bluuebunny/crossref_metadata_2025 huggingface dataset:
Growth‐inhibitory effect of quercetin and presence of type‐II estrogen‐binding sites in human colon‐cancer cell lines and primary colorectal tumors
Authors: Ranelletti Franco O. , Ricci Riccardo , Larocca Luigi M. , Maggiano Nicola , Capelli Arnaldo , Scambia Giovanni , Benedetti‐Panici Pierluigi , Mancuso Salvatore , Rumi Carlo , Piantelli Mauro
Published: February 2007

We studied the effect of quercetin (Q) on the proliferation of HT‐29, WiDr, COLO 201, and LS‐174T human colon cancer cell lines. Q, between 10 nM and 10 μM, exerted a dose‐dependent, reversible inhibition of cell proliferation. Cell‐cycle analysis revealed that the growth‐inhibitory effect of Q was due to a blocking action in the G0/G1 phase. Using a whole‐cell assay with 17β‐[3H]‐estradiol as tracer, we demonstrated that all these cell lines contain type‐II estrogen‐binding sites (type‐II EBS). By using Q and other chemically related flavonols (3,7‐4′‐trimethox‐ yquercetin, 3,7,3,4′‐tetramethoxyquercetin, kaempferol, morin, and rutin), we observed that the affinities of these compounds for type‐II EBS are correlated with their growth‐inhibitory potential. Furthermore, the Q sensitivity of the colon cancer cell lines was correlated with the number of type‐II EBS/cell. Then Q could regulate colon cancer cell growth through a binding interaction with type‐ll EBS. This mechanism could also be active in vivo as we have observed that cytosolic type‐II EBS are present in primary colorectal cancers and that Q is effective in inhibiting the in vitro bromodeoxyuridine incorporated by neoplastic cells in these cancers.
Enhanced insulin‐receptor tyrosine kinase activity associated with chromosomal translocation (1;19) in a pre‐B‐cell leukemia line
Authors: Newman Julie D. , Harrison Leonard C. , Eckardt Glenn S. , Jack Ian
Published: February 2007

The gene for the insulin receptor has been assigned to chromosome 19 near the breakpoint of the translocation t( 1; 19) which occurs in 25% of pre‐B‐cell leukemias. Insulin receptors in a pre‐B‐cell leukemia cell line (ACV) with t(1;19) were found to have 2‐fold higher affinity for insulin, 5‐fold higher basal and insulin‐stimulated p sub‐unit autophosphorylation, and 2‐fold higher basal and 4‐fold higher insulin‐stimulated β sub‐unit kinase activity on the synthetic peptide poly(Glu,Tyr), compared to receptors in a B‐cell line (ADD) with normal karyotype from the same patient. ACV cells had a novel 13‐kb receptor mRNA species and expressed a DNA polymorphism localized to the tyrosine kinase domain of the receptor gene. These findings suggest that t(l;19) in the ACV cell may result in rearrangement of the insulin receptor gene and translation of a receptor with enhanced tyrosine kinase activity.
Expression and characterization of two differentiation antigens in human stratified squamous epithelia and carcinomas
Authors: Quak Jasper J. , Schrijvers Ad H. G. J. , Brakkee Joost G. P. , Davis Hugh D. , Scheper Rik J. , Meijer Chris J. L. M. , Snow Gordon B. , van Donoen Guus A. M. S.
Published: February 2007

Using viable cells of a human squamous‐cell carcinoma (SCC) cell line as immunogen, we generated 2 monoclonal antibodies, MAbs K984 and K928, to SCC surface antigens. Immunoperoxidase staining of frozen sections of normal epidermis revealed that MAb K984 reacts with the poorly differentiated basal cells, while MAb K928 is reactive with the more highly differentiated suprabasal cells. A similar complementary reaction pattern of these antibodies was demonstrated in the majority of well‐differentiated human tumors and some moderately differentiated SCCs. In contrast, simultaneous reactivity of MAb K984 and K928 was found for the majority of cells within other well‐and moderately differentiated SCCs, as well as all poorly differentiated SCCs. Further biochemical characterization indicated that the antigen recognized by MAb K984 is similar to the one recognized by MAb SF‐25. MAb K928 recognizes a 50‐to 55‐kDa molecule under non‐reducing conditions. Antibodies with similar features to MAb K928 have not been described previously. The antigens recognized by MAbs K984 and K928 can be regarded as novel markers associated with cellular maturation in squamous epithelia. The antigen detected by MAb K984 is probably associated with the proliferating fraction in SCCs.
A monoclonal antibody capable of detecting new differentiation antigen of the stomach
Authors: Yokozaki Hiroshi , Kvo Eikai , Ochiai Atsushi , Ito Masanori , Tio Hisao , Tabuchi Junji , Tahara Eiichi
Published: February 2007

A monoclonal antibody (MAb) was raised against a gastric adenocarcinoma cell line TMK‐1 that shows parietal cell differentiation. This MAb, an IgMK, named 20D11, reacted immunohistochemically with the intracytoplasmic secretory canalicular membranes of normal parietal cells of the stomach. In the fetal stomach at 4 months of gestation, the luminal surface of fundic epithelia and developing parietal cells had MAb20D 11 immuno‐reactivity, but at 6 months of gestation only parietal cells showed immunoreaction. Out of 133 gastric carcinomas, 98 (74%) had MAb20D 11 immunoreactivity. In the well‐differentiated type, immunoreactivity was localized at the luminal surface of tumor cells and, in the poorly differentiated type, in the intracytoplasmic tubulovesicles of tumor cells. In Western‐blot analysis, the antigenic molecule had a molecular weight exceeding 300 kDa in the tumor tissues, but only a 62‐kDa immunoblot band was seen in the corresponding non‐cancerous gastric mucosa. However, there was no correlation between the expression of high‐molecular‐weight antigens and the histological types of gastric carcinoma. These results suggest that 20D11 antigen may be involved in differentiation of parietal cells in the fetal stomach and in stomach cancer.
Nucleoside diphosphate kinase/NM23 expression in breast cancer: Lack of correlation with lymph‐node metastasis
Authors: Sastre‐Garau X. , Lacombe M. L. , Jouve M. , Véron M. , Magdelénat H.
Published: February 2007

