bluuebunny/crossref_metadata_2025 huggingface dataset:
Cancer incidence among parents of patients with colorectal cancer
Authors: Søndergaard Jens O. , Bülow Steffen , Lynge Elsebeth
Published: February 2007

To investigate the genetic factor in the development of colorectal cancer, a cohort study was undertaken of parents of patients with this disease. All 1,524 patients, who were diagnosed with colorectal cancer in Denmark in 1982‐84 and were below the age of 60 years, were selected from the National Cancer Register. The parents of these patients were identified from the public population registers. The index persons had a total of 1,478 eligible mothers, of whom 96% were successfully traced, and a total of 1,414 eligible fathers, of whom 96% were traced. These parents were sought in the Cancer Register for cancer cases diagnosed in the period January 1, 1943 to December 31,1986. The incidence rates for the Danish population were used to estimate the expected numbers of colorectal cancer cases among the parents. Both the mothers and the fathers exhibited an increased risk of colorectal cancer. The standardized incidence ratios were 1.62 (95% CI 1.31‐2.01) and 1.87 (95% CI 1.54‐2.27), respectively. In a previous study we found that spouses of patients with colorectal cancer in Denmark did not have an increased risk of this disease. The increased risk found in the present study among the parents of patients therefore indicates that a possible genetic factor is present in the aetiology of colorectal cancer, and that it is of importance in the general population.
Analysis of marek's disease tumor‐associated surface antigen on MDCC‐MSB1‐CLO.18 cells
Authors: Home Masato , Ohashi Kazuhiko , Kodama Hiroshi , Mikami Takeshi
Published: February 2007

The physico‐chemical nature of Marek's disease tumor‐associated surface antigen (MATSA) on Marek's disease (MD) lymphoblastoid tumor cell line (MDCC‐MSBI‐clo.18) was examined by the cellular enzyme‐linked immunosorbent assay (CELISA) and the sandwich enzyme‐linked immunosorbent assay (ELISA) using an anti‐MATSA immune serum or a monoclonal antibody (MAb) 2B9 developed against MATSA. Our results indicate that MATSA is a glycoprotein and 2B9 recognizes an antigenic site in the protein moiety of MATSA. MATSA was solubilized from MSB I‐do. 18 cells by treatment with 0.5% Nonidet P‐40, and purified by affinity chromatography coupling with 2B9 and further by ion exchange chromatography on diethylaminoethylcellulose (IECD). MATSA was eluted with 0.2 to 0.3 M KCI in IECD and the purity of MATSA was increased about 2,500‐fold. The purified MATSA was shown to have a molecular weight (Mr) of 70,000 by SDS‐PAGE. The reactivity of purified MATSA with anti‐thymus cell serum was examined. MATSA was detectable by anti‐thymus cell serum, although 2B9, which was used to purify MATSA from MSBI‐clo.18 cells, was not reactive to cells prepared from the thymus. However, MATSA was no longer detectable after the absorption of anti‐thymus cell serum by chicken bursa cells. The absorption of anti‐thymus cell serum by chicken red blood cells (RBC) had no effect on the reactivity against MATSA. These results suggest that MATSA may be a lymphocyte‐specific antigen modified during leukemogenesis by Marek's disease virus (MDV).
Characteristics of five human tumor cell lines and sublines resistant to cis‐diamminedichloroplatinum(II)
Authors: Teicher Beverly A. , Holden Sylvia A. , Herman Terence S. , Sotomayor Enrique Alvarez , Khandekar Vrinda , Rosbe Kristina W. , Brann Terrence W. , Korbut Timothy T. , Iii Emil Frei
Published: February 2007

In order to study the mechanisms responsible for resistance to CDDP, 5 human tumor cell lines were made resistant to CDDP by repeated in vitro exposures. After cloning it was found that the cell lines developed were between 3.3‐fold and 17‐fold more resistant to CDDP than the parental cell lines at the IC90. These lines were also resistant to carboplatin and tetraplatin; however, resistance to tetraplatin was lower than to the other platinum complexes. Sensitivity was also assessed to Adria, MTX, 5‐FU, chlorambucil, 4‐HC, 4‐HIF, BCNU, Thiotepa, HN2, Mito C and L‐PAM, and no consistent cross‐resistance was observed. As compared with the parental lines, non‐protein sulfhydryl content was elevated in 3 resistant lines, and protein sulfhydryl was elevated in all 5 lines, as was glutathione‐S‐transferase activity. Measurements of platinum in whole cells and nuclei after exposure of the cultures to 25 μM CDDP for either 1 or 6 hr showed that nuclear levels reflected those in whole cells and that, per mg protein, platinum levels were lower in resistant cells at both time points. Formation of DNA cross‐links, determined by alkaline elution, was lower in resistant cell lines than in parental cell lines, but did not correlate with the absolute cell kill observed. These results indicate that cellular resistance to CDDP often involves decreases in drug accumulation and increases in protein sulfhydryl content. Possible strategies for overcoming these mechanisms are discussed.
Treatment of leukemia L1210 and P388 by arabinosylcytosine‐polysaccharide conjugates
Authors: Novotný Ladislav , Balážová Eliška , šandula Jozef , Ujházy Viliam , Kéry Vladimír
Published: February 2007

Conjugates of 1‐β‐D‐arabinofuranosylcytosine (araC) with two polysaccharides such as polygalacturonic acid (PGA) and carboxymethylated yeast β‐D‐glucan (CMG) were tested for their antileukemic activity in vitro on a L1210 cell line in suspension culture, in soft agar assay and in vivo on L1210, L1210/araC‐ and P388‐leukemia‐bearing mice. Both conjugates showed high activity in vitro in soft agar assay, compared with araC. Single administration of PGA‐araC or CMG‐araC increased the survival time 1.5 × or 1.7 ×, respectively, compared with araC in vivo in L1210‐leukemia‐bearing mice. The conjugates were not active against araC‐resistant leukemia line L1210/araC. The marked effect of both PGA‐araC and CMG‐araC against leukemia L1210 and P388 is probably due to the prolonged release of free araC from conjugates caused by hydrolysis.
Assessment of major carcinogens and alkaloids in the tobacco and mainstream smoke of ussr cigarettes
Authors: Djordjevic Mirjana V. , Sigountos Claire W. , Hoffmann Dietrich , Brunnemann Klaus D. , Kagan Mark R. , Bush Lowell P. , Safaev Radjab D. , Belitsky Genady A. , Zaridze David
Published: February 2007

