bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Macrophages Promote the Motility and Invasiveness of Osteopontin-Knockdown Tumor Cells
Authors: Cheng Jiasen , Huo De-Hua , Kuang Dong-Ming , Yang Jine , Zheng Limin , Zhuang Shi-Mei
Published: June 2007

Increasing evidence indicates that macrophages in tumor stroma can significantly modify the malignant phenotypes of tumors. Osteopontin (OPN) is frequently overexpressed in cancers with high metastatic capacity and, thus, has been considered as a potential therapeutic target. To find out whether macrophages can affect the outcome of OPN-knockdown tumor cells, we used RNA interference (RNAi) to stably silence the OPN expression in the highly invasive human hepatoma cell line SK-Hep-1. Silencing of OPN markedly decreased the motility and invasiveness of the SK-Hep-1 cells. Further studies using this cell model revealed that coculture with human macrophages or macrophage-conditioned medium largely restored the migration and invasion potential of OPN-knockdown tumor cells. Moreover, such macrophage-promoted motility can be effectively blocked either by the addition of OPN-neutralizing antibody to the cocultured medium or by silencing OPN expression in macrophages. These results indicate that macrophage-derived OPN can compensate for the decrease of OPN and thereby restore the metastatic potential of OPN-knockdown tumor cells. Further characterization of the underlying mechanisms disclosed that macrophage-derived OPN exerted its function independently of the actin cytoskeleton rearrangement or the activation of matrix metalloproteinase and Rho families. Our results suggest that there are fine-tuned complex interactions between cancer cells and stroma cells, which may modify the outcome of cancer therapy, and therefore should be considered for the rational design of anticancer strategy. [Cancer Res 2007;67(11):5141–7]
Eradication of Therapy-Resistant Human Prostate Tumors Using a Cancer Terminator Virus
Authors: Sarkar Devanand , Lebedeva Irina V. , Su Zao-zhong , Park Eun-Sook , Chatman Lejuan , Vozhilla Nicollaq , Dent Paul , Curiel David T. , Fisher Paul B.
Published: June 2007

Terminal prostate cancer is refractory to conventional anticancer treatments because of frequent overexpression of antiapoptotic proteins Bcl-2 and/or Bcl-xL. Adenovirus-mediated delivery of melanoma differentiation associated gene-7/interleukin-24 (mda-7/IL-24), a secreted cytokine having cancer-selective apoptosis-inducing properties, profoundly inhibits prostate cancer cell growth. However, forced overexpression of Bcl-2 or Bcl-xL renders prostate cancer cells resistant to Ad.mda-7. We constructed a conditionally replication-competent adenovirus in which expression of the adenoviral E1A gene, necessary for replication, is driven by the cancer-specific promoter of progression elevated gene-3 (PEG-3) and which simultaneously expresses mda-7/IL-24 in the E3 region of the adenovirus (Ad.PEG-E1A-mda-7), a cancer terminator virus (CTV). This CTV generates large quantities of MDA-7/IL-24 as a function of adenovirus replication uniquely in cancer cells. Infection of Ad.PEG-E1A-mda-7 (CTV) in normal prostate epithelial cells and parental and Bcl-2– or Bcl-xL–overexpressing prostate cancer cells confirmed cancer cell–selective adenoviral replication, mda-7/IL-24 expression, growth inhibition, and apoptosis induction. Injecting Ad.PEG-E1A-mda-7 (CTV) into xenografts derived from DU-145-Bcl-xL cells in athymic nude mice completely eradicated not only primary tumors but also distant tumors (established in the opposite flank), thereby implementing a cure. These provocative findings advocate potential therapeutic applications of this novel virus for advanced prostate cancer patients with metastatic disease. [Cancer Res 2007;67(11):5434–42]
The Soluble Wnt Receptor Frizzled8CRD-hFc Inhibits the Growth of Teratocarcinomas In vivo
Authors: DeAlmeida Venita I. , Miao Li , Ernst James A. , Koeppen Hartmut , Polakis Paul , Rubinfeld Bonnee
Published: June 2007

Wnt signaling is important for normal cell proliferation and differentiation, and mutations in pathway components are associated with human cancers. Recent studies suggest that altered wnt ligand/receptor interactions might also contribute to human tumorigenesis. Therefore, agents that antagonize wnt signaling at the extracellular level would be attractive therapeutics for these cancers. We have generated a soluble wnt receptor comprising the Frizzled8 cysteine-rich domain (CRD) fused to the human Fc domain (F8CRDhFc) that exhibits favorable pharmacologic properties in vivo. Potent antitumor efficacy was shown using the mouse mammary tumor virus-Wnt1 tumor model under dosing conditions that did not produce detectable toxicity in regenerating tissue compartments. In vitro, F8CRDhFc inhibited autocrine wnt signaling in the teratoma cell lines PA-1, NTera-2, Tera-2, and NCCIT. In vivo, systemic administration of F8CRDhFc significantly retarded the growth of tumor xenografts derived from two of these cell lines, PA-1 and NTera-2. Pharmacodynamic markers of wnt signaling, identified by gene expression analysis of cultured teratoma cells, were also modulated in the tumor xenografts following treatment with F8CRDhFc. Additionally, these markers could be used as indicators of treatment efficacy and might also be useful in identifying patients that would benefit from the therapeutic agent. This is the first report showing the efficacy of a soluble wnt receptor as an antitumor agent and suggests that further development of wnt antagonists will have utility in treating human cancer. [Cancer Res 2007;67(11):5371–9]
HS1-Associated Protein X-1 Regulates Carcinoma Cell Migration and Invasion via Clathrin-Mediated Endocytosis of Integrin αvβ6
Authors: Ramsay Alan G. , Keppler Melanie D. , Jazayeri Mona , Thomas Gareth J. , Parsons Maddy , Violette Shelia , Weinreb Paul , Hart Ian R. , Marshall John F.
Published: June 2007

