished and in 48.1% of carcinomas, predominantly of the non‐mucinous type, APO‐I expression was completely abrogated. The normal level of APO‐I in carcinomas was correlated with the mucinous type. Reduced/lost APO‐I expression was more frequent in rectal carcinomas. Complete loss of APO‐I was more frequent in tumors that had already metastasized. APO‐I expression in liver metastases essentially corresponded to that of the primary tumors. Comparative analysis with data from previous studies revealed that the mode of APO‐I expression is correlated with that of HLA‐A,B,C./β2m, HLA‐DR, HLA‐D‐associated invariant chain and of the secretory component. Surface expression of APO‐I was heterogeneous in colon‐carcinoma cell lines; SW480 expressed considerable amounts of APO‐I on all cells, while HT‐29 constitutively did less so and only in a minority of cells. Surface density of APO‐I and the fraction of positive cells in HT‐29 was enhanced by interferon‐gamma (IFN‐γ) and, additively, by tumor necrosis factor‐alpha (TNF‐α), whereas in SW480 APO‐I expression was not modulated by these cytokines. We conclude that neoplastic transformation of colon epithelium often leads to a loss of the physiologic, high level of surface APO‐I by giving rise either to a stable lack of APO‐I or to an IFN‐γ/TNF‐α‐sensitive phenotype of inducible APO‐I expression. © 1994 Wiley‐Liss, Inc.
Derivation of androgen‐independent human LNCaP prostatic cancer cell sublines: Role of bone stromal cells
Authors: Wu Hsi‐Chin , Hsieh Jer‐Tsong , Gleave Martin E. , Brown Nicholas M. , Pathak Sen , Chung Leland W. K.
Published: February 2007

A model of human prostate cancer was established to study cellular interaction between prostate cancer and bone stroma in vivo. In this model, subcutaneous co‐injection of 2 non‐tumorigenic human cell lines‐LNCaP, a prostate cancer cell line, and MS, a bone stromal cell line‐into intact adult male mice resulted in formation of carcinomas that secreted prostate‐specific antigen (PSA), a clinically useful human serum prostate cancer marker. In castrated hosts, upon cellular interaction with bone fibroblasts, we observed the progression of these tumors from an androgen‐dependent (AD) to an androgen‐independent state (AI). We derived 4 LNCaP cell sublines from the chimeric LNCaP/MS tumors: the M subline from intact hosts and the C4, C4‐2 and C5 sublines from castrated hosts. The LNCaP sublines had chromosomal markers similar to those of the parental LNCaP cells and distinctly different from those of the MS bone stromal cell line. Although the parental and derived cell lines expressed similar steady‐state levels of orni‐thine decarboxylase transcript, the sublines expressed 5‐ to 10‐fold higher basal steady‐state levels of PSA transcript than did the parental LNCaP cell line. The LNCaP sublines formed 13‐ to 26‐fold more soft‐agar colonies than the parental LNCaP cell line. The sublines became tumorigenic, yielding an incidence of tumors in intact athymic mice of 7‐75%. The LNCaP sublines C4 and C5 (but not the parental and M cell line) formed tumors in castrated hosts when co‐injected with bone fibroblasts. A second‐generation LNCaP subline, C4‐2, was derived from a chimeric tumor induced by co‐inoculating castrated mouse with C4 cells and MS cells. We found that C4‐2 subline was tumorigenic when inoculated into castrated hosts in the absence of inductive fibroblasts. Moreover, C4‐2 was the only subline capable of forming soft‐agar colonies when cultured in serum‐free medium. In comparison with the parental LNCaP cells, the C4‐2 subline expressed lower steady‐state levels of androgen receptor (AR) protein and mRNA transcript and lost its androgen responsiveness in vitro. Our results suggest that certain genetic traits of prostate cancer cells may be selected or altered through an “adaptive” mechanism that involves cellular interaction with the bone stromal cells. © 1994 Wiley‐Liss, Inc.
Isolation of naturally processed peptides recognized by cytolytic T lymphocytes (CTL) on human melanoma cells in association with HLA‐A2.1
Authors: Wölfel Thomas , Schneider Jörg , Zum Büschenfelde Karl‐Hermann Meyer , Rammensee Hans‐Georg , Rötzschke Olaf , Falk Kirsten
Published: February 2007