The product of the nm23‐H1 gene, reported to be a meta‐static suppressor gene, was recently identified as the nucleoside diphosphate (NDP) kinase A, and was found to be over‐expressed in several types of malignant tumors as compared with normal tissues. In order to determine whether NDP‐kinase expression serves as a marker for metastatic potential and whether hyperproliferation of neoplastic cells would correlate with expression, we analyzed NDP‐kinase levels and activity by immunohistochemical staining and by an enzymatic assay in 13 benign and 98 malignant breast‐tissue specimens. Our results confirm that NDP‐kinase expression increases in malignant cells of breast carcinomas, but cannot be considered as a biological marker of metastatic dissemination. No correlation was found between NDP‐kinase activity and S phase, taken as an index of cell proliferation. Moreover, no correlation was observed between NDP‐kinase activity and tumor size, histoprognostic index, estrogen receptors or progesterone receptors. The mechanism of over‐expression of NDP in malignant cells and its role in tumor progression remain to be determined.
Differential expression of the CD44 molecule in human brain tumours
Authors: Kuppner M. C. , Van Meir E. , Gauthier Th. , Hamou M‐F. , De Tribolet N.
Published: February 2007

Expression of the CD44 molecule was examined in a variety of human brain tumours, brain metastases and normal brain. Immunohistological staining with several CD44 antibodies demonstrated differential expression of the CD44 molecule among different brain tumour types. CD44 was strongly expressed in high‐grade gliomas and weakly expressed in meningiomas, medulloblastomas and normal brain. Northern blot analysis revealed the presence of 3 major CD44 mRNAs of 1.6, 2.2, and 5.0 kb in glioblastomas and a mRNA of 5.6 kb in meningiomas. CD44 expression was also detected by flow cytometric analysis on cultured cells derived from a variety of human brain tumours including glioblastomas and meningiomas.
Stimulation of the metastatic properties of lewis‐lung‐carcinoma cells by autologous granulocyte‐macrophage colony‐stimulating factor
Authors: Young M. Rita I. , Lozano Yvonne , Coogan Michael , Wright Mark A. , Young Melvin E. , Bagash Jamila M.
Published: February 2007

Using both polymerase‐chain‐reaction analysis and the soft‐agar colony‐forming unit assay, granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) was shown to be expressed by cloned metastatic Lewis‐lung‐carcinoma (LLC‐LN7) cells but not by non‐metastatic LLC‐C8 cells. Furthermore, the metastatic LLC‐LN7 cells were shown to respond both to autologous GM‐CSF and to exogenous recombinant GM‐CSF (rGM‐CSF). In the presence of neutralizing anti‐GM‐CSF antibodies, the metastatic LLC cells became less able to migrate or to adhere and invade through a reconstituted basement membrane. Moreover, the addition of rGM‐CSF further enhanced the capacity of the metastatic LLC cells to adhere to the reconstituted basement membrane. This stimulation of metastatic properties of the LLC cells by either autologous or exogenous GM‐CSF was associated with enhanced endogenous protein phosphorylation. Two proteins of approximately Mr 45,000 and Mr 64,000 were the dominant target proteins to be phosphorylated by the presence of GM‐CSF. These results suggest that autologous GM‐CSF may function as an autocrine stimulator of the metastatic properties of metastatic LLC cells.
Potent anti‐tumorigenic effect of tubeimoside 1 isolated from the bulb of Bolbostemma paniculatum (maxim) franquet
Authors: Yu Lijian , Ma Rundi , Wang Yongqing , Nishino Hoyoku , Takayasu Junko , He Weizhen , Chang Mei , Zhen Jie , Liu Wensheng , Fan Suxian
Published: February 2007

Tubeimoside 1, one of the new triterpenoid saponins from the bulb of Bolbostemma paniculatum (Maxim) Franquet, had an anti‐inflammatory effect on mouse ear edema induced by arachidonic acid and 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA). Furthermore, a potent anti‐tumorigenic effect of tubeimoside I was observed in 2‐stage carcinogenesis of mouse skin after oral administration as well as topical application. Thus, tubeimoside I appears to be a promising agent for cancer chemoprevention.
Malignant transformation of human fibroblasts previously immortalized with 60Co gamma rays
Authors: Mihara K. , Bai L. , Kano Y. , Miyazaki M. , Namba M.
Published: February 2007

Normal human fibroblasts obtained from a whole embryo were malignantly transformed with only 60Co gamma rays, but after extensive passage. The fibroblasts were exposed intermittently to 60Co gamma rays, 13 times, 2,800 rads in total, from the primary culture to the 40th passage level. At the 50th passage level the cells became morphologically changed, showing chromosomal abnormalities, and serial passages showed them to be immortalized. They were not tumorigenic on transplantation into nude mice, but became malignant after extensive passage; i.e., at the 547th passage level and 2,800 days after initiation of the culture. Our results indicate that several different genotypic changes are necessary for malignant transformation of human cells over long periods. No mutation at codons 12 and 61 of H‐, K‐ and N‐ras was detected in the malignant cells. Thus, this system may be useful to detect other cellular genes that may contribute to the malignant phenotypes of human fibroblasts.
Response of FR3T3 cells transformed by HA‐ras oncogene and epidermal growth factor gene to differentiation induction by N,N‐dimethylformamide
Authors: Levine Alan E. , Chakrabarty Subhas
Published: February 2007