Enhanced expression levels of integrin αvβ6 have been linked to more aggressive invasive carcinoma cell behavior and poorer clinical prognosis. However, how αvβ6 determines invasion and the dynamics of integrin αvβ6 regulation in tumor cells are poorly understood. We have identified the 35-kDa HS1-associated protein X-1 (HAX-1) protein as a novel binding partner of the β6 cytoplasmic tail using a yeast two-hybrid screen. We show that αvβ6-dependent migration is blocked following small interfering RNA (siRNA)–mediated depletion of HAX-1 in oral squamous cell carcinoma cell lines. Using both siRNA and membrane-permeable peptides, we show that αvβ6-dependent migration and invasion require HAX-1 to bind directly to β6 and thereby regulate clathrin-mediated endocytosis of αvβ6 integrins. Progression of oral cancer is associated with enhanced expression of αvβ6 and HAX-1 proteins in patient tissue. This report establishes that integrin endocytosis is required for αvβ6-dependent carcinoma cell motility and invasion and suggests that this process is an important mechanism in cancer progression. [Cancer Res 2007;67(11):5275–84]
The Peroxisome Proliferator WY-14,643 Promotes Hepatocarcinogenesis Caused by Endogenously Generated Oxidative DNA Base Modifications in Repair-Deficient Csbm/m/Ogg1−/− Mice
Authors: Trapp Christian , Schwarz Michael , Epe Bernd
Published: June 2007

Basal levels of endogenously generated oxidative DNA modifications such as 7,8-dihydro-8-oxoguanine (8-oxoG) are present in apparently all mammalian cells, but their relevance for the generation of spontaneous cancers remains to be established. Both the 8-oxoG levels and the resulting spontaneous mutations are increased in the livers of Csbm/m/Ogg1−/− mice, which are deficient in the repair of 8-oxoG. In order to determine the consequences of these additional oxidative DNA modifications and mutations and thus assess the tumor initiating potency of this type of endogenous DNA damage, we treated Csbm/m/Ogg1−/− mice and repair-proficient controls with the peroxisome proliferator WY-14,643 (0.025% ad libitum), a potent inducer of liver cell proliferation. The treatment did not generate any additional oxidative DNA damage; the elevated levels of 8-oxoG in the Csbm/m/Ogg1−/− mice even decreased. Also, the spontaneous mutation frequencies observed in the lacI gene of BigBlue Csbm/m/Ogg1−/− mice, which were ∼3-fold higher than in the repair-proficient mice, declined by 39% under the treatment, whereas the frequencies in the livers of the repair-proficient animals remained unchanged. Preneoplastic lesions (staining positive or negative for glucose-6-phoshatase) developed in the livers of both wild-type and Csbm/m/Ogg1−/− mice after 30 weeks. Both the numbers and the total volumes of the lesions were ∼6-fold higher in the repair-deficient mice than in the wild-type mice. The results indicate that spontaneous mutations generated from endogenous oxidative DNA base damage efficiently translate into increased tumorigenesis when cell proliferation is stimulated. [Cancer Res 2007;67(11):5156–61]
Myc Goes Global: New Tricks for an Old Oncogene
Authors: Knoepfler Paul S.
Published: June 2007

Myc, a transcription factor commonly deregulated in tumorigenesis, is thought to mediate its diverse cellular effects by altering the expression of specific target genes. However, it has been difficult to gain a precise understanding of how Myc drives cancer because Myc acts rather weakly at many of its target loci, and it has been reported to regulate as many as 10% to 15% of all cellular genes. A new perspective on this issue has been provided by a recent study that revealed Myc can regulate chromatin structure in a global fashion. These findings suggest actions for Myc that extend beyond the traditional concept of a targeted gene regulator. [Cancer Res 2007;67(11):5061–3]
Regression of Early and Intermediate Stages of Colon Cancer by Targeting Multiple Members of the EGFR Family with EGFR-Related Protein
Authors: Schmelz Eva M. , Xu Hu , Sengupta Radha , Du Jianhua , Banerjee Sanjeev , Sarkar Fazlul H. , Rishi Arun K. , Majumdar Adhip P.N.
Published: June 2007

A role of the epidermal growth factor receptor (EGFR) family has been suggested in colon cancer etiology, progression, and/or severity. Our recently identified pan-erbB inhibitor EGFR-related protein (ERRP) targets EGFRs by attenuating their activation and subsequent signaling leading to cellular growth inhibition. In the present study, we evaluated the therapeutic effectiveness of ERRP on early and intermediate stages of colon cancer by examining regression of chemically induced aberrant crypt foci (ACF) in the colon of CF1 mice and intestinal adenomas in APCMin+/− (Min) mice. After formation of ACF or adenomas, the mice were injected (i.p.) with ERRP (50 μg/mouse) for 10 consecutive days. This treatment significantly reduced the number of ACF from 25.0 ± 3.0 (controls) to 14.9 ± 1.6 (ERRP-treated; P = 0.011) and also reduced their size (P < 0.01). In Min mice, ERRP caused the regression of adenomas throughout the small intestine (P < 0.05) and reduced their size (P < 0.001). This could partly be attributed to inhibition of proliferation and stimulation of apoptosis in the intestinal mucosa and was associated with decreased activation of several EGFR family members, suppression of downstream effector nuclear factor κB and down-regulation of cyclooxygenase-2. ERRP-induced attenuation of EGFR activation could be due to increased sequestration of the ligand(s) by ERRP, rendering them unavailable for binding to and activation of the receptor. In conclusion, our data show that ERRP is effective in regressing both early and intermediate intestinal lesions and could be an effective therapeutic agent for colon cancer. [Cancer Res 2007;67(11):5389–96]
Sequential Treatment by Ionizing Radiation and Sodium Arsenite Dramatically Accelerates TRAIL-Mediated Apoptosis of Human Melanoma Cells
Authors: Ivanov Vladimir N. , Zhou Hongning , Hei Tom K.
Published: June 2007