Cytolytic T lymphocyte (CTL) clones have previously been derived from peripheral blood of melanoma patient SK29(AV). They lyse autologous melanoma cells but not autologous Epstein‐Barr virus (EBV)‐transformed B lymphocytes. Immunoselection experiments indicate that these CTL clones recognize 4 different antigens (Aa, Ab, B, C) in association with a single HLA restriction element, HLA‐A2.1. While the expression of antigens B and C appears to be confined to SK29‐melanoma cells, antigens Aa and Ab are shared by a high proportion of allogeneic HLA‐A2‐positive melanoma lines. HLA‐A2.1 and total HLA class 1 molecules have now been purified from SK29‐melanoma cells using affinity chromatography and associated peptides have been eluted. Peptide pools eluted from HLA‐A2.1 and total class 1 were separated by reversed phase high performance liquid chromatography (HPLC). Individual HPLC fractions were tested for their ability to sensitize target cells for recognition by SK29‐CTL clones. The presence of antigens Aa, Ab, B and C was detected in distinct HPLC fractions that were identical for both peptide pools. As target for detection of peptide antigens in HPLC fractions, the use of the HLA‐A2.1‐positive antigen processing mutant cell line CEM x 721.174.T2 (T2), pre‐incubated with anti‐HLA‐A2 monoclonal antibody (MAb) MA2.1, was shown to be essential. Single‐peak target‐sensitizing activity was found for antigens Ab and B, whereas multi‐peak sensitizing activity was reproducibly detected for antigens Aa and C. We reason that at least some of these melanoma peptide antigens might occur in biochemically distinct isoforms. © 1994 Wiley‐Liss, Inc.
Association between HLA‐DQB1 alleles and risk for cervical cancer in African‐American women
Authors: Gregoire Lucie , Lawrence W. Dwayne , Kukuruga Debra , Eisenbrey A. Bradley , Lancaster Wayne D.
Published: February 2007

Squamous‐cell carcinoma of the cervix and its precursor lesions are associated with human papillomavirus (HPV) infection. Epidemiological studies indicate that HPV infection in itself is not sufficient for cervical‐cancer induction, suggesting that other factors contribute to carcinogenesis. We have investigated the potential role of host genetic background as one such factor. We screened a series of squamous‐cell carcinomas of the cervix for HLA‐class‐II DQBI* alleles by the polymerase chain reaction and site‐specific oligonucleotide probe hybridization and for HPV type from African‐American women using a local, ethnically matched control panel. Statistically significant associations for increase in relative risk for cervical cancer were seen for DQBI*0303 and DQBI*0604. DQBI*0201 and the hetero‐zygote DQBI*0301/*0501 showed a decrease in relative risk for cervical cancer. HPV typing revealed no association between virus type and DQBI alleles. Our results confirm other studies showing an increase in relative risk for cervical cancer associated with HLA‐DQ3 alleles in Caucasians. © 1994 Wiley‐Liss, Inc.
Epidermal‐growth‐factor‐receptor expression is associated with rapid tumor proliferation in bladder cancer
Authors: Sauter G. , Haley J. , Chew K. , Kerschmann R. , Moore D. , Carroll P. , Moch H. , Gudat F. , Mihatsch M. J. , Waldman F.
Published: February 2007

Epidermal‐growth‐factor‐receptor (EGF‐r) expression has been proposed as a prognostic marker in bladder cancer and is associated with rapid proliferation in cell lines. Ninety‐three fresh and 74 formal in‐fixed bladder tumors were examined by fluorescence in situ hybridization (FISH) and immunohistochemistry to assess the relationship between EGF‐r expression and proliferation as well at the prevalence of epidermal‐growth‐factor‐receptor (EGF‐r) gene amplification. EGF‐r expression was strongly associated with BUdr labeling index, grade and stage. EGF‐r expression emerged as a stronger predictor of tumor proliferation than grade or stage in analysis of variance. Rapid tumor proliferation might be responsible for bad prognosis reported in EGF‐r positive bladder tumors. Also chromosome 7 copy number was associated with grade and stage. EGF‐r gene amplification was uncommon (5 of 107 tumors). However, FISH analysis allowed characterization of the pattern of amplification, with clustering of signals suggestive of intrachromosomal amplification more common than diffuse distribution consistent with extrachromosomal amplification. © 1994 Wiley‐Liss, Inc.
Molecular mechanisms possibly affecting WT1 function in human ovarian tumors
Authors: Viel Alessandra , Giannini Franca , Capozzi Eugenia , Canzonieri Vincenzo , Scarabelli Carlo , Gloghini Annunziata , Boiocchi Mauro
Published: February 2007

The frequent allelic deletions observed on the short arm of chromosome 11 in ovarian tumors suggest that the WTI gene, a proposed tumor‐suppressor gene located on chromosome 11 p 13 and expressed in the human fetal genitourinary system, may contribute to the development of ovarian neoplasms. Structural and sequence analysis of the entire coding portions of the WTI gene did not reveal any abnormalities in the 20 ovarian tumor specimens (13 of which showed 11p13 allelic deletions) and 5 cell lines which we analyzed. These findings invalidate the hypothesis that the WTI gene functions as a classical tumor‐suppressor gene in ovarian tumorigenesis and suggest that a different recessive oncogene may be “exposed” by the observed 11 p 13 allelic deletions. Expression analysis showed that the WTI gene was transcriptionally active in all the tumors tested, but considerable variations in the mRNA levels were found. This apparent variability, which should be confirmed at the cellular level in the tumor specimens, was also observed in the ovarian tumor‐cell lines. Finally, WTI expression data were evaluated in conjunction with immunohistochemical data on p53. The possible functional effects of altered WTI mRNA expression in ovarian tumors are discussed, taking into account the potential WTI/p53 protein interaction. © 1994 Wiley‐Liss, Inc.
Cellular and karyotypic characterization of two doxorubicin resistant cell lines isolated from the same parental human leukemia cell line
Authors: Zalcberg John R. , Hu Xiu F. , Wall Dominic M. , Mirski Shelagh , Cole Susan , Nadalin Gabrielle , de Luise Mario , Parkin John D. , Vrazas Vickie , Campbell Lynda , Kantharidis Phillip
Published: February 2007