Treatment of chemically transformed fibroblasts with N,N‐dimethylformamide (DMF) results in the restoration of a non‐transformed phenotype. In an attempt to identify more precisely the mechanisms by which DMF reverses the transformed phenotype, the effects of DMF on fibroblasts which were transformed by a single gene—specifically a synthetic epidermal growth factor (EGF) gene or the Ha‐ras oncogene—were examined. The constitutive expression of either the Ha‐ras oncogene or the EGF gene in FR3T3 fibroblasts resulted in cellular transformation. The effect of the differentiation‐inducing agent DMF on several properties of these transformed cell lines was examined. The EGF‐transfected cells were much more responsive to DMF than the ras‐transfected cells. DMF treatment of the EGF‐transfected cells resulted in the inhibition of anchorage‐independent growth, the restoration of a normal cellular morphology and growth rate in monolayer culture, and the down‐regulation of the proliferation‐associated nucleolar protein B23. DMF treatment had a much slighter effect on growth of the ras‐transfected cells in monolayer culture or under anchorage‐independent growth conditions. The high proliferation rate of the ras‐3 cells was associated with elevated expression of protein B23. The 19‐3 cells, but not the ras‐3 cells, expressed cell‐surface fibronectin. Treatment of the ras‐3 cells with DMF did not restore fibronectin expression. The binding of EGF was increased 3‐fold in the EGF‐transfected cells and decreased 20‐fold in ras‐transfected cells, but in neither case did DMF alter EGF binding. DMF treatment increased the secretion of EGF in the 2 transfected lines as well as in control cells. These results suggest that the aberrant‐growth control in the EGF‐transfected cells, but not in the ras‐transfected cells, could be modulated by DMF and that the aberrant‐growth control mechanisms were different in these 2 cell types.
Invasive and metastatic properties of MCF‐7 cells and rasH‐transfected MCF‐7 cell lines
Authors: Gelmann Edward P. , Thompson Erik W. , Sommers Connie L.
Published: February 2007

In vitro invasion and in vivo metastasis assays were performed with a panel of MCF‐7 cells transfected with isogenic constructs of mutated rasH genes. Both increased levels of rasH expression and rasH oncogene activation increased activity of derivative cell lines in in vitro invasion assays. In vivo formation of spontaneous metastases was assessed after intradermal inoculation of MCF‐7 cells in the vicinity of the mammary fat pads of ovariectomized nude mice. No metastases were seen in the absence of estradiol treatment of the mice. With estradiol supplementation of the mice both the rasH‐transfected and control transfected cell lines gave a higher incidence of metastases than parental MCF‐7 cells. Prolonged treatment of mice with exogenous estradiol (60 days vs. 21 days) resulted in more frequent metastases to liver and lung at the end of the 90‐day observation period. In contrast to activated rasH‐gene enhancement of metastatic capacity of rodent fibroblast and epithelial cell lines, there was no correlation of rasH expression with in vivo metastatic capacity of a human mammary carcinoma cell line.
Confluence‐dependent resistance in human colon cancer cells: Role of reduced drug accumulation and low intrinsic chemosensitivity of resting cells
Authors: Dimanche‐Boitrel Marie‐ThéRéSe , Pelletier Hélène , Genne Philippe , Petit Jean‐Michel , Grimellec Christian Le , Canal Pierre , Ardiet Claude , Bastian Gérard , Chauffert Bruno
Published: February 2007

In vitro sensitivity of HT29 human colon cancer cells to doxorubicin (DXR), vincristine (VCR), etoposide (VP16), cisplatin (CDDP), melphalan (L‐PAM) and 5‐fluorouracil (5FU) was markedly reduced when cell‐culture density increased. For some drugs, confluence‐dependent resistance (CDR) was partly due to decreased intracellular drug accumulation; the ratio of mean intracellular drug content of non confluent to confluent cells (NC/C) was 2.5 for DXR, 4.1 for VCR and 7.4 for VP16. Altered drug penetration with confluence could be related to decrease of plasma membrane fluidity as measured by the fluorescence polarization method. Reduction of drug intracellular accumulation was nil or weak for L‐PAM (NC/C = 1.0), CDDP (NC/C = 1.2) and 5 FU (NC/C = 1.8). Even if drug concentration was adjusted in culture medium to produce similar intracellular drug content in confluent and non confluent cells, higher intrinsic resistance of confluent cells was still evidenced for DXR and VP16 but not for VCR, the only agent without direct interaction with DNA. DXR‐ and VP16‐induced DNA breakage was also less important in confluent than in non‐confluent cells. CDR appeared closely related to an increased proportion of non‐cycling cells at confluence, as demonstrated by flow cytometry, expression of nuclear antigen recognized by Ki67 MAb and expression of topoisomerase II. CDR is probably a major factor in the poor sensitivity of colorectal adenocarcinomas to chemotherapy.
Over‐expression of p53 nuclear oncoprotein in colorectal adenomas
Authors: Pignatelli Massimo , Stamp Gordon W. H. , Kafiri Georgia , Lane David , Bodmer Walter F.
Published: February 2007

p53 is a nuclear phosphoprotein which controls normal cell growth. Normal p53 protein is undetectable by standard immunohistochemical staining and the over‐expression found in neoplastic cells correlates with the presence of point mutations of evolutionary conserved regions of the p53 gene. We examined the expression of p53 protein in a series of 36 colorectal adenomas (13 tubular, 17 tubulovillous, 6 villous) showing different degrees of dysplasia (11 mild, 19 moderate, 6 severe), 11 moderately differentíated adenocarcinomas (6 Duke's A, 4 Duke's B, 1 Duke's C) and 5 metaplastic polyps using the polyclonal antibody CM1 which recognises p53 protein in conventionally fixed and processed histological material. We found that 15 out of 36 colorectal adenomas showed p53 immunoreactivity, although in 4 positive cases (26%) the staining was very focal (<0.1% positive cells). More than 80% of severely dysplastic adenomas showed strong p53 immunoreactivity and this over‐expression was correlated with increased cell proliferative rate as detected by the proliferating‐cell‐nuclear‐antigen (PCNA) staining. p53 nuclear staining was also seen in 8 out of 11 (65%) colorectal adenocarcinomas as previously shown. Our data suggest that the p53 gene mutation, with the subsequent over‐expression of the protein, occurs in colorectal adenomas and may therefore be a fundamental genetic event underlying the dysplasia and loss of proliferative control that are characteristic of adenomas with malignant potential.
Effect of a single infusion of alendronate in malignant hypercalcaemia: Dose dependency and comparison with clodronate
Authors: Rizzoli R. , Buchs B. , Bonjour J.‐P.
Published: February 2007