Tobacco and mainstream smoke of USSR cigarettes were analyzed for carcinogens. The pH values of suspensions of the tobacco (5.4‐5.6) and the nitrate content of the tobaccos (0.4‐1.7%) were as expected for flue‐cured and sun‐cured tobaccos and mixtures thereof. The nicotine levels of the cigarette tobaccos (0.76‐0.94%) and total alkaloid content (0.85‐1.08%) were relatively low compared with tobaccos used in Western European and US cigarettes. The concentrations of tobacco‐specific N‐nitrosamines in the cigarette tobaccos were also low (N'‐nitrosonornicotine 0.36‐0.85 μg/g) compared with those in bright, oriental and blended cigarette tobaccos in Western countries (0.3‐19 μg/g). The 2 non‐filter and 4 filter cigarettes from the USSR had slow burning rates and yielded 14.0‐16.7 puffs/cigarette, while puff yields for commercial cigarettes in Western countries average ≤11 puffs/cigarette. Consequently, tar and benzo(a)pyrene yields in the smoke of all cigarettes as well as nitrosamine yields were high, especially in the smoke of the filter cigarettes. It appears that an increase in the burning rates of these cigarettes should lead to lower smoke yields.
Cytotoxic potential despite impaired activation pathways in T lymphocytes infiltrating nasopharyngeal carcinoma
Authors: Ferradini L. , Miescher S. , Stoeck M. , Busson P. , Barras C. , Cerf‐Bensussan N. , Lipinski M. , Von Fliedner V. , Tursz T.
Published: February 2007

Nasopharyngeal carcinoma (NPC) is an epithelial tumor consistently associated with EBV. The histological picture is characterized by a strikingly abundant lymphocytic infiltrate. Furthermore, the epithelial tumor cells present several immunological characteristics which suggest an important role for tumor‐infiltrating lymphocytes (TIL) in the biology of this tumor. The present study reports the phenotypic and functional characterization of TIL from NPC obtained after enzymatic digestion of 15 NPC biopsies. Flow cytometric analysis of TIL suspensions indicated that most TIL were mature CD3+ T lymphocytes (mean = 60%) with a variable CD4/CD8 ratio. Most TIL were TCR α/β‐positive (mean = 55%) and only a few TCR γ‐δ‐positive cells could be identified. A small percentage (mean = 9%) displayed an activated phenotype (CD25+, HLA class II+ ). Using limiting dilution analysis, we found that the average frequency of proliferative T‐lymphocyte precursors (PTL‐P) is lower among TIL (1/40) than in autologous (1/7) or normal PBL (1/1.4). Moreover, sorting experiments have shown that this defect is significantly more pronounced in the CD8+ than in the CD4 TIL subset. Accordingly, the TCR and the CD2‐mediated antigen‐independent pathways of activation were impaired. Different types of cytotoxic precursor could be detected. These included lectin‐dependent cell cytotoxicity (LDCC) and NK‐like or lymphokine‐activated killer (LAK) activity. Interestingly, some TIL from NPC were able to lyse an NPC tumor (CIS) maintained in nude mice. Thus, despite impaired activation pathways, the cytolytic potential of proliferating TIL in NPC is preserved.

This work was supported by the Institut Gustave Roussy, Villejuif, France. We would like to thank Aimée Pèle and Martine van Overloop for secretarial assistance.
Is the incidence of colorectal cancer related to reproduction? A Prospective study of 63,000 women
Authors: Kvåle Gunnar , Heuch Ivar
Published: February 2007

Relationships between reproductive factors and risk of colorectal cancer were examined in a population‐based prospective study in Norway. Available for analysis were 831 cases (581 colon cancer, 250 rectal cancer) diagnosed in a cohort of 63,090 women, surveyed in 1956‐1959 and followed through 1980. Overall, the analyses showed no strong effects of reproductive factors. In particular, high parity was not associated with reduced risk, and late age at first or last birth was not associated with an increased overall risk. However, in age‐specific analyses of colon cancer, adverse effects of late age at first birth and late age at last birth were observed in women with cancer diagnosed before the age of 60. A non‐significant overall excess risk in parous compared to nulliparous women was strongest for women with a cancer diagnosis before the age of 50 years. Having had many abortions was associated with increased risk for all sub‐sites. Neither age at menarche nor age at menopause were related to risk of colorectal cancer. The results suggest that reproductive factors, which are of importance in the etiology of cancer of the breast and genital organs in women, are not similarly related to risk of colorectal cancer.
Cytotoxic and cytostatic effects of arecoline and sodium nitrite on human cells in vitro
Authors: Wary K. K. , Sharan R. N.
Published: February 2007

Arecoline, an active alkaloid of Areca catechu L., and sodium nitrite, a food additive, are highly cytotoxic and cytostatic on the Hep 2 cell line when administered in an acidic environment (pH 4.2) in the presence of S‐9 mixture. Hep 2 cells (106) were treated with either 0.145, 0.725 or 1.449 mM sodium nitrite or 0.042,0.085 or 0.339 mM arecoline or sodium nitrite (0.145 mM) plus varied concentrations of arecoline (0.042, 0.085 or 0.339 mM). Their effects were additive in nature. Hep 2 cells exposed to this combination showed reduced cell survival, and lower rates of DNA and protein syntheses. Involvement of N‐nitroso derivatives of arecoline is suggested to explain the results. On the basis of these studies, we speculate that N‐nitroso compounds derived from arecoline can (weakly) interact with DNA.
Schedule‐dependent in vitro combination effects of methotrexate and 5‐fluorouracil in human tumor cell lines
Authors: Tsai Chun‐Ming , Gazdar Adi F. , Perng Reury‐Perng , Kramer Barnett S.
Published: February 2007