Separate mechanisms underlying the multidrug resistant (MDR) phenotype were identified in 2 independent approaches to select tumour cells resistant to low concentrations of doxorubicin (Dox) from the sensitive T cell leukemia cell line CCRF‐CEM. The CEM/A7 cell line was selected at an initial concentration of 0.005 μg/ml of Dox and maintained at 0.07 μg/ml. In contrast, the CEM/A5 line was selected using an initial concentration of 0.01 μg/ml and maintained in Dox at a concentration of 0.05 μg/ml. P‐glycoprotein expression was demonstrated in the CEM/A7 line but not the CEM/A5 line. Amplification of the mdr1 gene was not observed in the CEM/A7 cell line. Both cell lines showed cross‐resistance to a number of structurally unrelated cytotoxic drugs including anthracyclines and etoposide (VP‐16), although only the CEM/A7 line was cross resistant to Vinca alkaloids. Immunoblots of total cell lysates of the CEM/A5 line have revealed almost undetectable levels of topoisomerase 11 α and β in this line. Cytogenetic analyses of both lines revealed numerous karyotypk abnormalities which were present in the parental cell line as well as both resistant cell lines. The CEM/A7 line also demonstrated a duplication of part of the long arm of chromosome 7 which included the region containing the mdr1 gene, a finding not seen in the parental or CEM/AS line. CEM/AS, however, demonstrated an abnormality of chromosome 7, outside the region of the mdr1 gene, and it also contained a deletion of the short arm of chromosome 2. Abnormalities in this latter region of genome have been associated with non‐P‐glycoprotein‐mediated MDR. © 1994 Wiley‐Liss, Inc.
Inhibition of tumour promotion by various palm‐oil tocotrienols
Authors: Goh S. H. , Hew N. F. , Norhanom A. W. , Yadav M.
Published: February 2007

Inhibition of tumour promotion by various vitamin E compounds (tocopherols and tocotrienols) and some of their dimers was examined by an in vitro assay utilizing the activation of Epstein‐Barr virus (EBV) early antigen (EA) expression in EBV‐genome‐carrying human lymphoblastoid cells. The results reveal that γ‐ and δ‐tocotrienols derived from palm oil exhibit a strong activity against tumour promotion by inhibiting EBV EA expression in Raji cells induced by 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA). However, α‐ and γ‐tocopherols and dimers of γ‐tocotrienol or γ‐tocopherol lack this activity. © 1994 Wiley‐Liss, Inc.
The pattern of cytokine gene expression in freshly excised human metastatic melanoma suggests a state of reversible anergy of tumor‐infiltrating lymphocytes
Authors: Lüscher Urs , Filgueira Luis , Juretic Antonio , Zuber Markus , Lüscher Nicolas J. , Heberer Michael , Spagnoli Giulio C.
Published: February 2007

Expression of an extended panel of cytokine genes was investigated by reverse polymerase chain reaction (PCR) in 10 freshly excised melanoma metastases infiltrated by lymphocytes (TIL). cDNA encoding for CD3‐δ and tyrosinase could be amplified in all samples, confirming the presence of T lymphocytes and melanoma cells. Cytokine genes possibly transcribed by both cell types, such as GM‐CSF, IL‐6 and IL‐10 could be amplified from 5, 2 and 2 samples respectively. In contrast, IL‐β and TNF‐α mRNA were never detectable, IL‐1α, IL‐3 and IL‐7 mRNA could be observed only in one case each. Transcripts encoding for TGF‐β1 were observed in 8 samples, while TGF‐β2 and 3 mRNA were detectable in only 2 specimens. mRNA encoding for cytokine genes typically transcribed by antigen‐stimulated T lymphocytes, such as IL‐2, IL‐4 and IFN‐7 were rarely or never detectable (none, none and 1 of the samples respectively). In one case, where no cytokine gene transcription was detectable at the time of surgery, we addressed the question of the antigenicity of the tumor and of the functional competence of TIL. A primary tumor cell line was generated and cultured TIL were induced to transcribe IL‐2 and IFN‐γ genes by incubation with the autologous irradiated tumor cell line, but not with autologous EBV‐transformed cells. In these conditions, tumor‐specific cytotoxic T lymphocytes (CTL) could be generated only after 3 weekly re‐stimulations. In contrast, if autologous irradiated EBV‐transformed cells were added to the cultures, specific CTL could be detected after one single tumor stimulation. Thus, signs of active responsiveness in terms of lymphokine gene mRNA are seldom detectable in melanoma metastases. Tumor‐specific responses, however, including IL‐2 and IFN‐γ gene expression and generation of CTL can be produced in vitro from specimens in which no cytokine gene mRNA is detectable ex vivo.
Biological‐clinical significance of selective loss of HLA‐class‐I allelic product expression in squamous‐cell carcinoma of the uterine cervix
Authors: Honma Shigeru , Tsukada Seiji , Honda Shuji , Nakamura Minoru , Takakuwa Koichi , Maruhashi Toshihiro , Kodama Shojj , Kanazawa Koji , Takahashi Takeshi , Tanaka Kenichi
Published: February 2007