Among aminohydroxybisphosphonate derivatives, atendronate (3‐amino‐1‐hydroxybutylidene‐1, 1‐bisphosphonate) has proved efficacious in diseases with increased bone resorption. However, the effective dose in the treatment of malignant hypercalcaemia is not clearly established. In 2 randomized studies, we investigated the effects of alendronate and of clodronate (dichloromethylene bisphosphonate) given as a single infusion in 82 rehydrated patients with malignant hypercalcaemia. Various doses of alendronate or clodronate soon produced a significant fall in plasma calcium (Ca), accompanied by a dose‐dependent decrease in the fasting urinary Ca/creatinine ratio, taken as a reflection of bone resorption. During the next 5 days, plasma Ca and fasting urinary Ca/creatinine ratio were lower in the group treated with alendronate than in the clodronate group. The renal handling of Ca was similar in both groups. Because of relapsing hypercalcaemia, some patients received an infusion of alendronate approximately 2 weeks after the first infusion; this normalized the urinary Ca/creatinine ratio in 44% of the cases at day 3. At that time, the plasma Ca was below 2.70 mmol/l in 33%. Our results indicate that alendronate decreased bone resorption and calcaemia in cancer patients in a dose‐dependent manner.
Autoradiographic and flow‐cytometric assessment of cell proliferation in primary colorectal cancer: Relationship to dna ploidy and clinico‐pathological features
Authors: Costa A. , Faranda A. , Scalmati A. , Quagliuolo V. , Colella G. , De Leon M. Ponz , Silvestrini R.
Published: February 2007

Two cell‐kinetic parameters, the [3H]thymidine‐labeling index [3H]dT LI and the flow‐cytometric S‐phase cell fraction (FCM‐S), and DNA ploidy were determined for a prospective series of 110 primary colorectal cancers. Aneuploidy was observed in 66% of tumors and more than one aneuploid peak was present in 12%. The frequency of aneuploid tumors was higher in rectal (80%) and left‐colon (70%) cancers than in right‐colon cancers (51%), and multiple aneuploid clones were detected more frequently in men than in women (p = 0.03) and more frequently in advanced Dukes' D‐stage patients (p = 0.08). The median [ H]dT LI value (17.4%) was similar to the FCM‐S value determined by a planimetric model (16.2%) and somewhat higher than the FMC‐S value obtained by an optimization procedure (11.2%). However, there was no significant relationship between the [3H]dT LI value and either FCM‐S value for individual tumors. Moreover, FCM‐S values were higher in aneuploid than in diploid tumors, whereas [3H]dT LI values were independent of DNA‐ploidy status; [3H]dT LI and FCM‐S were also related differently to some clinical and pathological features such as tumor site and histology. These findings suggest different biological meanings for these 2 cell‐kinetic parameters, which should not be used interchangeably.
Protein kinase C activity and expression in normal and adenomatous human pituitaries
Authors: Alvaro V. , Touraine P. H. , Vozari R. Raisman , Bai‐Grewer F. , Birman P. , Joubert (Bression) D.
Published: February 2007

Protein kinase C (PKC) activity and expression were measured in 54 adenomas (prolactin (PRL)‐, growth hormone (GH)‐and non‐secreting), I of them obtained from a patient treated with the dopamine agonist bromocriptine and 2 from patients treated with the somatostatin analog octreotide. They were also measured in normal human and rat pituitaries. Total PKC activity was measured by incorporation of 32P into histones, and PKC expression by dot blot immunoquantification using purified PKC as a standard. Both enzyme activity and expression were higher in adenomatous pituitaries than in normal human or rat pituitaries. PKC expression in GH‐secreting and non‐secreting tumors was significantly higher than that in PRL‐secreting tumors. Furthermore, it was significantly higher in invasive tumors than in non‐invasive tumors. In the 3 adenomas which were obtained from patients treated with bromocriptine or octreotide and which were used for PKC‐activity measurement, particulate‐ and soluble‐PKC activities were significantly lower than those measured in non‐treated adenomas.
Differential screening of a human pancreatic adenocarcinoma λgt11 expression library has identified increased transcription of elongation factor ef‐1α in tumour cells
Authors: Grant Anne G. , Flomen Rachel M. , Tizard Mark L. V. , Grant David A. W.
Published: February 2007

A human pancreatic adenocarcinoma λgt11 expression library was differentially screened with mRNA derived from normal and cancerous pancreatic tissues. Five clones preferentially hybridized with pancreatic adenocarcinoma mRNA. cDNA inserts from 4 of these clones were amplified by PCR, labelled with α32P and used in Northern blot analysis against mRNA prepared from a variety of tumour and normal tissues. λGER‐4 identified a pancreas‐associated mRNA (> 10 kb) with no homology with known sequences at either the nucleic or amino‐acid level. λGER‐2 identified a 1.7‐kb mRNA transcript that was over‐expressed in mRNA prepared from pancreas, colon, breast, lung and gastric tumours relative to normal tissues. Sequence analysis and restriction‐enzyme mapping showed that this clone was completely homologous with the active form of human elongation factor EF‐1α. This high level of EF‐1α‐mRNA expression in tumour tissues lends support to the increasing evidence that EF‐1α is an important regulator of the cell cycle.
Differential cytostatic activity of monocyte‐derived cytokines against human melanoma cells
Authors: Te Velde Anje A. , Kemenade Elly V. D. Wiel‐V. , Figdor Carl G.
Published: February 2007