Melanoma is the most lethal form of skin cancer. There is a lack of effective treatments for individuals with advanced disease. Many melanomas exhibit high levels of radioresistance. The direct consequence of γ-irradiation for most melanoma cells is growth arrest at the G2-M phase of cell cycle. However, radiation-induced signaling pathways may affect numerous additional targets in cancer cells. We show in the present study that γ-irradiation, as well as α-particle exposure, dramatically increases the susceptibility of melanoma cells to recombinant tumor necrosis factor–related apoptosis-inducing ligand (TRAIL)-mediated apoptosis via up-regulation of surface TRAIL-receptor 1/receptor 2 (DR4/DR5) levels and to Fas ligand–mediated apoptosis via up-regulation of surface Fas levels. Additionally, increased dynamin-2 expression after irradiation is critically important in the translocation of death receptor to the cell surface. Moreover, sodium arsenite treatment may up-regulate expression of endogenous TRAIL and induces its translocation to cell surface and further down-regulates cFLIP levels in melanoma cells. We have evaluated the effects of sequential γ-irradiation and arsenite treatment of melanoma cells for the induction of death signaling. Such treatment results in an efficient TRAIL-mediated apoptosis via a paracrine mechanism. These data highlight the efficacy of combined modality treatment involving radiation and arsenite in clinical management of this often fatal form of skin cancer. [Cancer Res 2007;67(11):5397–407]
Tumor-Associated Macrophages Press the Angiogenic Switch in Breast Cancer
Authors: Lin Elaine Y. , Pollard Jeffrey W.
Published: June 2007

The development of a supportive vasculature is essential for tumor progression. In a mouse model of breast cancer, we found that tumor-associated macrophages that are recruited to the tumor just before malignant conversion are essential for the angiogenic switch. These findings establish a causal linkage to explain well-documented clinical correlations between macrophages, microvessel density, and poor prognosis in breast tumors. [Cancer Res 2007;67(11):5064–3]
Differential Regulation of Estrogen Receptor α Turnover and Transactivation by Mdm2 and Stress-Inducing Agents
Authors: Duong Vanessa , Boulle Nathalie , Daujat Sylvain , Chauvet Jérôme , Bonnet Sandrine , Neel Henry , Cavaillès Vincent
Published: June 2007

In mammalian cells, the level of estrogen receptor α (ERα) is rapidly decreased upon estrogen treatment, and this regulation involves proteasome degradation. Using different approaches, we showed that the Mdm2 oncogenic ubiquitin-ligase directly interacts with ERα in a ternary complex with p53 and is involved in the regulation of ERα turnover (both in the absence or presence of estrogens). Several lines of evidence indicated that this effect of Mdm2 required its ubiquitin-ligase activity and involved the ubiquitin/proteasome pathway. Moreover, in MCF-7 human breast cancer cells, various p53-inducing agents (such as UV irradiation) or treatment with RITA (which inhibits the interaction of p53 with Mdm2) stabilized ERα and abolished its 17β-estradiol–dependent turnover. Interestingly, our data indicated that ligand-dependent receptor turnover was not required for efficient transactivation. Altogether, our results indicate that the Mdm2 oncoprotein and stress-inducing agents complexly and differentially regulate ERα stability and transcriptional activity in human cancer cells. [Cancer Res 2007;67(11):5513–21]
Phase I Trial of BAY 50-4798, an Interleukin-2–Specific Agonist in Advanced Melanoma and Renal Cancer
Authors: Margolin Kim , Atkins Michael B. , Dutcher Janice P. , Ernstoff Marc S. , Smith John W. , Clark Joseph I. , Baar Joseph , Sosman Jeffrey , Weber Jeffrey , Lathia Chetan , Brunetti Janice , Cihon Frank , Schwartz Brian
Published: June 2007

Purpose: BAY 50-4798 is an analogue of interleukin-2 that selectively activates T cells over natural killer cells. This phase I study was designed to determine the maximum tolerated dose (MTD) and safety of BAY 50-4798, screen for tumor response, and assess pharmacokinetics.

Experimental Design: Forty-five patients with metastatic melanoma or renal cancer were enrolled, 31 on escalating doses to determine the MTD, with 20 renal cell carcinoma patients treated at MTD to detect antitumor activity. BAY 50-4798 was delivered i.v. every 8 h, days 1 to 5 and 15 to 19, and could be repeated after 9 weeks if tumor was stable or responding.

Results: The MTD was defined by and reported in terms of doses received. The doses tested ranged from 1.3 to 26.1 μg/kg, and the MTD was defined as 10.4 μg/kg based on toxicities similar to those of aldesleukin. Two patients achieved partial responses, one with melanoma and one with renal cell carcinoma. Among all 45 patients, 53% and 9% experienced a grade 3 and 4 toxicity, respectively. Among the patients treated at the MTD of 10.4 μg/kg, 71% and 10% experienced a grade 3 and 4 toxicity, respectively. Pharmacokinetics showed dose-dependent peak concentrations (Cmax) and area under the curve with a half-life of ∼2 h and no evidence of accumulation. Lymphocyte subset analysis confirmed the preferential expansion of T-cell subsets over natural killer cells.

Conclusions: The antitumor activity of BAY 50-4798 in malignancies that respond to high-dose interleukin-2 was low. BAY 50-4798 might provide advantages over aldesleukin in antigen-specific immunotherapies.
Interaction between Cancer Cells and Stromal Fibroblasts Is Required for Activation of the uPAR-uPA-MMP-2 Cascade in Pancreatic Cancer Metastasis
Authors: He Yu , Liu Xiang-de , Chen Zhi-yu , Zhu Jin , Xiong Yan , Li Kun , Dong Jia-hong , Li Xiaowu
Published: June 2007

Purpose: Interaction between tumor cells and surrounding stromal fibroblast (SF) plays a critical role in tumor growth and invasion. The aim of the study is to determine the role of SF in regulating the invasive behaviors of pancreatic cancer by evaluating the mode of SF activating the urokinase plasminogen activator (uPA)-plasmin-matrix metalloproteinase (MMP)-2 cascade.