To determine possible correlations between the selective loss of HLA‐class‐l allelic forms on neoplastic cells and their biological‐clinical characteristics, 89 squamous‐cell carcinomas of the uterine cervix were evaluated immunohistochemically using monomorphic and polymorphic antibodies against HLA‐A, ‐B, and ‐C molecules and analyzed clinico‐pathologically. Four of the carcinomas exhibited a lack of detectable class‐l heavy‐chain expression associated with β2‐microglobulin. In 19 of 42 HLA‐A2‐positive patients, tumor cells revealed loss of the HLA‐A2 allelic product. Loss of HLA‐B7 and/or 40 (B7/40) allelic product(s) on tumor cells was observed in 12 of 2S HLA‐B7/40‐positive cases. These alterations did not correlate with patient age, clinical stage (FIGO) of the disease, histological sub‐type (WHO) or depth of cervical invasion. However, a statistically significant correlation was observed between lymph‐node metastases and selective loss of HLA‐B7/40 allelic produces), but not with HLA‐A2 allelic product on cancer cells of the primary lesion. Our results indicate that selective loss of certain HLA‐class‐l alleles on neoplastic cells can influence the nodal metastatic potential and suggest that these 2 class‐l molecules may have different immune functions as restriction elements in T‐cell‐mediated cytotoxicity.
Tumor‐derived transforming growth factor‐β1 and interleukin‐6 are chemotactic for lymphokine‐activated killer cells
Authors: Delens Nadia , Torreele Els , Savelkool Huub , de Baetselier Patrick , Bouwens Luc
Published: February 2007

Adherent lymphokine‐activated killer (A‐LAK) cells are purified IL‐2 activated natural killer (NK) cells with potent anti‐tumor cytotoxic activity. They have been used in the adoptive immunotherapy of metastatic canters. However, it has been shown that intravenously transferred LAK cells have a poor horning capacity to tumor sites. For the present study, the effects of tumor‐derived factors on the in vitro migratory capacity of A‐LAK cells was investigated. In a micropore migration assay the conditioned medium from 3LL Lewis lung carcinoma cell cultures was found to exert a strong chemotactic, but not chemokinetic effect on A‐LAK cells. This effect was partially inhibited by neutralizing antibodies against the cytokines TGF‐β1 and IL‐6. A combination of the 2 antibodies completely suppressed the chemotactic activity of tumor‐cell‐conditioned medium. Purified TGF‐β1 and recombinant IL‐6 were chemotactic for A‐LAK cells. Biological activities of both cytokines were detectable in the tumor‐cell‐conditioned medium. The in vivo relevance of these findings, with respect to tissue infiltration of NK cells and LAK cells in inflammation or cancer, remains to be elucidated.
Mitogenic activity of procathepsin D purified from conditioned medium of breast‐cancer cells by affinity chromatography on pepstatinyl agarose
Authors: Stewart A. J. , Piggott N. H. , May F. E. B. , Westley B. R.
Published: February 2007

A simple procedure for the affinity purification of procathepsin D from tissue culture medium conditioned by breast‐cancer cells is described. This procedure yielded 2 μg of procathepsin D/100 ml medium. The procathepsin D was approximately 95% pure as judged by silver staining of polyactylamide gels, the major contaminant being mature cathepsin D. The ability of procathepsin D to stimulate the proliferation of oestrogen‐responsive MCF‐7 breast cancer cells was determined. The purified procathepsin D had no mitogenic effect alone or in combination with oestradiol or other growth factors. These data suggest that procathepsin D does not act as an oestrogen‐regulated autocrine growth factor for malignant breast epithelial calls.
Regulation of fibronectin and laminin receptor expression, fibronectin and laminin secretion in human colon cancer cells by transforming growth factor‐β1
Authors: Huang Shuang , Chakrabarty Subhas
Published: February 2007