We investigated the capacity of 3 major cytokines secreted by activated monocytes, IL‐1β, TNFα and IL‐6, to inhibit growth of melanoma tumor cells. Using neutralizing antibodies against IL‐1β, TNFα and IL‐6, we observed that the cytostatic activity against A375 melanoma cells is largely due to the presence of IL‐6 in culture supernatants of monocytes stimulated with LPS. A375 cells appeared to be extremely sensitive to monocyte‐derived cytokines, since in addition to rIL‐6 also rIL‐1β and rTNFα displayed cytostatic activity against A375 cells. We observed additive or synergistic cytostatic effects upon use of combinations of these cytokines. When 7 other melanoma cell lines and short‐term melanoma cultures were tested and compared with A375, a major difference in their sensitivity to monocyte‐derived cytokines were observed. Although 7 out of 8 melanoma cell lines were sensitive to culture supernatants of monocytes stimulated with LPS, significant differences were found when recombinant cytokines were used. The widely used A375 was the only melanoma cell line sensitive to rIL‐1β, rTNFα and rIL‐6. The growth of none of the other 7 melanoma cell cultures was significantly affected by rIL‐1β. Seven out of 8 melanoma cell cultures were sensitive to rTNFα and 3 out of 8 to rIL‐6. The results of our study indicate that the sensitivity of melanoma cell cultures for different monocyte‐derived cytokines is highly variable, and that it is questionable whether the A375 melanoma cell line, sensitive to rIL‐1β, rTNFα and rIL‐6, is representative for melanoma.
Experimental studies on biochemical modulation targeting topoisomerase I and II in human tumor xenografts in nude mice
Authors: Kim Ryungsa , Hirabayashi Naoki , Nishiyama Masahiko , Jinushi Kazuto , Toge Tetsuya , Okada Kosuke
Published: February 2007

Topoisomerase (topo) I and II are nuclear enzymes which are novel targets of cancer chemotherapy. A new camptothecin (CRT) analog, 7‐ethyl‐10‐[4‐(I‐piperidino)‐I‐piperidino]carbonyl‐oxy‐CPT (CRT‐11), is a topo‐I inhibitor with a higher activity and less toxicity than CPT. To investigate topo‐I and‐II‐targeting chemotherapy in an in vivo model, we studied the effect of sequential or co‐treatment using CPT‐11 and adriamycin (ADR) a topo‐II inhibitor, in 6 human tumor xenografts (2 esophageal, 2 gastric and 2 colon tumor lines). In sequential treatment, adriamycin was administered i.v. 24 hr after CPT‐11 treatment, and no antagonistic effect of this treatment schedule was observed. ADR cytotoxicity was potentiated significantly by CPT‐11 pretreatment in the case of 2 esophageal and 2 gastric tumor lines and I colon tumor line. On the other hand, co‐treatment abolished the sensitivity to CPT‐11 and ADR in all 6 tumor lines. Moreover, CPT‐11 did not significantly enhance the cytotoxicity of other agents tested, including mitomycin C (MMC) and cisplatin (CDDP). Flow cytometry and dot‐blot analyses showed that CPT‐11 pretreatment induced an increase in the S‐phase cell population with an increase of topo‐II mRNA expression after 24 and 48 hr, respectively, in the esophageal and colon tumor lines. These results suggest that CPT‐11 can modulate topo‐II levels to enhance the effect of topo‐II inhibitors in some human tumors, and this suggests a new clinical method of topo‐I and‐II targeting chemotherapy for human solid tumors.
Effect of TGF‐β1 on the ebv‐induced transformation of human lymphocyte cultures
Authors: Altiok Aysin , Bejarano Maria Teresa , Klein George , Klein Eva
Published: February 2007

We tested the effect of TGF‐β1 on the EBV‐induced activation and immortalization of human B lymphocytes. In lymphocyte cultures of EBV‐seropositive individuals, T cells can inhibit the EBV‐induced transformation of B cells. We found that in the presence of TGF‐β1 the transformation was more efficient, due to the inhibition of this function. TGF‐β1 inhibited the EBV‐induced proliferation of purified B‐cell cultures. Its effect was strongest when added at the beginning of the culture. Added later, the inhibition gradually decreased and was lost by the 4th day. After 10 to 14 days, in the cultures initiated with TGF‐β1, the number of cells was higher and they formed larger aggregates as compared with control cultures. Thus, TGF‐β1 modifies EBV‐induced transformation in a complex way. It inhibits the activation of B cells but does not affect those already activated. Once they acquire the immortalized state, the B cells are even stimulated by TGF‐β1.
Expression of the tumor‐associated mucin MUCI in an ovarian tumor cell line
Authors: Stern Linda , Palatsides Manuela , De Kretser Theonne , Ford Miriam
Published: February 2007

Epithelial sialomucins constitute a family of high‐molecular‐weight glycoproteins associated with epithelial cell surfaces. Aberrant expression of these molecules has been observed in certain types of human epithelial tumors. Members of the MUCI family of mucins isolated from different tissue types have been shown to differ in biochemical properties and in immunological reactivity. The polymerase chain reaction (PCR) has been used in a study of the MUCI mRNA expressed in ovarian tumor cells. The intron/exon structure of the ovarian mucin gene has been examined, the nucleotide sequence of regions of the cDNA 5' and 3' to a central highly repetitive region of the molecule determined and genomic clones for this mucin from the ovarian tumor cell line COLO316 have been isolated and analyzed. The results are compared with the nucleotide sequence data obtained by others for MUCI cDNA in breast and pancreatic cell lines. With a single nucleotide exception, the splicing pattern and nucleotide sequence obtained from MUCI mRNA in the ovarian cell line is the same as that of the mRNA found in breast and pancreatic cell lines. However, it appears that the use of alternate splice acceptors for intron I in the ovarian cell line studied here is independent of the specific sequence variation thought to determine splicing of MUCI in breast tumor cells.
Basic fibroblast growth factor induces proliferation of a rat pancreatic cancer cell line. Inhibition by somatostatin
Authors: Bensaïd M. , Tahiri‐Jouti N. , Cambillau C. , Viguerie N. , Colas B. , Vidal C. , Tauber J. P. , Estève J. P. , Susini C. , Vaysse N.
Published: February 2007