Experimental Design: The expression patterns of uPA, MMP-2, and uPA receptor (uPAR) in human metastatic pancreatic cancer were analyzed by immunohistochemistry and the roles of SF in activation of the uPA-plasmin-MMP-2 cascade were evaluated by coculturing pancreatic cancer cell lines with SF.

Results: uPA expression and fibroblastic uPAR expression were correlated with liver metastasis of human pancreatic cancer. MMP-2 rather than MMP-9 was activated in the metastatic pancreatic cancer. In the in vitro culture system, the coculture of peritumor fibroblasts with metastatic pancreatic cancer BxPc3 cells resulted in activation of MMP-2 and up-regulation of uPAR expression. In this coculture system, the uPA-plasminogen cascade was involved in MMP-2 activation. This activation required a direct interaction between SF and cancer cells. In the coculture system, intergrin α6β1 expression was increased in BxPc3 cells, and blocking the function of integrin α6β1 decreased the activation of uPA and MMP-2. This suggests that interaction between integrins of cancer cells and the uPARs of the SF might be involved in the activation of the uPAR-uPA-MMP-2 cascade.

Conclusion: Our results suggest that SF plays a role in promoting pancreatic cancer metastasis via activation of the uPA-plasminogen-MMP-2 cascade.
In vivo Therapeutic Synergism of Anti–Epidermal Growth Factor Receptor and Anti-HER2 Monoclonal Antibodies against Pancreatic Carcinomas
Authors: Larbouret Christel , Robert Bruno , Navarro-Teulon Isabelle , Thèzenas Simon , Ladjemi Maha-Zohra , Morisseau Sébastien , Campigna Emmanuelle , Bibeau Frédéric , Mach Jean-Pierre , Pèlegrin André , Azria David
Published: June 2007

Purpose: Pancreatic carcinoma is highly resistant to therapy. Epidermal growth factor receptor (EGFR) and HER2 have been reported to be both dysregulated in this cancer. To evaluate the in vivo effect of binding both EGFR and HER2 with two therapeutic humanized monoclonal antibodies (mAb), we treated human pancreatic carcinoma xenografts, expressing high EGFR and low HER2 levels.

Experimental Design: Nude mice, bearing xenografts of BxPC-3 or MiaPaCa-2 human pancreatic carcinoma cell lines, were injected twice weekly for 4 weeks with different doses of anti-EGFR (matuzumab) and anti-HER2 (trastuzumab) mAbs either alone or in combination. The effect of the two mAbs, on HER receptor phosphorylation, was also studied in vitro by Western blot analysis.

Results: The combined mAb treatment significantly inhibited tumor progression of the BxPC-3 xenografts compared with single mAb injection (P = 0.006) or no treatment (P = 0.0004) and specifically induced some complete remissions. The two mAbs had more antitumor effect than 4-fold greater doses of each mAb. The significant synergistic effect of the two mAbs was confirmed on the MiaPaCa-2 xenograft and on another type of carcinoma, SK-OV-3 ovarian carcinoma xenografts. In vitro, the cooperative effect of the two mAbs was associated with a decrease in EGFR and HER2 receptor phosphorylation.

Conclusions: Anti-HER2 mAb has a synergistic therapeutic effect when combined with an anti-EGFR mAb on pancreatic carcinomas with low HER2 expression. These observations may open the way to the use of these two mAbs in a large panel of carcinomas expressing different levels of the two HER receptors.
Effect of Positive Bone Marrow EBV In situ Hybridization in Staging and Survival of Localized Extranodal Natural Killer/T-Cell Lymphoma, Nasal-Type
Authors: Lee Jeeyun , Suh Cheolwon , Huh Jooryung , Jun Hyun Jung , Kim Kihyun , Jung Chulwon , Park Keunchil , Park Yeon Hee , Ko Young Hyeh , Kim Won Seog
Published: June 2007

Purpose: The aim of the study was to determine the effect of EBV-encoded RNA-1 in situ hybridization (EBER-1 ISH) in bone marrow specimens on survival outcome in patients with clinical stage I/II natural killer/T-cell lymphoma.

Experimental Design: We systematically did EBER-1 ISH on 182 archival bone marrow tissues from 91 patients who were diagnosed of stage I/II natural killer/T-cell lymphoma and analyzed the correlation between bone marrow EBER-1 ISH status and survival. We defined minimal bone marrow involvement and definite bone marrow involvement to distinguish the subgroups who revealed EBV-positive cells from normal marrow by EBER-1 ISH from those who showed typical neoplastic cells in bone marrow biopsies.

Results: In total, 17 of the 91 (18.7%) patients showed positivity for EBER-1 ISH at least in one of the bilateral bone marrow biopsies with 14 minimal bone marrow involvements and 3 definite bone marrow involvements. Patients with positive bone marrow EBER-1 ISH showed significantly poorer overall survival than those who were negative for bone marrow EBER-1 ISH (median survival, 16.1 months versus not reached; P = 0.045).