Transforming growth factor (TGF)‐β1, modulates the expression of extracellular matrix (ECM) glycoproteins, fibronectin and laminin and the adhesion of Moser colon cancer cells to these glycoproteins. Since adhesion can be altered through expression of cell‐surface receptors, binding affinities of adhesion molecules for receptors, or both, we investigated the effect of TGF‐β1, on the binding properties of fibronectin and laminin to their cell‐surface receptors by saturation binding and Scatchard analyses using radiolabeled fibronectin and laminin. Fibronectin bound to its cell‐surface receptor with high affinity (Kd = 1.25 × 10−9 M), Moser cells had approximately 7.1 × 104 fibronectin‐binding sites per cell. TGF‐β1 treatment rapidly up‐modulated the number of cell‐surface fibronectin‐binding sites by 1.9‐fold. The binding affinity of fibronectin for the receptor, however, was not altered. Laminin was found to bind to a higher‐affinity and a lower‐affinity receptor. Moser cells expressed approximately 1.1 × 103 higher‐affinity laminin‐binding sites and approximately 3.1 × 10 lower‐affinity‐binding sites per cell. TGF‐β1 rapidly increased the expression of the higher‐affinity sites 3‐fold and the lower‐affinity sites 5‐fold. The binding affinity of both the higher‐affinity and lower‐affinity laminin receptors increased 3‐fold after 2 and 6 hr of TGF‐β1 treatment respectively. Concurrent with receptor modulation, TGF‐β1 induced the secretion of fibronectin and laminin from Moser cells. Northern hybridization analyses showed a concurrent stimulation of the expression of the mRNAs for ligands (fibronectin and laminin) and the mRNAs for the integrin species of the fibronectin and laminin receptors (α5 and α6 subunits). Thus the production of fibronectin and laminin and the expression of their receptors were tightly co‐regulated by TGF‐β1.
Effect of B‐lymphocyte‐ and NPC‐derived EBV‐1 gene expression on in vitro growth and differentiation of human epithelial cells
Authors: Zheng Xi , Yuan Fang , Hu Lifu , Chen Fu , Klein George , Christensson Birger
Published: February 2007

The effect of expression of the Epstein‐Barr‐virus (EBV) latent membrane protein (LMP1) derived from B‐lymphocytes (B) and nasopharyngeal carcinoma (NPC) (C) on the in vitro growth and differentiation of a human keratinocyte line, Rhek‐1, was analyzed in clonal growth and in in vitro differentiation assays. In contrast to the polygonal parental cells, the B‐LMP1‐expressing sublines were spindle‐shaped while the C‐LMP1‐expressing cells were pleomorphic. Both B‐ and C‐LMP1‐expressing sublines showed increased proliferation as evidenced by: (1) higher colony‐forming efficiency (CFE) and larger colony size at reduced serum levels; (2) an increased number of epithelial cell layers formed in the air‐liquid‐interface culture system and (3) increased expression of proliferative cell nuclear antigen (PCNA). At low serum concentration, the C‐LMP1‐expressing sublines formed larger colonies than those expressing B‐LMP1. In the air‐liquid‐interface culture system, both B‐ and C‐LMP1‐expressing lines showed reduced epithelial differentiation resulting in reduced stratification and reduced involucrin expression similar to those of the cancer cell line, Siha. The results of the present study indicate that the expression of LMP1 in human keratinocytes is associated with morphological transformation and predisposes these cells to a more neoplastic phenotype. The structural difference between the 2 genes responsible for the functional differences and transforming ability will be pinpointed in further experiments.
Mothers' active and passive smoking during pregnancy and risk of brain tumours in children
Authors: Filippini G. , Farinotfi M. , Lovicu G. , Maisonneuve P. , Boyle P.
Published: February 2007

As part of a collaborative study of risk factors for childhood brain tumours, the effects of the mother's smoking and her potential for passive smoking exposure during the pregnancy were assessed in a case‐control study. Parents of 91 cases and 321 population controls from Northern Italy, matched for age, sex and residence, were interviewed about their lifetime smoking habits. Mother's smoking during pregnancy was associated with an odds ratio (OR) of 1.7 (95% CI 0.8, 3.8) of brain tumour in her child although this was not statistically significant. Among non‐smoking mothers, the risk for light and heavy exposure to passive smoking was 1.7(0.8, 3.6) and 2.2 (1.1, 4.5) respectively, and a statistically significant dose‐response relationship was found (p trend = 0.02). These results must be interpreted within the constraints of the relatively small sample size and the likely misclassification produced by the difference between the potential for exposure to passive smoke and the true exposure. However, they add another piece of information to the growing body of evidence available about the health consequences both of active and of passive smoking and highlight the need for more information about this putative association. © 1994 Wiley‐Liss, Inc.
Gastric cancer in Iceland: A retrospective study of resected gastric specimens in a high‐risk country during 30 years (1960–1989)
Authors: Jónasson Lárus , Hallgrímsson Jónas , Sigvaldason Helgi , Ólafsdóttir Gudrídur , Tulinius Hrafn
Published: February 2007