AR4–2J, a rat pancreatic acinar‐tumor cell line, was used to investigate long‐term effects of basic fibroblast growth factor (bFGF) and somatostatin on pancreatic cancer cells. We observed that bFGF stimulated cell proliferation when cells were cultured in serum‐free medium. The effect was dose‐dependent with half‐maximal and maximal effects at 25 pM and I nM bFGF, respectively. The somatostatin analog SMS 201–995 (SMS) decreased the growth‐promoting effect of bFGF. The maximal effect was observed at I nM SMS and the half‐maximal effect at 20 pM SMS. Characterization of bFGF receptor‐binding properties with [125I]bFGF revealed that AR4–2J cells exhibited 2 classes of bFGF binding site with respective KD values of 47 pM and 3 nM and binding capacities of 14 fmol and 0.9 pmol/106 cells. High‐affinity receptors correlated with bFGF stimulation of AR4–2J cell growth, suggesting that the effects of bFGF are receptor‐mediated.
Human C‐ERBB‐2 proto‐oncogene product as a target for bispecific‐antibody‐directed adoptive tumor immunotherapy
Authors: Nishimura Takashi , Nakamura Yoshihiko , Tsukamoto Hideo , Takeuchi Yasuhiro , Tokuda Yutaka , Iwasawa Megumi , Yamamoto Tadashi , Masuko Takashi , Hashimoto Yoshiyuki , Habu Sonoko
Published: February 2007

To develop an efficient strategy for the targeting of anti‐tumor effector cells, we prepared bispecific antibody (BsAb) containing anti‐CD3 and an anti‐c‐erbB‐2 proto‐oncogene product. The prepared BsAb specifically reacts with both c‐erbB‐2‐positive tumor cells and CD3+ CTL. Human CD4+ helper/killer T cells, induced from peripheral‐blood mononuclear cells by activation with immobilized anti‐CD3 monoclonal antibody (MAb) plus IL‐2, showed no significant cytotoxicity against tumor cells. However, treatment of human CD4+ helper/killer cells with the BsAb caused the induction of specific cytotoxicity against c‐erbB‐2‐positive tumor cells. CD4+ helper/killer cells also produced significant amounts of IL‐2 during co‐culture with c‐erbB‐2‐positive tumor cells in the presence of the BsAb. Moreover, by combination with the BsAb, CD4+ helper/killer cells showed a strong in vivo anti‐tumor effect against c‐erbB‐2 transfectant or human colon‐cancer cells implanted in nude mice. Our results strongly suggest that the c‐erbB‐2 proto‐oncogene product on human tumor cells may be a good target for BsAb‐directed adoptive tumor immunotherapy.
TGF‐β stimulation of endometrial and breast‐cancer cell growth
Authors: Croxtall J. D. , Jamil A. , Ayub M. , Colletta A. A. , White J. O.
Published: February 2007

Growth of the human endometrial carcinoma Ishikawa cell line was stimulated by transforming growth factor‐β1 when cultured in serum‐containing and chemically defined culture medium. This response was not unique to endometrial carcinoma cells, as the breast‐cancer cell line, T47‐D, was similarly stimulated by TGF‐β1. TGF‐β1 stimulated growth of MCF‐7 breast‐cancer cells in chemically defined medium but inhibited growth of this cell fine in serum‐containing medium. The data provide a demonstration of a positive growth response to TGF‐β1 in oestrogen‐receptor‐positive cells and do not support the hypothesis that this growth factor is simply a negative growth regulator in epithelial‐cancer cell lines.
Enhancement by bombesin of colon carcinogenesis and metastasis induced by azoxymethane in wistar rats
Authors: Iishi Hiroyasu , Tatsuta Masaharu , Baba Miyako , Yamamoto Reiko , Taniguchi Haruo
Published: February 2007

The effects of bombesin on the incidence, number and histology of colon tumors induced by azoxymethane (AOM), and on their metastases to the peritoneum and/or lymph nodes, were investigated in Wistar rats. Rats received weekly s.c. injections of AOM for 10 weeks, and s.c. injections of bombesin in depot form every other day until the end of the experiment in week 30. Administration of bombesin significantly increased the incidence of colon tumors in week 30. It had no influence on the histological features or depths of involvement of colon adenocarcinomas, but significantly increased the incidence of cancer metastasis to the peritoneum and/or lymph nodes. It also caused a significant increase in the labeling index of the colon epithelial cells. Our findings indicate that bombesin enhances the development and metastasis of colon tumors, and that this effect may be related to its effect in increasing proliferation of epithelial cells of the colon.
Basal‐cell carcinoma in temperate and tropical Australia
Authors: Czarnecki D. , Collins N. , Meehan C. , O'Brien T. , Leahy S. , Nash C.
Published: February 2007

The clinical features of patients with basal‐cell carcinoma (BCC) living in temperate and tropical Australia were studied. In the temperate zone the patients were considerably older on average, men outnumbered women and there were differences between the sexes in the site of the prevalent BCC. In the tropics the patients were significantly younger on average, the male‐to‐female ratio approached unity and there were no differences between the sexes with regard to the site of the BCC.
Tumorigenicity, mucin production and AM‐3 epitope expression in clones selected from the HT‐29 colon carcinoma cell line
Authors: Hanski C. , Stolze B. , Riecken E. G.
Published: February 2007