Because of conflicting reports of clinical synergy, we used the tetrazolium‐based colorimetric (MTT) assay to test in vitro combination effects of methotrexate (MTX) plus 5‐fluorouracil (FUra) in 4 schedules on 2 human non‐small‐cell lung cancer cell lines (adenocarcinoma, NCI‐H23; bronchioalveolar‐cell carcinoma, NCI‐H358), and I human colorectal adenocarcinoma cell line (SNU‐CI). The complete 3 dimensional set of isoboies in the dose range under study was generated by a microcomputer‐based method. We found that the combination effects of 8‐hr sequential FUra‐MTX, simultaneous administration of MTX‐FUra, and 8‐hr sequential MTX‐FUra were clearly antagonistic for all 3 cell lines. In contrast, the combination cytotoxic effects of 24‐hr sequential MTX‐FUra were much more active. Our in vitro model thus clearly shows that MTX‐FUra interactions are highly schedule‐dependent. This provides a rational basis for testing sequential MTX‐FUra with a longer administration interval than usually employed clinically.
A correlation between GM‐CSF gene expression and metastases in murine tumors
Authors: Takeda Kazuyoshi , Hatakeyama Kohki , Tsuchiya Yoshinori , Rikiishi Hidemi , Kumagai Katsuo
Published: February 2007

Using 14 transplantable murine tumors, we investigated a possible correlation between their ability to produce the cytokine GM‐CSF and the spontaneous metastatic potential when mice were subcutaneously inoculated. The following results were obtained: (1) seven tumors, which produced severe pulmonary metastases and metastatic swelling of lymph nodes, exhibited the ability to produce GM‐CSF activity in culture. The cell population analysis revealed that the cells producing GM‐CSF were tumor cells themselves, but that contaminating macrophages/granulocytes and T lymphocytes did not produce GM‐CSF. The mRNA for GM‐CSF was also found in all of these highly metastatic tumors tested. In mice inoculated with a highly metastatic tumor, the GM‐CSF mRNA was also found in lungs; (2) in 3 other tumors, which produced histological but not macroscopical pulmonary metastases, no GM‐CSF activity could be detected in the culture fluids. GM‐CSF mRNA was, however, detected in the tumor cells in the presence of an mRNA‐stabilizing agent, cycloheximide, suggesting the possibility that the tumor cells of this type were transcribing GM‐CSF gene, and secreting it in undetectable levels; (3) in culture of the 4 remaining poorly or non‐metastatic tumors, neither CSF activity nor GM‐CSF mRNA could be detected even in the presence of cycloheximide. GM‐CSF mRNA was also not found in lungs of tumor‐bearing mice. Our results indicate that there may be a correlation between GM‐CSF gene expression in tumor cells and spontaneous metastases.
Identification and characterization of a colon‐cancer‐associated antigen expressed on leukemia‐lymphoma cell lines
Authors: Chimori Y. , Minowada J.
Published: February 2007

A total of 72 human leukemia‐lymphoma cell lines were studied for reactivity with the monoclonal antibody (MAb) A7, an anti‐human colon‐cancer‐cell‐associated antigen reagent, by indirect membrane immunofluorescence. Nine of the 72 cell lines expressed the antigen recognized by A7 MAb. Five of the 34 T‐cell lines, 2 of the 21 B‐cell lines, and 2 of the 3 non‐lymphoid‐non‐myeloid cell lines were reactive with A7 MAb. By means of SDS‐PAGE and immunoblotting, the antigens isolated from both colon cancer cell lines (WiDr, SW1 116 and LoVo) and leukemia cell lines (A3/KAWAKAMI, H9, RPM1 8226 and SPI‐801) showed an identical MW of 42‐43 kOa. The non‐glycosylated antigen recognized by A7 MAb, which was expressed on both the colon cancer line (SW1 116) and the leukemia line (H9) in the presence of tunicamycin, also showed an identical MW of 36 kDa. However, the quantity of the antigen in the leukemia cells was significantly lower than in the colon cancer cells. Although expression of this colon‐cancer‐associated antigen in the non‐colon cancer cells is real, the significant expression of this antigen in colon‐cancer cells makes it useful for clinical monitoring of colon cancer patients.
Tumor‐cell lysis by in‐situ‐activated human peripheral‐blood mononuclear cells
Authors: Smans Karine A. , Hoylaerts Marc F. , Hendrickx Herman F. , Goergen Martine J. , De Broe Marc E.
Published: February 2007

A heteroconjugate (HC) was synthesized between OKT3 and monoclonal antibody (MAb) 7E8, which specifically reacts with the tumor marker placental alkaline phosphatase (PLAP). Similarly to OKT3, in vitro, the HC induced a dose‐dependent proliferation response of human peripheral‐blood mononuclear cells (PBMCs) and, in concert with rlL‐2, it progressively activated T cells over a 4‐day period. In co‐cultures of continuously activated PBMCs and M04 tumor cells (non‐MHC‐restricted mouse fibroblasts transfected with the cDNA for PLAP), the HC (25 ng/ml), again acting in concert with rlL‐2, induced specific lysis of the MO4 cells. This process occurred progressively over 2 to 3 days and was monitored from the release in the supernatant fluid of cellular 3H‐L‐leucine, but also from analyses involving the remaining non‐lysed cancer cells, i.e., by estimates of their protein content, by measurements of their viability, and most accurately by determinations of their PLAP content. Antibody 7E8 by itself induced a weak tumor‐cell lysis (ADCC), potentiated by the addition of rlL‐2. However, after 7 days of PBMC‐preactivation with the HC and rlL‐2, antibody 7E8 no longer mediated any ADCC, whereas the HC‐dependent lysis was further potentiated. The observed proliferation of T cells and development of cytotoxicity at low concentrations of HC and rlL‐2 support the idea that a moderate but continuous T‐cell activation combined with T‐cell targeting is sufficient for the induction of progressive and efficient tumor‐cell lysis.
Binding of epidermal growth factor‐dextran conjugates to cultured glioma cells
Authors: Andersson Annelie , Holmberg Anders , Carlsson Jörgen , Carlsson Jan , Pontén Jan , Westermark Bengt
Published: February 2007