Conclusion: Considering a high proportion of stage I/II patients (15.4%) with minimal in bone marrow specimens, bone marrow EBER-1 ISH should be routinely done in all patients with localized disease for more accurate staging.
Phase I and Pharmacokinetic Study of AI-850, A Novel Microparticle Hydrophobic Drug Delivery System for Paclitaxel
Authors: Mita Alain C. , Olszanski Anthony J. , Walovitch Richard C. , Perez Raymond P. , MacKay Kathleen , Tuck David P. , Simmons Cecilia , Hammond Susan , Mita Monica M. , Beeram Muralidhar , Stone Anne J. , Rowinsky Eric K. , Lewis Lionel D.
Published: June 2007

Purpose: AI-850, paclitaxel in a novel polyoxyethylated castor oil-free hydrophobic microparticle delivery system, is being developed based on its favorable preclinical safety and antitumor activity profiles. The objectives of the study were to assess the feasibility and safety of administering AI-850 as a <30-min i.v. infusion without premedication every 3 weeks, determine the maximum tolerated dose and the phase II recommended dose of AI-850, study the pharmacokinetics of paclitaxel in this new formulation, and seek evidence of anticancer activity.

Experimental Design: This was an open-label phase I dose escalation study of AI-850 in patients with advanced solid malignancies. AI-850 doses were escalated according to a modified Fibonacci scheme. Clinical and laboratory toxicity was monitored, and paclitaxel plasma concentrations were measured by liquid chromatography-tandem mass spectrometry.

Results: Twenty-two patients received 56 courses of AI-850 at five dose cohorts ranging from 36 to 250 mg/m2. Grade 4 neutropenia, either exceeding 5 days or complicated by fever, was dose limiting in two of six patients at 250 mg/m2 AI-850. Three patients experienced grade 2 to 4 infusion-related adverse reactions. Toxicities, including fatigue, alopecia, nausea and vomiting, neuropathy, anorexia, and myalgia, were mild to moderate, reversible, and not dose related. Pharmacokinetics of free and total paclitaxel showed biexponential plasma decay and dose proportionality for maximum plasma paclitaxel concentration and area under the concentration versus time curve. Antitumor activity was documented in two patients with endometrial and tongue carcinomas.

Conclusions: The administration of AI-850 as a brief infusion once every 3 weeks was feasible at doses up to 205 mg/m2. The potential of AI-850 as an alternative to other approved paclitaxel formulations requires further clinical evaluation.
Association between the Estrogen Receptor α A908G Mutation and Outcomes in Invasive Breast Cancer
Authors: Herynk Matthew H. , Parra Irma , Cui Yukun , Beyer Amanda , Wu Meng-Fen , Hilsenbeck Susan G. , Fuqua Suzanne A.W.
Published: June 2007

Purpose: Estrogen receptor α (ERα) predicts the natural history of breast cancer without intervening therapy. Here, we have optimized the detection of a somatic mutation, an A908G transition of ERα, and examined its association with clinical and biological features of invasive breast cancer.

Experimental Design: We compared two methods of sequencing to detect the A908G ERα mutation. We then used primer extension sequencing with genomic DNA isolated from invasive breast tumors to determine whether the mutation was associated with clinical outcome in 267 axillary node–negative and axillary node–positive breast tumors. The presence of the mutation and clinical variables were analyzed for association with recurrence-free survival and overall survival by Cox proportional hazards regression models.

Results: We determined that dye-labeled terminator sequencing was not adequate for detection of the A908G ERα mutation. The mutation was detected at a high frequency (50%) in invasive breast tumors using primer extension sequencing, and was found to be associated with clinical measures of poor outcome, including larger tumor size and axillary lymph node positivity. Although the mutation was associated with recurrence-free survival in univariate analysis, it was not an independent predictor of outcomes in multivariate analysis.

Conclusions: Consistent with our previous finding of this somatic ERα mutation in breast ductal hyperplasias, we now present evidence that the A908G mutation is present in invasive breast tumors using an optimized sequencing method. We find that the mutation is significantly associated with aggressive biological tumor features, and with an unfavorable prognosis, but was not an independent prognostic marker in untreated patients.
Cross-Linking of Signal Transducer and Activator of Transcription 3—A Molecular Marker for the Photodynamic Reaction in Cells and Tumors
Authors: Henderson Barbara W. , Daroqui Cecilia , Tracy Erin , Vaughan Lurine A. , Loewen Gregory M. , Cooper Michele T. , Baumann Heinz
Published: June 2007

Purpose: Photodynamic therapy (PDT) depends on the delivery of a photosensitizer to the target tissue that, under light exposure, produces singlet oxygen and other reactive oxygen species, which in turn cause the death of the treated cell. This study establishes a quantitative marker for the photoreaction that will predict the outcome of PDT.

Experimental Design: Cells in tissue culture, murine s.c. tumors, and endobronchial carcinomas in patients were treated with PDT, and the noncleavable cross-linking of the latent signal transducer and activator of transcription 3 (STAT3) was determined.

Results: Murine and human cancer cell lines reacted to PDT by an immediate covalent cross-linking of STAT3 to homodimeric and other complexes. The magnitude of this effect was strictly a function of the PDT reaction that is determined by the photosensitizer concentration and light dose. The cross-link reaction of STAT3 was proportional to the subsequent cytotoxic outcome of PDT. An equivalent photoreaction as detected in vitro occurred in tumors treated in situ with PDT. The light dose-dependent STAT3 cross-linking indicated the relative effectiveness of PDT as a function of the distance of the tissue to the treating laser light source. Absence of cross-links correlated with treatment failure.

Conclusions: The data suggest that the relative amount of cross-linked STAT3 predicts the probability for beneficial outcome, whereas absence of cross-links predicts treatment failure. Determination of STAT3 cross-links after PDT might be clinically useful for early assessment of PDT response.
Cytokeratin-18 Is a Useful Serum Biomarker for Early Determination of Response of Breast Carcinomas to Chemotherapy
Authors: Olofsson Maria Hägg , Ueno Takayuki , Pan Yang , Xu Ren , Cai Feng , van der Kuip Heiko , Muerdter Thomas E. , Sonnenberg Maike , Aulitzky Walter E. , Schwarz Stephan , Andersson Elina , Shoshan Maria C. , Havelka Aleksandra Mandic , Toi Masakazu , Linder Stig
Published: June 2007

Purpose: With a widening arsenal of cancer therapies available, it is important to develop therapy-specific predictive markers and methods to rapidly assess treatment efficacy. We here evaluated the use of cytokeratin-18 (CK18) as a serum biomarker for monitoring chemotherapy-induced cell death in breast cancer.