Two mucin‐producing cell clones (16.2 and 12.2) and a mucin‐deficient clone (15.2) were selected from the established human adenocarcinoma cell line HT‐29 by limiting dilution and Alcian blue staining. The amounts of the mucin antigen detectable on the cell surface with the monoclonal antibody (MAb) AM‐3 decreased in the order HT‐29 > 16.2 > 12.2 > 15.2 = 0. The binding avidity of AM‐3 antibody to cells as well as to mucin extracts from each cell line decreased in the same order, indicating that the epitope density on the cell‐bound mucins was highest in HT‐29 and lowest in 12.2 cells. The parental line and the mucin‐producing cell clones 16.2 and 12.2 showed no contact inhibition and grew as aggregates, while the 15.2 cells were well spread and formed a regular monolayer. The mucin‐producing cell lines injected into nude mice yielded solid tumors with different growth rates (HT‐29 > 16.2 > 12.2), while the 15.2 cell clone was not tumorigenic at all. The relative amounts of total mucin‐bound hexoses and of the mucin epitope AM‐3 decreased in the xenografts in the order HT‐29 > 16.2 > 12.2. The present system is suitable for investigating the role of mucins in growth of colon carcinoma cells and indicates that increased tumorigenicity in nude mice coincides with the increase in total mucin expression and the expression of the AM‐3 mucin epitope in tumor tissue.
Identification of high‐density lipoprotein in serum to determine anti‐cancer efficacy of doxorubicin in HeLa cells
Authors: Yung Benjamin Yat‐Ming , Bor Amy Meei‐Shuu
Published: February 2007

The cytotoxic effects of doxorubicin (DOX) and daunorubicin (DAU) on HeLa cells cultured under different serum conditions were analyzed by the “nucleophosmin translocation” assay using immunofluorescence. Bright nucleolar fluorescence was observed in untreated cells. A shift from nucleolar to nuclear fluorescence was observed with increasing doses of DOX or DAU, with longer incubation times. A lesser degree of nucleophosmin translocation from nucleoli to nucleoplasm was observed in serum‐deprived cells under the same DOX or DAU treatment. These results correlated well with those of cell‐growth‐reversibility and colony‐formation studies, showing decreased inhibitory effects of growth on cells cultured in medium without serum. Furthermore, cells cultured in medium supplemented with the lipoprotein‐free serum responded to DOX in a similar way to cells cultured without serum. High‐density lipoprotein (HDL) and low‐density lipoprotein (LDL) were then added to the lipoprotein‐free serum. Cells cultured in medium with the HDL‐supplemented, serum showed increased sensitivity to DOX. Inhibition of cell growth and colony formation was observed in such HDL‐supplemented cells upon DOX treatment (30 min). LDL, on the other hand, did not show an increase in the anti‐cancer response. These results suggested that the variation in response of cells to DOX anti‐cancer treatment under different growth conditions may be due to their varied concentrations of HDL. “Nucleophosmin translocation”, which is useful for monitoring and ensuring the efficacy of the drug during anti‐cancer treatment, provides an improved potential for successful chemotherapy.
Anti‐idiotype vaccine against Hodgkin's lymphoma: Induction of B‐ and T‐cell immunity across species barriers against cd30 antigen by murine monoclonal internal image antibodies
Authors: Pohl C. , Renner C. , Schwonzen M. , Sieber M. , Lorenz P. , Pfreundschuh M. , Diehl V.
Published: February 2007

Murine monoclonal antibody HRS‐4 (Ab,), which defines the cell‐bound and soluble CD30 antigen associated with Hodgkin's lymphoma, was used to generate monoclonal anti‐idiotypic antibodies (Ab2) in syngeneic BALB/c mice. Murine monoclonal Ab2 14G9 and Ab2 9G10 directed against HRS‐4 were shown to be anti‐idiotypic Ab2β carrying the internal image of the CD30 antigen. These antibodies bound specifically to HRS‐4 and effectively inhibited binding of HRS‐4 to a CD30 antigen preparation at concentrations as low as 50 ng/ml. KLH‐coupled Ab2β 14G9 and 9G10 induced in BALB/c mice and New Zealand white rabbits a specific polyclonal humoral response against the 120 kDa band of the CD30 antigen. Moreover, BALB/c mice immunized i.p. with KLH‐coupled 14G9 and 9G10 exhibited a statistically significant (p < 0.01) delayed‐type hyper‐sensitivity reaction against CO30 expressing Hodgkin‐derived L540‐cells. We conclude from these data that Ab2β* 14G9 and 9G10, mimicking structures of the nominal CD30 antigen, are capable of inducing a CD30‐specific T‐cell‐ and B‐cell‐mediated immune response in mice and even across species barriers in rabbits. These CD30 anti‐id antibodies may hold promise for use as vaccines against CD30‐antigen‐expressing lymphomas.
Enhancement by thyroxine of experimental carcinogenesis induced in rat colon by azoxymethane
Authors: Iishi Hiroyasu , Tatsuta Masaharu , Baba Miyako , Okuda Shigeru , Taniguchi Haruo
Published: February 2007

The effect of thyroxine (T4) on the incidence, number and histology of colon tumors induced by azoxymethane (AOM), and on the labeling index of colon mucosa were investigated in Wistar rats. Rats were given AOM by injection once a week for 10 weeks, together with T4 in depot form until the end of the experiment. Administration of T4 resulted in a significant increase in the incidence of colon tumors in week 35. However, it did not influence the histological appearance of the colon tumors or the histological types and depths of involvement of colon adenocarcinomas. Furthermore, it caused a significant increase in the labeling index of the colon during, but not after, AOM treatment. Our findings indicate that T4 enhances the development of colon tumors, which may be related to its effect in increasing proliferation of epithelial cells in the colon mucosa during administration of the carcinogen.
Differential expression of myogenic regulatory genes, MyoD1 and myogenin, in human rhabdomyosarcoma sublines
Authors: Hosoi Hajime , Sugimoto Tohru , Hayashi Yasuhide , Inaba Toshiya , Horii Yoshihiro , Morioka Hiroyuki , Fushiki Shinji , Hamazaki Minoru , Sawada Tadashi
Published: February 2007