Some gliomas, melanomas and squamous carcinomas have large numbers of EGF receptors which could, in these cases, be used for targeting with toxic agents. We investigated whether EGF could be conjugated to dextran, which is a suitable carrier for toxic agents, without losing its ability to bind to the receptor. Dextran of 20 kDa molecular weight was activated with l‐cyano‐4‐dimethylamino pyridinium tetrafluoroborate (CDAP) yielding highly active pyridinium‐isourea derivatives. EGF was coupled to the activated dextran through the amino terminus and glycine was added to block residual activity. The EGF‐dextran conjugate was, after purification on Sephadex G25 and Sephacryl 200 columns, tested for its receptor binding properties on human malignant glioma, U343MgaC12:6, cells. The conjugate inhibited binding of l25I‐EGF in a competitive assay, showing that the binding was receptor‐specific. Dextran conjugated with glycine, without EGF, had no inhibitory effect. The conjugate was radio‐labelled either on the EGF part with 125I or on the dextran part with 3H‐glycine, and the internalization patterns were compared to the internalization of 125I‐EGF. The radioactivity of the conjugates remained cell‐associated for more than 20 hr, regardless of whether the radioactivity was on the EGF or on the dextran part, while the radioactivity of unconjugated EGF rapidly disappeared from the cells. Most of the cell‐associated radioactivity was, at all analysed time intervals, located intracellularly. Thus, it seems promising to use dextran, conjugated with EGF, as a carrier of, for example, toxic radioactive nuclides.
Enhancement of natural‐killer‐cell susceptibility of human breast‐cancer cells by estradiol and v‐Ha‐ras Oncogene
Authors: Screpanti I. , Felli M. P. , Toniato E. , Meco D. , Martinotti S. , Frati L. , Santoni A. , Gulino A.
Published: February 2007

Natural killer (NK) cells are putative components of the cellular immune response to transformed cells. Since both estradiol treatment and ras‐oncogene overexpression enhance tumorigenicity of hormone‐dependent breast‐cancer cells, we studied the effects of estrogen and of the activated v‐Ha‐ras oncogene on NK susceptibility of MCF‐7 human breast‐cancer cells. MCF‐7 cells were sensitive to cytolysis mediated by resting and IL2‐activated peripheral‐blood nonadherent lymphocytes. Lysis appeared to be mediated by NK cells, since it was abrogated by treatment of effector cells with α‐CD16 monoclonal antibody (MAb) plus complement (c'). Estradiol treatment of MCF‐7 cells was able to significantly increase their sensitivity to the lysis by IL2‐activated and unactivated peripheral‐blood lymphocytes, as early as 24 hr throughout 10 days of hormone treatment. Hormone‐insensitive, estrogen‐receptor‐negative breast‐cancer cells (BT20) did not change their NK susceptibility after estradiol treatment. Increased NK susceptibility was also observed in v‐Ha‐ras‐transfected and oncogene product overexpressing MCF‐7 cells (MCF‐7‐ras) with respect to cells transfected with the selectable gene marker gpt alone (MCF‐7‐gpt). Overexpression of v‐Ha‐ras appeared to be able to bypass the need for estrogen to increase NK susceptibility, since estradiol‐treated MCF‐7‐ras cells were not lysed more than untreated MCF‐7‐ras cells. The enhancement of NK susceptibility observed after both estradiol treatment and v‐Ha‐ras overexpression suggests that the hormone‐mediated and the ras‐oncogene‐mediated signalling systems share events involved in the control of tumor‐cell/host‐effector‐cell interactions.
Growth regulation of the AML‐193 leukemic cell line: Evidence for autocrine production of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), and inhibition of GM‐CSF‐dependent cell proliferation by interleukin‐1 (IL‐1) and tumor necrosis factor (tnfα)
Authors: Kindler Vincent , Shields John , Ayer Dominique , Mazzei Gonzalo J.
Published: February 2007

The human leukemic cell line AML‐193 was tested for its proliferative response to endogenously produced autocrine factors and to a variety of cytokines and colony‐stimulating factors. Cells grown in the absence of GM‐CSF incorporated tritiated thymidine, and this was partially reversed by adding neutralizing anti‐GM‐CSF antibodies to the culture medium, suggesting that it was due, at least in part, to autocrine GM‐CSF production. This was confirmed by immunopurification of a GM‐CSF‐like activity from cell supernatant of AML‐193 cells grown in serum free medium in the absence of exogenous GM‐CSF. When AML‐193 cells were cultured with GM‐CSF in combination with other cytokines, lnterleukin‐1 α and β (IL‐I α and β), lnterleukin‐3 (IL‐3), lnterleukin‐6 (IL‐6), granulocyte colony‐stimulating factor (G‐CSF) and tumor necrosis factor a (TNFα), none of them affected the concentration of GM‐CSF required to induce 50% of maximum proliferation (D50). However, the maximum proliferation induced by GM‐CSF alone was drastically decreased by IL‐I α, IL β and TNFα. Inhibition caused by exposure of the AML‐193 to IL‐I for up to 24 hr was reversible, ruling out a direct cytotoxic effect.
Ornithine decarboxylase activity and polyamine content in adenocarcinomas of human stomach and large intestine
Authors: Berdinskikh N. K. , Ignatenko N. A. , Zaletok S. P. , Ganina K. P. , Chorniy V. A.
Published: February 2007