Experimental Design: Different molecular forms of CK18 (caspase cleaved and total) were assessed by specific ELISA assays. Drug-induced release of CK18 was examined from breast carcinoma cells and tissue. CK18 protein composition was examined in serum. CK18 levels were determined in serum from 61 breast cancer patients during docetaxel or cyclophosphamide/epirubicin/5-fluorouracil (CEF) therapy.

Results: Caspase-cleaved CK18 molecules were released from monolayer cultures and tumor organ cultures to the extracellular compartment. CK18 was present in complexes with other cytokeratins in serum. Such CK18 protein complexes are remarkably stable, leading to favorable performance of CK18 biomarker assays for clinical investigations. Docetaxel induced increased levels of caspase-cleaved CK18 in serum from breast cancer patients, indicating apoptosis. CEF therapy led to increases predominantly in uncleaved CK18, indicating induction of necrotic cell death in many tumors. The increase in total CK18 at 24 h of the first treatment cycle correlated to the clinical response to CEF therapy (P < 0.0001).

Conclusions: Induction of necrotic cell death may explain the clinical efficacy of anthracycline-based therapy for breast carcinomas with defective apoptosis pathways. We suggest that CK18 biomarkers are useful for early prediction of the response to CEF therapy in breast cancer and may be useful biomarkers for clinical trials.
Mifepristone Inhibits Ovarian Cancer Cell GrowthIn vitroandIn vivo
Authors: Goyeneche Alicia A. , Carón Rubén W. , Telleria Carlos M.
Published: June 2007

Purpose: These studies were designed to determine whether the synthetic steroid mifepristone inhibits ovarian cancer growth in vitro and in vivo and the molecular mechanisms involved.

Experimental Design: The effect of mifepristone on ovarian cancer cell growth in vitro was studied in ovarian cancer cell lines of different genetic backgrounds (SK-OV-3, Caov-3, OV2008, and IGROV-1). In addition, the growth inhibition capacity of mifepristone on ovarian carcinoma xenografts was tested in nude mice.

Results: Mifepristone inhibited ovarian cancer cell proliferation in a dose- and time-dependent manner. The cytostatic effect of mifepristone was confirmed in a clonogenic survival assay and was not linked to loss of viability. Mifepristone blocked DNA synthesis, arrested the cell cycle at the G1-S transition, up-regulated cyclin-dependent kinase (cdk) inhibitors p21cip1and p27kip1, down-regulated transcription factor E2F1, decreased expression of the E2F1-regulated genes cdk1 (cdc2) and cyclin A, and modestly decreased cdk2 and cyclin E levels. The abrupt arrest in cell growth induced by mifepristone correlated with reduced cdk2 activity, increased association of cdk2 with p21cip1 and p27kip1, increased nuclear localization of the cdk inhibitors, and reduced nuclear abundance of cdk2 and cyclin E. In vivo, mifepristone significantly delayed the growth of ovarian carcinoma xenografts in a dose-dependent manner and without apparent toxic effects for the animals.

Conclusions: These preclinical studies show that mifepristone is effective as a single agent in vitro and in vivo, inhibiting the growth of human epithelial ovarian cancer cells. Mifepristone markedly reduces cdk2 activity likely due to increased association of cdk2 with the cdk inhibitors p21cip1 and p27kip1 and reduced nuclear cdk2/cyclin E complex availability. Acting as a cytostatic agent, mifepristone promises to be of translational significance in ovarian cancer therapeutics.
Identification ofNTN4, TRA1, andSTC2as Prognostic Markers in Breast Cancer in a Screen for Signal Sequence Encoding Proteins
Authors: Esseghir Selma , Kennedy Alan , Seedhar Pooja , Nerurkar Ashutosh , Poulsom Richard , Reis-Filho Jorge S. , Isacke Clare M.
Published: June 2007

Purpose: In a previous screen using a signal-trap library, we identified a number of secreted proteins up-regulated in primary tumor cells isolated from invasive breast cancers. The purpose of this study was to assess the expression of these genes in human invasive breast tumors and to determine the significance of their expression for prognosis in breast cancer.

Experimental Design: A tissue microarray containing 245 invasive breast tumors from women treated with curative surgery followed by anthracycline-based chemotherapy and hormone therapy for the estrogen receptor (ER)–positive tumors was screened by in situ hybridization with probes against thrombospondin 3 (TSP3), insulin-like growth factor binding protein 7 (IGFBP7), tumor rejection antigen 1 (TRA1), stanniocalcin 2 (STC2), and netrin 4 (NTN4). Correlations between categorical variables were done using the χ2 test and Fisher's exact test. Cumulative survival probabilities were calculated using the Kaplan-Meier method and multivariate survival analysis was done with Cox hazard model. A series of breast cancers were also stained with NTN4 antibodies.

Results: All five genes examined were expressed in invasive breast tumor cells. NTN4 protein expression was also confirmed by immunohistochemistry. Together, these data validate the design and screening of the signal-trap library. Univariate survival analysis revealed that expressions of TRA1, STC2, and NTN4 are correlated with longer disease-free survival and that TRA1 and NTN4 are associated with longer overall survival. Multivariate analysis showed that NTN4 is an independent prognostic factor of overall survival.