The world‐wide incidence of gastric cancer is decreasing, especially in high‐risk populations such as the Icelanders. We examined retrospectively 1,040 specimens of resected gastric cancers from a 30‐year period, 1960–1989. The decrease in incidence in both sexes involved mainly the largest histological group, the intestinal‐type tumours. In males there was also a decrease in diffuse tumours but in females these remained relatively unchanged. Tumours of the antrum and the corpus decreased in both sexes. In males there was a significant increase in tumours of the cardia and most of these were of the intestinal type. In females there was a minor increase in tumours of the cardia, all of which were of the intestinal type. An increase in incidence of tumours of the cardia concomitant with a decrease in incidence of tumours in other parts of the stomach suggests a difference in aetiological factors. The death risk for patients with tumours of the cardia was 59% higher than that for those with tumours in other parts. The death risk following gastric resection was not significantly different when patients with intestinal‐type tumours and diffuse tumours were compared. © 1994 Wiley‐Liss, Inc.
Metastatic renal‐cell carcinoma patients treated with interleukin 2 or interleukin 2 plus interferon γ: Immunological monitoring
Authors: Farace Françoisc , Pallardy Marc , Angevin Eric , Hercend Thierry , Escudier Bernard , Triebel Frédéric
Published: February 2007

We investigated the biological response of 73 patients with metastatic renal‐cell carcinoma (MRCC) treated by repetitive weekly cycles of high‐dose interleukin 2 (IL‐2) (protocol 1, 40 patients) or IL‐2 plus interferon‐gamma (IFN‐γ) (protocol 2, 33 patients). The objectives of this study were (i) to evaluate the effects of this IL‐2 administration schedule on biological response, (ii) to compare the effects of IL‐2 alone with those of IL‐2 plus IFN‐γ, (iii) to search for any correlation between certain biological marker values and the clinical response to treatment. Mean CD56+ lymphocyte counts (i. e., NK cells) were significantly higher than those of CD3+ cells in the 2 protocols and a subpopulation of CD56bright cells in protocol I was found to be preferentially expanded in vivo. Cytotoxic activity against K562 and Daudi cell lines as well as TNF‐α and sTNF‐αR (but not IL‐6) significantly increased following treatment. Comparison of the data obtained from patients treated with IL‐2 alone vs. IL‐2 plus IFN‐γ did not show any significant changes except for eosinophilia (higher in protocol I). Therefore, addition of IFN‐γ did not affect either lymphocyte distribution or non‐MHC‐restricted cytotoxicity in vivo. No difference in cell subpopulation or cytotoxicity was detected between responders and non‐responders. Pre‐treatment sTNF‐αR concentration, in contrast to IL‐6 and TNF‐α, was significantly higher in progressive than in stable and responder groups, suggesting that this parameter may be predictive of the clinical response. © 1994 Wiley‐Liss, Inc.
Immunohistochemical evaluation of P‐glycoprotein in human malignancies by monoclonal antibody MC57
Authors: Leonardo Eugenio , Valente Guido , Cappia Susanna , Cianfriglia Maurizio , Arancia Giuseppe , Meschini Stefania , Scagliotti Giorgio V.
Published: February 2007

P‐glycoprotein expression was analyzed on 137 formalinfixed, paraffin‐embedded human tumours by monoclonal antibody (MAb) MC57. This MAb recognizes an extracellular human specific P‐glycoprotein epitope and defines their multidrug resistance (MDR) phenotype by its binding on cells. Immunohistochemistry indicated that this MAb reacted in human cells and tissues in the same pattern as that found with other MAbs to P‐glycoprotein. However, the present extensive study demonstrated that MAb MC57 is a highly specific reagent for the evaluation of an extracellular P‐glycoprotein epitope preserved after fixation procedures and that this MAb is available to assess P‐glycoprotein expression in routinely processed human tumour specimens. © 1994 Wiley‐Liss, Inc.
Endogenous TGF‐β contributes to the induction of the EBV lytic cycle in two burkitt lymphoma cell lines
Authors: di Renzo Livia , Alttok Aysin , Klein George , Klein Eva
Published: February 2007

A low proportion of cells in the BL lines P3HR‐1 and Akata enter spontaneously into the EBV lytic cycle, detectable by the expression of early antigens (EA). We found that both lines produce the active and inactive forms of TGFß. It was shown earlier that a larger number of cells can be induced to enter the lytic cycle by exposing P3HR‐1 to phorbol esters and n‐butyrate and the surface IgG‐positive Akata cells to anti‐IgG. We now show that the same treatments raise the level of active TGFß release. Exposure to anti‐TGFß antibodies reduced EA induction by 75‐85%. Our results indicate that induction of the viral productive cycle by the above‐mentioned reagents is at least partly dependent on the activation of endogenous TGFß production. © 1994 Wiley‐Liss, Inc.
A staging system for soft‐tissue sarcoma and its evaluation in relation to treatment
Authors: Tomita Y. , Kuratsu S. , Naka N. , Uchida A. , Ono K. , Ohsawa M. , Aozasa K.
Published: February 2007