Ornithine decarboxylase (ODC) activity and differential polyamine composition have been studied in macroscopically normal mucosa, adjacent mucosa, and neoplastic tissue of 95 patients with adenocarcinomas of stomach and large intestine. In tumors, we found increased ODC activity and polyamine content as compared with surrounding mucosa. ODC activity in macroscopically normal tissue of patients with tumors of stomach and large intestine increased as the disease progressed. An inverse relationship was observed between ODC activity in adenocarcinomas and differentiation.
The decline in the incidence of stomach cancer in sweden 1960‐1984: A birth cohort phenomenon
Authors: Hansson Lars‐Erik , Bergström Reinhold , Sparjén PÄR , Adami Hans‐Olov
Published: February 2007

Trends in the incidence of stomach cancer among 52,604 patients notified to the National Swedish Cancer Registry in 1960 through 1984 were analysed. Age‐standardized incidence rates declined throughout the period, from 47.1 to 24.6 per 105 in males and from 23.8 to 12.7 per 105 in females. Among males the decline in incidence was more pronounced in younger age strata, 35‐54 years, while in females it reached a maximum at ages 70‐74 years. In a multivariate analysis the age‐cohort model adequately represented the data and there was no reason to separate the effects of the full age‐period‐cohort model. Compared with the cohort born in 1876‐1884, the relative risk of developing stomach cancer was 0.46 (95% Cl:0.44‐0.48) in males and 0.39 (95% Cl:0.37‐0.42) in females born in 1906‐1914. This supports the view that the declining incidence is due to a change in the exposure of the population to aetiological factors of stomach cancer and not to refinement of the diagnosis and classification of abdominal tumours. The results also imply that exposure to aetiological factors early in life together with a long induction period is of importance in the development of stomach cancer.
Risk factors for penile cancer: Results from a case‐control study in china
Authors: Brinton Louise A. , Jun‐Yao Li , Shou‐De Rong , Huang Suzanne , Sheng Xiao Bin , Bai‐Gao Shi , Zhe‐Jun Zhu , Schiffman Mark H. , Dawsey Sanford
Published: February 2007

An epidemiologic study of penile cancer involving 141 cases and 150 community controls was undertaken in a high‐risk area in China. Personal interviews, as well as physical examinations among the prospectively ascertained subjects, enabled evaluation of a variety of potential risk factors. Strongly related to risk were conditions restricting the motility of the foreskin, including phimosis or paraphimosis, particularly when so severe that circumcision was used for treatment. Poor hygiene practices also appeared to increase risk, particularly as evidenced by detection of smegma on physical examination, although it was difficult to decipher whether this association was etiologic or merely a consequence of disease. A sexual relationship outside of marriage was associated with a RR of 1.7, and appeared to be a more important discriminator than number of lifetime sexual partners. Risk was increased among subjects reporting previous genital conditions, particularly sexually transmitted diseases, and physical examinations revealed the appearance of genital warts among 13 cases vs. I control. Interviews with wives of study subjects failed to provide evidence of a “female factor” in the etiology of penile cancer. This study supports the need for further evaluation of the role of hygiene and sexually transmitted agents in the etiology of penile cancer.
Scid mouse model of epstein‐barr virus‐induced lymphomagenesis of immunodeficient humans
Authors: Purtilo David T. , Falk Kerstin , Pirruccello Samuel J. , Nakamine Hirokazu , Kleveland Kimberly , Davis Jack R. , Okano Motohiko , Taguchi Yuichi , Sanger Warren G. , Beisel Kirk W.
Published: February 2007

Immunodeficient humans are at very high risk of developing Epstein‐Barr virus (EBV)‐induced lymphomagenesis. Severe combined immunodeficient mice (SCID) have been shown to develop lymphoproliferative disease (LPD) following transfer of peripheral blood leukocytes (PBL) with latent EBV. To study mechanisms of lymphomagenesis, we compared results of engraftment of PBL from normal donors and immunodeficient donors with X‐linked lymphoproliferative disease (XLP). Graft‐versus‐host disease (GVHD) developed in 6 of 10 SCID mice 4 to 8 weeks following transfer of PBL from normal donors. In contrast, none of 9 mice engrafted with PBL from XLP patients with T‐cell defects showed GVHD. LPD developed in mice regardless of the immune competence of the donors. The expression of EBV‐encoded proteins, results of immunophenotyping, and karyotyping of the LPD lesions revealed lethal oligoclonal LPD owing to transfer of latent EBV in B cells in mice engrafted with PBL from seropositive donors. Polyclonal LPD developed in mice engrafted with PBL from seronegative patients which were infected with B95‐8 virus 6 weeks after transfer of the cells. This model is useful for investigating mechanisms of EBV‐induced LPD in immunodeficient patients.
Isolation and characterization of a serologically defined tumour membrane antigen from a chemically‐induced rat sarcoma, HSN
Authors: Léger Olivier , Dunn Pamela L. , Coles B. , Dean C. J.
Published: February 2007

Polypeptides containing the tumour antigenic determinant present on the external domain of a membrane antigen of the 3‐4 benzpyrene‐induced rat fibrosarcoma HSN have been isolated and purified. Following cleavage from intact cells with trypsin, the peptides were purified by immunoaffinity chromatography and SDS‐PAGE. Three polypeptides of molecular weight 120, 45 and 42 kDa were obtained that bound the specific rat monoclonal antibody (MAb) 11/160 in Western blots. Chemical analyses of the 45‐kDa fragment yielded the following sequence:

All 14 of the syngeneic MAbs prepared from rats bearing the HSN tumour competed with each other to bind to the specific antigen. Western blots of the purified tryptic fragments probed with 125I‐Ab, revealed that, while some of the MAbs (11/160, ALN/12/17 and ALN/9/94) recognized a sequential determinant, others (ALN/11/53, ALN/16/53, and AL/3/12) bound to a conformational epitope. It is concluded that the Ab, bind to distinct but overlapping epitopes.
Recombinant interferon alpha and gamma modulate the invasive potential of human melanoma in vitro
Authors: Hujanen E. S. , Turpeenniemi‐Hujanen T.
Published: February 2007

We have studied the effects of interferons (IFNs) on the attachment, collagenase IV activity, chemotactic migration and in vitro invasion of human melanoma (A2058) cells treated for various time periods with human recombinant interferon alpha (hrIFN‐α) or gamma (hrIFN‐γ). The cells treated with hrIFN alpha for a short time period attached more readily to purified basement membrane components, type IV collagen and laminin, than control cells. The stimulating effect of hrIFN gamma on the attachment was seen, however, when the cells were treated for a longer period of time (3 days) with this drug. The short‐term treatment with hrIFN alpha also enhanced the in vitro invasion of cells through a reconstituted basement membrane compared to findings with untreated control cells. Pre‐treatment of 3 days or more was, however, needed for hrIFN gamma to promote the invasion of A2058 cells. Both IFNs increased the secretion of basement membrane (type IV) collagen degrading metal‐loproteinase (collagenase IV) activity from human melanoma cells. Further, chemotaxis, i.e., directed migration of A2058 cells to laminin, was enhanced by both IFNs. In contrast, the attachment, collagenase IV activity, chemotaxis, and in vitro invasion were markedly inhibited when the cells were treated for an extended time period (7 days) with the IFNs. Interferons also inhibited cell proliferation after 4 days of exposure. These results suggest that time of treatment with interferons modulates the invasive capacity of human melanoma cells in vitro, causing initially a transient enhancement of invasion followed by an inhibition of invasive propensity after extended exposure to these drugs, and that different biochemical steps required for successful invasion are regulated in parallel by interferons alpha and gamma.
Tumor regression and induction of anti‐tumor immunity by local chemotherapy of guinea‐pigs bearing a line‐10 hepatocarcinoma
Authors: Claessen Anke M. E. , Valster Henriëtte , Bril Herman , Steerenberg Peter A. , Tan Ben T. G. , Scheper Rik J.
Published: February 2007

Local administration of a low dosage of the active cyclophosphamide derivative 4‐hydroperoxy‐cyclophosphamide (4‐HPCY) at the site of antigenic stimulation strongly enhances T‐cell‐mediated immune responses in both mice and guinea‐pigs. Such immunopotentiation is related to functional elimination of suppressor cells from the draining lymph nodes. In the present study we examined the potential immunotherapeutic effects of local cytostatic drug administration in strain‐2 guinea‐pigs bearing a line‐10 hepatocarcinoma. This tumor, when injected intradermally (106 cells) metastasizes within 7 days into the draining lymph node and untreated animals die within 60‐80 days from metastatic growth. In sensitization experiments, using irradiated line‐10 tumor cells, potentiation of delayed‐type hypersensitivity reactivity was observed with local administration of low dosages of 4‐HPCY. Intralesional treatment with increasing dosages of 4‐HPCY, when started 7 days after tumor‐cell inoculation and continued for 3 weeks, resulted in a dose‐dependent regression of the primary tumor. Cure rates of up to 75% were achieved. All cured animals showed strong delayed‐type hypersensitivity reactivity towards line‐10 cells and were resistant to a rechallenge with 106 line‐10 tumor cells. When treatment was started at a very late stage of the disease (day 14) only a small number of animals were cured. However, when local chemotherapy was preceded by one (non‐curative) systemic dose of cyclophosphamide, a 57% cure rate was obtained. Again, all cured animals showed strong delayed‐type hypersensitivity reactivity and protective immunity to line‐10 tumor cells. Tumor immunity was transferable to naive recipients with immune spleen cells and was T‐cell‐dependent. Other cytostatic drugs, selected for local immunopotentiating capacity, notably etoposide (VP16) and cis‐platinum (cis‐Pt) were similarly effective in the local chemotherapy protocol.
Combination of dietary factors in relation to breast‐cancer occurrence
Authors: Van T Veer Pieter , Van Leer Edith M. , Rietdijk Annelies , Kok Frans J. , Schouten Evert G. , Hermus Rudolph J. J. , Sturmans Ferd
Published: February 2007

Combinations of dietary factors were studied in relation to breast‐cancer occurrence among 133 breast cancer cases and 289 population controls in The Netherlands. Dietary factors were classified according to their possible mechanism of action, i.e., relating either to the intestinal microflora (total fat, fiber, fermented milk products) or to the anti‐oxidant hypothesis (beta‐carotene, selenium and polyunsaturated fatty acids). From 6 interactions evaluated, the combination of high fiber intake and high intake of fermented milk products was the only one suggesting synergistic protection (age‐and‐fat‐adjusted OR for interaction = 0.48, 95% confidence interval (Cl) = 0.21 − 1.13). In order to estimate the extent to which the above dietary factors together might be related to breast cancer, subjects with a supposedly favorable dietary pattern (low fat intake, high fiber intake, high intake of fermented milk products; high intake of beta‐carotene and selenium, low intake of polyunsaturated fatty acids) were compared with subjects with an unfavorable dietary pattern. This resulted in an age‐adjusted odds ratio of 0.40 (95% Cl = 0.14–1.15), which was largely attributable to the combination of low intake of fat and high intake of fermented milk products and fiber (age‐adjusted OR = 0.33, 95% Cl = 0.15 − 0.73). The other factors did not appreciably affect the odds ratio. These analyses show in a quantitative way that a dietary pattern which combines low intake of fat and high intake of fiber and fermented milk products might provide substantial protection against breast cancer.
Oligoclonal T lymphocytes infiltrating human lung cancer tissues
Authors: Yoshino Ichiro , Yano Tokujiro , Yoshikai Yasunobu , Murata Mitsuhiro , Sugimachi Keizo , Kimura Genki , Nomoto Kikuo
Published: February 2007