Conclusions: This article describes the identification of three secreted proteins, NTN4, TRA1, and STC2, as potential novel prognostic markers in breast cancer.
Suppression of lung cancer tumor growth in a nude mouse model by the Ras inhibitor salirasib (farnesylthiosalicylic acid)
Authors: Zundelevich Adi , Elad-Sfadia Galit , Haklai Ronit , Kloog Yoel
Published: June 2007

Aberrant Ras pathway functions contribute to the malignant phenotype of lung cancers. Inhibitors of Ras might therefore be considered as potential drugs for lung cancer therapy. Here, we show that the Ras inhibitor farnesylthiosalicylic acid (salirasib) inhibits proliferation of human lung cancer cells harboring a mutated K-ras gene (A549, H23, or HTB54) or overexpressing a growth factor receptor (H1299 or HTB58) and enhances the cytotoxic effect of the chemotherapeutic drug gemcitabine. Salirasib inhibited active K-Ras in A549 cells, reversed their transformed morphology, and inhibited their anchorage-independent growth in vitro. Tumor growth in A549 and HTB58 cell nude mouse models was inhibited by i.p. administration of salirasib. P.o. formulated salirasib also inhibited A549 cell tumor growth. Our results suggest that p.o. salirasib may be considered as a potential treatment for lung cancer therapy. [Mol Cancer Ther 2007;6(6):1765–1773]
Oral Health among 35- to 44-Year-Old Iranians
Authors: Hessari Hossein , Vehkalahti Miira M. , Eghbal Mohammad J. , Murtomaa Heikki T.
Published: June 2007

Objective: To investigate the oral health status of 35- to 44-year-old Iranians in relation to gender, place of residence and education. Subjects and Methods: In 2002, data (n = 8,301) were collected as part of a national survey using WHO criteria for sampling and clinical diagnoses across 28 provinces by 33 calibrated examiners who performed examinations under a dental light with a WHO probe. The study sample was 8,301; male: 3,625 and female: 4,676; urban: 4,854 and rural: 3,447. Oral health status was assessed by number of decayed (DT), filled (FT) and missing (MT) teeth and by DMFT and need for periodontal treatment [community periodontal index for treatment needs (CPITN)] indices. Gender, age, place of residence and level of education served as socio-demographic information. Estimates were adjusted for the 35- to 44-year-old provincial population. ANOVA, χ2 test and logistic regression analysis with odds ratios (OR) and their 95% confidence intervals (CI) were the methods of statistical evaluation used. Results: The mean number of teeth was 21.5 ± 6.2, with DMFT: 11.0 ± 6.4, DT: 2.6 ± 2.7, and FT: 1.8 ± 3.2. DT existed in 6,080 (73%) of the subjects and FT in 3,209 (41%). The mean number of FT was greater (p < 0.001) among urban residents, women and those with a higher level of education. Only 1% had a CPITN of 0; 6% exhibited bleeding, 40% had calculus, 43% shallow and 10% deep pockets. Illiterate subjects were more likely to have DT (OR = 1.6, 95% CI = 1.4–1.8) and calculus or periodontal pockets (OR = 6.3, 95% CI = 5.1–7.8). Conclusion: The findings indicate an impaired oral health status in Iranian adults, particularly those of low social status and educational level.
Experimental Comparison of Bovine-Derived Xenograft, Xenograft-Autologous Bone Marrow and Autogenous Bone Graft for the Treatment of Bony Defects in the Rabbit Ulna
Authors: Keskin Davut , Gündoğdu Cemal , Atac Alpaslan Cevdet
Published: June 2007

Objectives: The purpose of this study was to determine whether bone marrow grafted percutanously has an effect on the healing of bony defects filled with bone-derived xenografts. Materials and Methods: Eighty New Zealand albino rabbits with anaverage age of approximately 1 year were divided into 4 groups, each consisting of 20 animals. Bony defect was induced in the ulnas of all rabbits by excising a 1-cm-long bone segment from the 3-cm proximal segment of the right distal radioulnar joint. Bone defects were treated simultaneously with bovine-derived xenograft, a combination of xenograft and bone marrow or on the 5th day following the filling of the segment with the xenograft and autogenous bone graft. Treatment results obtained for each application type were compared with each other with regard to the radiological, biochemical and histological criteria. Results: No significant statistical differences were determined between the groups in their 15th-day radiographs. The group treated with only xenograft from the first month onwards presented with the worst results and was significantly different from the other groups with respect to all evaluation criteria. No statistically significant difference was determined between rabbits treated with xenograft and bone marrow combination and with autogenous bone graft. Conclusions: This study revealed that when xenografts are combined with autologous bone marrow, their incorporation into the host bed accelerates significantly.
Using Intravascular Autoradiography for an Estimation of Proliferative Activity of Rat Mesenteric Microvessel Endothelial Cells during the First Month of Postnatal Development
Authors: Grigorieva T.A. , Belichenko V.M. , Aizman R.I. , Shoshenko C.A.
Published: June 2007

Background/Methods: We applied a novel method for studying endothelial cells (EC) by using autoradiography of cells labeled by 3H-thymidine: photo emulsion was administered into the vascular bed. In the flat transparent organ, this method allows to compare the mitotic activity of endothelium (MAE) depending on vessel diameter and animal age. Results: The number of the labeled ECs in animals was found to be almost equal in all vessels at the same age. The amount of labeled vessels and the density of the labeled nuclei increase towards near-capillary vessels of a diameter ≤10 µm. With age, ÐœÐE decreases in all categories of vessels. However, in 12-day-old animals, ÐœÐE temporarily increases. It was noted that high ÐœÐE in the bed of these rats precedes or coincides with the period of accelerated weight gain of the digestive system supplied by an increase in organ blood flow. Conclusions: By using the endothelial autoradiography method that we developed, we obtained the following evidence: (1) the number of ECs in the synthetic phase of the mitotic cycle in mesenterial vessels is approximately identical in animals of the same age, and (2) this number decreases with age, except during the period of accelerated organ growth, when MAE increases.
Cigarette Smoking and Incident Chronic Kidney Disease: A Systematic Review
Authors: Jones-Burton Charlotte , Seliger Stephen L. , Scherer Roberta W. , Mishra Shiraz I. , Vessal Ghazal , Brown Jeanine , Weir Matthew R. , Fink Jeffrey C.
Published: June 2007