In order to define the significant factors for a staging system of soft‐tissue sarcomas(STS), histologic and clinical findings in 190 adult patients with localized STS in the extremities and trunk were reviewed. The male‐to‐female ratio was 1.21. The histologic grading of tumors was defined according to the criteria recently proposed by us: tumors were low‐grade in 65 cases, intermediate‐grade in 57 cases and high‐grade in 68 cases. The initial surgical procedure was as follows: intracapsular excision in 9 cases, marginal excision in 104 and wide local excision in 77, including 15 amputations. The mode of treatment was surgery alone (101 patients), surgery and chemotherapy (58), surgery and radiotherapy (22) and surgery and combined chemo‐ and radiotherapy (9). Univariate analysis revealed histologic grade, sex, tumor size and tumor depth to be significant prognostic factors. Multivariate analysis revealed histologic grade to be the only independent factor for prognosis. Significant clinical factors in each histologic grade were then evaluated. In the low‐grade group, local recurrence significantly affected prognosis. Most of the patients with local recurrence had had marginal resection as the initial surgical procedure. No clinical factors affecting prognosis in the intermediate‐grade group could be determined. In the high‐grade group, patients with wide local excision and adjuvant chemotherapy had a better prognosis than those with marginal excision with or without adjuvant chemotherapy and wide local excision without chemotherapy (p=0.09). In conclusion, histologic grade was the only significant factor for the staging of STS. On the basis of our staging system, different modalities of treatment for each grade of STS might be indicated; adequate surgery is essential for the prevention of local recurrence, which resulted in reduced mortality in patients with low‐grade STS. For high‐grade STS, the prevention of distant metastasis by combined extensive surgery and adjuvant chemotherapy may make long‐term survival possible.
International comparison on ras gene mutations in latent prostate carcinoma
Authors: Watanabe Masatoshi , Shiraishi Taizo , Yatani Ryuichi , Nomura Abraham M. Y. , Stemmermann Grant N.
Published: February 2007

Latent carcinomas of the prostate, discovered at autopsy in men with no prior treatment for prostatic disease, were studied for ras proto‐oncogene mutations. Subjects included 21 Japanese, 15 U.S. whites, 15 U.S. blacks, 20 Hawaiian Japanese and 10 Colombians. PCR and sequence‐specific oligonucleotide hybridization identified mutations in 5 Japanese, in 1 Hawaiian Japanese, in 1 U.S. black, in 1 U.S. white and in 3 Colombians. The 5 Japanese tumor samples contained 3 point mutations in codon 12 of K‐ras and 2 mutations in codon 12 of N‐ras respectively. One tumor in a Hawaiian Japanese man also showed a K‐ras point mutation at codon 12. Two Colombians and one U.S. black man had tumors with mutations at codon 61 of H‐ras, while 1 Colombian showed an N‐ras mutation at this codon. The overall frequency of ras gene mutations was low, but point mutations in codon 12 were most common in latent tumors of Japanese, who experienced the lowest incidence and mortality from this tumor. Latent tumors in men from ethnic groups with high mortality and incidence rates showed fewer ras mutations than the Japanese, and these were more likely to involve codon 61. This finding is consistent with prior studies of more aggressive clinical cancers in Japanese men that indicated a higher frequency of mutations at codon 61.
Comparative analysis of mutations in the p53 and K‐ras genes in pancreatic cancer
Authors: Berrozpe Georgina , Schaeffer James , Peinado Miguel Angel , Real Francisco X. , Perucho Manuel
Published: February 2007

Mutations in codon 12 of K‐ras occur in a high proportion of pancreatic cancer cases. Although there is evidence that p53 mutations also occur in this tumor, few studies have been reported to date and no comparison has been made of K‐ras and p53 mutations in the same tissues. Single‐strand conformation polymorphism and sequencing of the PCR products were used to determine mutations in p53 gene; to detect mutations in K‐ras genes, the artificial restriction fragment length polymorphism (RFLP) approach was used. Eight out of 30 tissues from primary pancreas cancer and 3 of 4 samples from metastases showed p53 mutations. Fifteen out of 17 pancreatic cancer cell lines had p53 mutations. In 2 cases, the same p53 mutation was identified in the original tumor and in a tumor‐derived cell line. The majority of p53 mutations were present in exons 5‐9 of the gene. Mutations at codon 12 of the K‐ras gene were identified in 23/32 pancreas cancer tissues and in 14/17 cell lines. There was no relationship between the types of mutation observed in the 2 genes. In conclusion, mutations in K‐ras and p53 genes are common in pancreatic cancer. p53 mutations may occur more frequently in metastatic lesions than in primary tumors, although further work is necessary to investigate this point.
Multiple occurrence of carcinoma in the upper aerodigestive tract associated with esophageal cancer: Reference to smoking, drinking and family history
Authors: Morita Masaru , Kuwano Hiroyuki , Ohno Shinji , Sugimachi Keizo , Seo Yosuke , Tomoda Hirotsugu , Furusawa Motonosuke , Nakashima Tadashi
Published: February 2007