To clarify the nature of tumor‐infiltrating lymphocytes (TILs), we investigated the possible clonality of the T cells in TILs freshly isolated from human primary lung cancer tissues by assessing the rearrangement pattern of the T‐cell receptor (TCR) gene P locus using Southern blotting. First, in phenotypic analysis, TILs represented different populations among corresponding peripheral blood lymphocytes (PBLs) with an increased proportion of CD20+ (B) cells as well as a decreased proportion of CD 16+ (natural killer) cells, and a variable CD4/ CD8 ratio. Considering the central role of T cells in immune responses, we analyzed TCR β gene rearrangement patterns in TILs and corresponding PBLs from 12 patients. In 10 of the 12 cases, TILs showed one or more TCR gene rearrangement bands with a predominance of the C.β2 gene, in which 2 types of common rearranged band were observed among the cases with different clinical profiles in terms of histological types and disease stage, with bands at about 9.5 kb in 7 and at 11.5 kb in 8 patients. On the other hand, predominant rearranged bands were hardly detected in corresponding PBLs except in 2 cases. From these results, we conclude that TILs in lung cancer tissues frequently contain oligoclonal T‐cell populations, which were probably sensitized by relatively common antigens at the tumor sites.
A phase‐i study of repeated therapy with radiolabelled antibody to carcinoembryonic antigen using intermittent or continuous administration of cyclosporin a to suppress the immune response
Authors: Ledermann J. A. , Begent R. H. J. , Massof C. , Kelly A. M. B. , Adam T. , Bagshawe K. D.
Published: February 2007

The anti‐mouse antibody response was examined in patients receiving repeated i.v. therapy with radiolabeled mouse monoclonal antibody (MAb) to carcinoembryonic antigen (CEA): 131I anti‐CEA was given approximately every 2 weeks with cyclosporin A, to suppress the anti‐mouse antibody response. Two schedules of cyclosporin A–intermittent therapy for 6 days with each course of anti‐CEA and continuous therapy–were compared. Suppression of the immune response in the intermittent high‐dose (3 patients) and continuous low‐dose groups (4 patients) was equivalent, but the latter regimen was less toxic. Repeated therapy led to the formation of small amounts of anti‐mouse antibody, but provided that cyclosporin A was continued it did not prevent further therapy or lead to an increase in the rate of clearance of anti‐CEA from blood. Without cyclosporin A no more than 2 courses of antibody therapy could be given. Patients received up to 4 doses of ml anti‐CEA. The nadir platelet count was related to the half‐life of 131I anti‐CEA in blood. Thrombocytopenia limited the amount of 131I anti‐CEA that could be given and determined the interval between treatments. Effective suppression of the anti‐antibody response is possible and this study has determined that myelosuppression is the principal obstacle to repeated therapy with radiolabeled antibody.
Fetal myofibroblast‐like cells isolated from post‐radiation fibrosis in human breast cancer
Authors: Brouty‐Boyé D. , Raux H. , Azzarone B. , Tamboise A. , Tamboise E. , BéRanger S. , Magnien V. , Pihan I. , Zardi L. , Israël L.
Published: February 2007

Cells were isolated from post‐radiation fibrosis biopsies of patients with recurrent breast carcinoma. These cells were identified as fibroblasts and compared with fibroblasts from normal breast tissues for their proliferative activities, chromosome number and for the presence of various components of the extracellular matrix and cytoskeleton. The proliferative activity of the fibrosis‐derived fibroblasts did not significantly differ from that of normal breast fibroblasts. Both cell types required serum to grow and did not form colonies in soft agar. Cells from 2 of the 3 fibroses analyzed displayed aneuploid karyotypes with multiple structural abnormalities. All of the fibroblastic cells produced types I, III and V collagen, fibronectin and vimentin. However, in contrast to normal breast fibroblasts, fibrosis‐derived cells produced high amounts of oncofetal fibronectin. In addition, fibrosis fibroblasts also expressed the a‐actin isoform which is specific for smooth‐muscle cells. These results suggest that post‐radiation fibrosis in malignant breast contains atypical fibroblasts with fetal and myofibroblastic characteristics.
The association between alcohol and breast cancer risk: Evidence from the combined analysis of six dietary case‐control studies
Authors: Howe Geoffrey , Rohan Thomas , Decarli Adriano , Iscovich José , Kaldor John , Katsouyanni Klea , Marubini Ettore , Miller Anthony , Riboli Elio , Toniolo Paolo , Trichopoulos Dimitrios
Published: February 2007

Data from 1,575 cases and 1,974 controls enrolled in 6 previously conducted case‐control studies of diet and breast cancer have been analysed with respect to alcohol intake. There appears to be an absence of any association between consumption of up to 40 g of alcohol per day and risk of breast cancer, and a highly statistically significant and consistent elevated risk of breast cancer for drinkers of 40 g or more of alcohol per day, for whom the relative risk, as compared with that of non‐drinkers, is 1.69 (95% confidence interval 1.19 to 2.40). This association is not due to confounding by a number of diet‐related factors, including total calories, fat, fibre and vitamin C.
Loss of a tumor suppressor function during neoplastic progression of epithelial cells in vitro
Authors: Knowles Margaret A. , Eydmann Marian E.
Published: February 2007