Background: Several studies have examined the role of cigarette smoking in the development of renal disease in human populations. However, there have been no systematic reviews on the evidence linking smoking with incident renal disease. Methods: We performed an evidence-based evaluation of peer-reviewed research published during 1966–2005, from a search of five databases, including Ovid MEDLINE and EMBASE. Results: Of the 28 studies that were reviewed, 11 were excluded from the final analysis due to poor methodological quality (n = 6), no reported risk estimate for the association between smoking and kidney disease (n = 3), inability to find a Japanese translator (n = 1), and duplicate cohort (n = 1). Seventeen studies were included in the final analysis; seven studies found an overall significant association between smoking and incident chronic kidney disease, and three studies found a significantly increased risk of chronic kidney disease in current smokers that was gender and/or dose related. An increased risk of developing chronic kidney disease among smokers was significantly associated with male gender (relative risk 2.4, 95% confidence interval 1.2–4.5), >20 cigarettes smoked/day (odds ratio 1.51, 95% confidence interval 1.06–2.15, and relative risk 2.3, 95% confidence interval 1.2–4.3), and smoking >40 years (odds ratio 1.45, 95% confidence interval 1.00–2.09). A pooled estimate of the relative risk (meta-analysis) was deemed inappropriate due to the heterogeneity in methodologies utilized by the different studies. Conclusions: This comprehensive review reveals overall evidence for current cigarette smoking as a risk factor for incident chronic kidney disease. Further investigation is needed to more carefully examine the strength of the association between cigarette smoking and incident kidney disease.
Management of Superficial Vein Thrombosis and Thrombophlebitis: Status and Expert Opinion Document
Authors: Cesarone M.R. , Belcaro G. , Agus G. , Georgiev M. , Errichi B.M. , Marinucci R. , Errichi S. , Filippini A. , Pellegrini L. , Ledda A. , Vinciguerra G. , Ricci A. , Cipollone G. , Lania M. , Gizzi G. , Ippolito E. , Bavera P. , Fano F. , Dugall M. , Adovasio R. , Gallione L. , Del Boccio G. , Cornelli U. , Steigerwalt R. , Acerbi G. , Cacchio M. , Di Renzo A. , Hosoi M. , Stuard S. , Corsi M. , Di Ciano L. , Simeone E. , Collevecchio G. , Grossi M.G. , Di Giambattista F. , Carestia F. , Zukowski A.
Published: June 2007

Superficial vein thrombosis is characterized by clotting of superficial veins (ie, following direct trauma) with minimal inflammatory components. Superficial thrombophlebitis is a minimally thrombotic process of superficial veins associated with inflammatory changes and/or infection. Treatments generally include analgesics, elastic compression, anti-inflammatory agents, exercise and ambulation, and, in some cases, local or systemic anticoagulants. It is better to avoid bed rest and reduced mobility. Topical analgesia with nonsteroidal, anti-inflammatory creams applied locally to the superficial vein thrombosis/superficial thrombophlebitis area controls symptoms. Hirudoid cream (heparinoid) shortens the duration of signs/symptoms. Locally acting anticoagulants/antithrombotics (Viatromb®, Lipohep ®, spray Na-heparin) have positive effects on pain and on the reduction in thrombus size. Intravenous catheters should be changed every 24 to 48 hours (depending on venous flow and clinical parameters) to prevent superficial vein thrombosis/superficial thrombophlebitis and removed in case of events. Low molecular weight heparin prophylaxis and nitroglycerin patches distal to peripheral lines may reduce the incidence of superficial vein thrombosis/superficial thrombophlebitis in patients with vein catheters. In case of superficial vein thrombosis/superficial thrombophlebitis, vein lines should be removed. In neoplastic diseases and hematological disorders, anticoagulants may be necessary. Exercise reduces pain and the possibility of deep vein thrombosis. Only in cases in which pain is very severe is bed rest necessary. Deep vein thrombosis prophylaxis should be established in patients with reduced mobility. Antibiotics usually do not have a place in superficial vein thrombosis/superficial thrombophlebitis unless there are documented infections. Prevention of superficial vein thrombosis should be considered on the basis of patient's history and clinical evaluation.
Topical Heparin: New Observations
Authors: Cesarone M.R. , Belcaro G. , Errichi S. , Cornelli U. , Pellegrini L. , Ruffini I. , Errichi B.M. , Ledda A. , Vinciguerra G. , Ricci A. , Gizzi G. , Ippolito E. , Fano F. , Dugall M. , Cacchio M. , Di Renzo A. , Hosoi M. , Grossi M.G. , Stuard S. , Corsi M.
Published: June 2007

Topical effects of heparins on the skin need deeper investigations. The lack of evidence is mainly due to the lack of large investments in this field. Three main local actions of heparin on the skin can be defined: (a) the anticoagulant action, (b) the microcirculatory-modulatory action determining important control of the microcirculation in case of excessive vasoconstriction or vasodilatation, and (c) the `facilitatory action' on skin permeability allowing other drugs to diffuse better and faster into the skin (producing a therapeutic effect). These aspects have to be evaluated more extensively in both experimental and clinical conditions. Recent experimental studies demonstrate these effects of locally applied heparin. Therefore, key questions on local heparin administration such as skin penetration and the action on the local thrombi have promising answers. These observations suggest important clinical applications for local liposomal heparin. Both the potentials of local applications of heparin, particularly with new formulations, and some new aspects in the management of superficial vein thrombosis (SVT) can focus on locally applied heparin. SVT is an important clinical condition considering its frequency and the potentially heavy use of local heparin in this clinical problem. Results from new studies and observations presented in this issue of Angiology could be a window for suggesting new significant clinical applications and therapeutic solutions.