Smoking and drinking habits as well as family history were examined in 143 men with esophageal cancer, including 30 who had associated second cancers of the upper aerodigestive tract (UADT) and 113 who did not. The risk of second cancers of UADT associated with the main lesions of the esophagus was evaluated, using odds ratios (ORs). As a result, the ORs of second cancers for current smoking and drinking were 5.3 and 7.6 respectively. The ORs significantly increased to 12.7 and 14.7 in heavy smokers and in heavy drinkers respectively. Furthermore, the risk of second cancer also significantly increased (8‐fold) in patients who had close relatives with UADT cancer, compared to those without family history of any cancer. However, there were no differences in smoking or drinking habits regardless of family history. Our data thus suggest that a family history of UADT cancer as well as heavy smoking and drinking are clearly associated with multiple occurrence of UADT cancer. Therefore, careful and frequent examination for appearance of any second lesions are required for patients in these high‐risk groups.
Metastasis of mouse T lymphoma cells is controlled by the level of major histocompatibility complex class I H‐2DK antigens
Authors: Driessche Thierry Vanden , Geldhof Anja , Bakkus Marleen , Toussaint‐Demylle Dominique , Brijs Lea , Thielemans Kris , Verschueren Hendrik , De Baetselier Patrick
Published: February 2007

In vivo inoculation of a low metastatic BW 5147 derived T‐cell lymphoma variant into syngeneic mice, had led to the generation of a highly metastatic variant. The shift towards a more metastatic phenotype is accompanied by an increase in major histocompatibility class I H‐2Dk antigen expression. This suggests that H‐2Dk antigens may control the metastatic potential of BW T lymphoma cells. Our present findings indicate that H‐2Dk expression is directly correlated with the metastatic potential of BW cells. We have confirmed such correlation by specifically altering the level of H‐2Dk expression by: I) FACS analysis, 2) IFN‐γ treatment, 3) H‐2Dk gene transfection. Cells sorted for low H‐2Dk expression had a significantly reduced metastatic potential. Induction of H‐2Dk expression on these cells by either IFN‐γ treatment or H‐2Dk gene transfection concomitantly led to increased metastasis. We also assessed metastatic potential of BW cells in irradiated, immunocompromised recipients. Our results show that the immune system is implicated and we further tested which immune effectors are involved, In vivo depletion of natural killer (NK) and CD8+ T‐cells revealed that the difference in metastatic potential of the H‐2Dk variants relies upon an NK‐dependent mechanism, whereas CD8+ T‐cells are not implicated. Our observations suggest that highly metastatic cells, expressing a high level of H‐2Dk antigens are more resistant to NK‐cell‐mediated cytotoxicity in vivo. We have confirmed our in vivo results by in vitro cytotoxicity assays using poly I:C induced NK and IL‐2 activated LAK cells. We conclude that a NK‐dependent mechanism accounts for the association between differential H‐2Dk antigen expression and metastasis.
Analysis of gene amplification and overexpression in human esophageal‐carcinoma cell lines
Authors: Kanda Yuji , Nishiyama Yasuyuki , Shimada Yutaka , Imamura Masayuki , Nomura Hiroshi , Hiai Hiroshi , Fukumoto Manabu
Published: February 2007

Gene amplification/overexpression was analyzed in 23 cell lines derived from human esophageal squamous‐cell‐carcinoma tissues by Southern and Northern hybridizations to c‐myc, c‐erbB, hst‐1 and cyclin‐DI probes. Amplification of the c‐myc gene was observed in 5 cell lines derived from well‐differentiated carcinomas and all of them were accompanied by co‐amplification of other examined oncogenes. The c‐erbB gene was amplified in 3 cell lines. Co‐amplification of hst‐1 and cyclin DI, both of which are located in chromosome 11q13, was found in 9 cell lines. Without exception their amplification was simultaneous and the magnitudes were similar. Their amplification, but not their overexpression, was significantly correlated with poor prognosis in patients from whom the cell lines were established. While hst‐1‐gene expression was not detected, at least 1 of the genes analyzed was overexpressed in 20 cell lines vs. its expression in normal esophageal mucosal tissues. However, gene amplification was not necessarily accompanied by overexpression of the corresponding genes. Expression of the cyclin DI gene, which has been assumed to be a target gene for 11q13 amplification, was not detected in one particular cell line with amplification of 11q13.

These results suggest that the amplification/overexpression of more than I oncogene is involved in the carcinogenic process of esophageal carcinoma and that c‐myc‐gene amplification is associated with a well‐differentiated subtype. There remains a possibility that key oncogenes other than cyclin DI are involved in 11q13 amplification.