A cell line (SCMC‐MM‐1) was established from a human abdominal tumor that was initially diagnosed as a malignant mesenchymoma by histological, immunohistochemical and clinical criteria. The cell line was composed of 2 morphologically and immunohistochemically distinct cell types, one with a small polygonal phenotype (P‐type), characterized by the immune‐staining of vimentin and the presence of a few electron‐microscopically visible organelles, and the other with a giant tubular phenotype (T‐type), characterized by the immunostaining of desmin, α‐sarcomeric actin and skeletal‐muscle myosin, and the presence of thick and thin myofilaments and Z‐line materials. The parental cell line was cloned into 2 sublines, a P‐type clone (SCMC‐MM‐1‐19P) and a T‐type clone (SCMC‐MM‐1–1T), which shared both 2q37 and 11p15 translocations, the characteristic chromosomal aberrations for rhabdomyosar‐coma, with the parental SCMC‐MM‐I cell line. Northern‐blot analyses of the myogenic regulatory genes, including MyoDI and myogenin, demonstrated the expression of MyoDI in both of these sublines. Myogenin was very weakly expressed in the SCMC‐MM‐1–19P subline, but strongly expressed in the SCMC‐MM‐1–1T subline. Chromosomal and myogenic‐regulatory‐gene analyses revealed that both of these sublines were rhabdomyo‐sarcoma cell lines. Furthermore, the regulatory‐gene analyses indicated that these 2 sublines represented 2 distinct differentiation stages of myoblasts, and that MyoD1 and myogenin could serve as the lineage marker and the differentiation marker, respectively, of human rhabdomyosarcoma.
Relationships of DNA ploidy, S‐phase fraction and hormone receptor status to tumor stage in breast cancers detected by population screening
Authors: Stål Olle , Brisfors Ann , Carstensen John , Ferraud Lilianne , Hatschek Thomas , Nordenskjöld Bo
Published: February 2007

Cellular DNA content was analyzed by flow cytometry and estrogen and progesterone receptors by an immuno‐biochemical method (EIA) in a consecutive series of 807 frozen breast‐cancer samples. Before the beginning of the study, a mammography screening program had been introduced in the region where the tumors were diagnosed. Forty percent of the tumors were judged as DNA diploid, of which 86% were ER‐positive. The proportion of ER‐positive tumors among non‐diploids was significantly lower, or 73% (p < 0.001). S‐phase fraction (SPF) was estimated in 691 cases (86%), with an overall mean of 8.4%. DNA ploidy as well as ER and PR status were independently related to SPF. Unlike the results obtained in most older series, the biological variables correlated significantly with tumor staging factors such as lymph‐node status and tumor size. Patients with nodal involvement, especially those with 4 positive nodes or more, more often had tumors which were receptor‐negative, DNA aneuploid and of high S‐phase rate. Large tumor size was significantly related to lower frequencies of receptor positivity and strongly related to DNA aneuploidy and high S‐phase fraction. Multiple linear regression analysis showed that these relationships were mainly due to the associations of SPF with the other variables. S‐phase fraction was the only independent factor predicting nodal status, while DNA ploidy in addition to SPF was associated with tumor size. In fact, DNA ploidy (p < 0.001), ER and PR status (p < 0.001, p = 0.002), nodal status (p = 0.04) and tumor size (p < 0.001) were all independently related to SPF.
Anti‐tumor and immunomodulatory activity of intraperitoneal IFN‐γ in ovarian carcinoma patients with minimal residual tumor after chemotherapy
Authors: Colombo N. , Peccatori F. , Paganin C. , Bini S. , Brandely M. , Mangioni C. , Mantovani A. , Allavena P.
Published: February 2007

Eight patients with epithelial ovarian carcinoma persisting after chemotherapy, selected for having a residual tumor no larger than I cm in diameter, were treated intra‐peritoneally (i.p.) with recombinant interferon‐γ twice weekly for 3 months. Toxicity consisted of fever and malaise in all patients and a transient rise in hepatic enzyme levels in 3 patients. The cytotoxic function of peripheral blood and peritoneal tumor‐associated lymphocytes (TAL) and macrophages (TAM), was studied using cell lines as targets. I.p. IFN‐γ augmented the cytotoxic activity of lymphocytes and mononuclear phagocytes: stimulation was more marked and more frequently observed with TAL and occasionally TAM than with blood effectors, suggesting preferential modulation at the site of tumor growth and IFN administration. Surgical laparotomy revealed that I patient had a complete response, 2 a partial response and 2 had stable disease, while 3 patients had progressive disease. In this small series of patients there was no obvious, strict correlation between immunomodulation by IFN‐γ and clinical response. These results indicate that, in contrast to its lack of activity in advanced ovarian carcinoma, IFN‐γ has definite immunomodulatory and antitumor activity in the presence of limited tumor burden.
In vitro response of human small‐cell lung‐cancer cell lines to chemotherapeutic drugs; no correlation with clinical data
Authors: Smit E. F. , de Vries E. G. E. , Timmer‐Bosscha H. , de Leij L. F. H. M. , Oosterhuis J. W. , Scheper R. J. , Weening J. J. , Postmus P. E. , Mulder N. H.
Published: February 2007

Three cell lines derived from small‐cell lung carcinoma (SCLC) tumors of patients who had no clinical response after treatment with a multi‐drug regimen were compared to 3 cell lines derived from tumors of patients who, upon treatment, showed a complete clinical response. These 2 groups of cell lines were considered to represent the in vitro counterparts of the 2 extremes of the clinical spectrum of sensitivity for chemotherapeutic drugs in small‐cell lung cancer. To assess whether the in vivo (in)sensitivity of a tumor to a certain drug regimen is retained in vitro, the cell lines were tested for drug sensitivity using the microtiter‐well tetrazolium assay and the results were compared with the in vivo data. No correlation was found. Since in vitro models using cell lines are based on the assumption that a cell line reflects the properties of the tumor from which it is derived, several additional parameters such as MAb staining against different SCLC‐associated antigens and DNA content were analyzed in the biopsies and the cell lines. The results showed that selection of discrete tumor‐cell populations in vitro occurs. Results of in vitro chemosensitivity testing for individual SCLC patients should be interpreted with caution.