In vitro transformation of rat urothelial cells is a multi‐step process. We have used cell fusion to analyse the role of recessive events during in vitro progression of an immortal urothelial cell line. Somatic cell hybrids were made between the transformed cell line RM2T and a series of immortal urothelial cell lines, including the progenitor line RM2AD, from which RM2T was isolated. The ability to produce colonies in soft agar (anchorage independence) was used as an in vitro marker of transformation, and a series of 10 hybrid clones and 4 mass populations of hybrids were assessed for suppression of this phenotype. Hybrids between early‐passage (< passage 35, anchorage‐dependent) RM2AD cells and late‐passage (>passage 35, anchorage‐independent) RM2T cells, showed suppression of anchorage independence when tested early after fusion (4/4 mass populations, 7/10 clones). This indicates that in vitro progression of this cell line is associated with loss of a function which can suppress growth in soft agar. Fusions between anchorage‐independent RM2T cells and a series of other anchorage‐dependent immortal urothelial cell lines generated hybrids which showed no suppression of anchorage independence, indicating that these anchorage‐dependent cells have lost the suppressor function identified in RM2AD. Our results indicate that loss of a suppressor function can contribute to urothelial transformation in vitro and that clonal populations of immortal cells, at apparently the same stage of transformation, differ in their ability to suppress anchorage independence of the cell line RM2T. These differences provide the basis for suppressor‐gene cloning experiments based on gene transfer.
Different drug sensitivity in two neuroblastoma cell lines established from the same patient before and after chemotherapy
Authors: Kuroda Hiroshi , Sugimoto Tohru , Ueda Kazumitsu , Tsuchida Shigeki , Horii Yoshihiro , Inazawa Johji , Sato Kiyomi , Sawada Tadashi
Published: February 2007

Drug resistance is one of the major impediments to the treatment of advanced neuroblastoma. Two neuroblastoma cell lines established from the same patient before (KP‐N‐AY) and after (KP‐N‐AYR) chemotherapy are described. Both cell lines were established from bone‐marrow metastases of a 2½‐year‐old patient with stage IV neuroblastoma. Chromosomal analysis, catecholamine assessment and the surface membrane phenotype of these cell lines confirmed that the tumors were of neuroblastoma origin. Compared with the KP‐N‐AY cell line, the KP‐N‐AYR line had decreased N‐myc amplification but increased N‐myc expression. An in vitro sensitivity test using a clonogenic assay showed the KP‐N‐AYR cell line to be 3.0‐fold resistant to adriamycin and 2.7‐fold resistant to cis‐platinum as compared with the KP‐N‐AY cell line. The expression of the multi‐drug‐resistance gene (MDRI) was not observed in either cell line by the ribonuclease protection assay. The KP‐N‐AY cell line revealed only faint MDRI RNA by the polymerase chain reaction, whereas the KP‐N‐AYR cell line had no expression of the MDRI gene. The level of glutathione‐S‐transferase‐π was significantly higher in the KP‐N‐AYR cell line than in the KP‐N‐AY cell line. These findings suggest that the development of clinical drug resistance may be associated with the enhanced glutathione‐S‐transferase‐π activity but not with MDRI gene expression.
Changes in the ganglioside composition of human neuroblastoma cells under different growth conditions
Authors: Sasaki Hiroshi , Momoi Torn , Yamanaka Chutaro , Yorifuji Tohru , Kaji Masayuki , Mikawa Haruki
Published: February 2007

The ganglioside composition of human neuroblastoma cells (LA‐N‐1 and LA‐N‐5) was studied in samples obtained from (1) original cells in tissue cultures, (2) tumors grown in nude mice inoculated with original cells and (3) cells in tissue cultures re‐established from the mouse tumors. The amounts of „a”︁ pathway gangliosides (Gm2, G m1 and GD1a) and those of the „b”︁ pathway (GD3, GD2, GD1b and GT1b) differed according to the culture conditions. The „b”︁ pathway gangliosides were markedly increased in the tumors grown in nude mice. In contrast, the „a”︁ pathway gangliosides were abundant in cultures of both original and re‐established cells. We also measured the enzymatic activities of UDP‐N‐acetylgalac‐tosamine: GM3, N‐acetylgalactosaminyl transferase (EC 2.4.1.92) and of CMP‐N‐acetylneuraminic acid: GM3 sialyl transferase (EC 2.4.99.8) in neuroblastoma cells cultured under these conditions. These enzymes are thought to be the key enzymes involved in the synthesis of the „a”︁ and „b”︁ pathway gangliosides. Though there was no significant difference in the activity of N‐acetylgalactosaminyl transferase between original cells and tumors in nude mice, re‐established cells showed a definitely higher activity (3.5 times higher than in the original cells). On the other hand, tumors grown in nude mice had a markedly higher activity of sialyl transferase than that of original cells or re‐established cells. These findings suggest that the culture conditions and/or the type of cell growth play some role in the synthesis and expression of gangliosides in neuroblastoma cells.
A comparative study on the xenotransplantability of human solid tumors into mice with different genetic immune deficiencies
Authors: Zietman A. L. , Sugiyama E. , Ramsay J. R. , Silobrcic V. , Yeh E. T. H. , Sedlacek R. S. , Suit H. D.
Published: February 2007

These experiments set out to assess the role of NK and B cells in the resistance of nude mice to human tumor xenotransplantation. The transplantability of 9 fresh and 8 cultured human tumors was compared in 2 strains of mice with different genetic immune deficiencies: athymic NCr/Sed (nu/nu) nude mice, and nude‐beige‐xid (N:NIH‐nu‐bg‐xid/Sed mice). Flow cytometric studies showed both strains to be deficient in Thy. 1.2 (T) cells and unresponsive to stimulation by Concanavalin A (Con A) or direct T‐cell‐receptor triggering with anti‐CD3. The number of B cells was similar in the 2 strains, but the response to lipopolysaccharide (LPS) was markedly reduced in the nude‐beige‐xid animals. The number of asialoGM1,‐positive cells (predominantly NK) detected by flow cytometry was also reduced in the nude‐beige‐xid mice. The transplantability of the human tumors was found to be equivalent in the 2 strains. Quantitative cell‐transplantation assays performed for 2 of the tumor cell lines did not reveal any subtle transplantation advantage for the more broadly immune‐deficient animals. No evidence could, therefore, be found to suggest that NK or B cells were major determinants of human tumor xenotransplantability in these strains of mice.