bluuebunny/crossref_metadata_2025 huggingface dataset:
In vivo, Noninvasive, Label-Free Detection and Eradication of Circulating Metastatic Melanoma Cells Using Two-Color Photoacoustic Flow Cytometry with a Diode Laser
Authors: Galanzha Ekaterina I. , Shashkov Evgeny V. , Spring Paul M. , Suen James Y. , Zharov Vladimir P.
Published: October 2009

The circulating tumor cell (CTC) count has been shown as a prognostic marker for metastasis development. However, its clinical utility for metastasis prevention remains unclear, because metastases may already be present at the time of initial diagnosis with existing assays. Their sensitivity ex vivo is limited by a small blood sample volume, whereas in vivo examination of larger blood volumes may be clinically restricted by the toxicity of labels used for targeting of CTCs. We introduce a method for in vivo photoacoustic blood cancer testing with a high-pulse-repetition-rate diode laser that, when applied to melanoma, is free of this limitation. It uses the overexpression of melanin clusters as intrinsic, spectrally-specific cancer markers and signal amplifiers, thus providing higher photoacoustic contrast of melanoma cells compared with a blood background. In tumor-bearing mouse models and melanoma-spiked human blood samples, we showed a sensitivity level of 1 CTC/mL with the potential to improve this sensitivity 103-fold in humans in vivo, which is impossible with existing assays. Additional advances of this platform include decreased background signals from blood through changes in its oxygenation, osmolarity, and hematocrit within physiologic norms, assessment of CTCs in deep vessels, in vivo CTC enrichment, and photoacoustic-guided photothermal ablation of CTCs in the bloodstream. These advances make feasible the early diagnosis of melanoma during the initial parallel progression of primary tumor and CTCs, and laser blood purging using noninvasive or hemodialysis-like schematics for the prevention of metastasis. [Cancer Res 2009;69(20):7926–34]
The Novel Tubulin-Targeting Agent Pyrrolo-1,5-Benzoxazepine-15 Induces Apoptosis in Poor Prognostic Subgroups of Chronic Lymphocytic Leukemia
Authors: McElligott Anthony M. , Maginn Elaina N. , Greene Lisa M. , McGuckin Siobhan , Hayat Amjad , Browne Paul V. , Butini Stefania , Campiani Giuseppe , Catherwood Mark A. , Vandenberghe Elisabeth , Williams D. Clive , Zisterer Daniela M. , Lawler Mark
Published: October 2009

Pyrrolo-1,5-benzoxazepine-15 (PBOX-15) is a novel microtubule depolymerization agent that induces cell cycle arrest and subsequent apoptosis in a number of cancer cell lines. Chronic lymphocytic leukemia (CLL) is characterized by clonal expansion of predominately nonproliferating mature B cells. Here, we present data suggesting PBOX-15 is a potential therapeutic agent for CLL. We show activity of PBOX-15 in samples taken from a cohort of CLL patients (n = 55) representing both high-risk and low-risk disease. PBOX-15 exhibited cytotoxicity in CLL cells (n = 19) in a dose-dependent manner, with mean IC50 of 0.55 μmol/L. PBOX-15 significantly induced apoptosis in CLL cells (n = 46) including cells with poor prognostic markers: unmutated IgVH genes, CD38 and zeta-associated protein 70 (ZAP-70) expression, and fludarabine-resistant cells with chromosomal deletions in 17p. In addition, PBOX-15 was more potent than fludarabine in inducing apoptosis in fludarabine-sensitive cells. Pharmacologic inhibition and small interfering RNA knockdown of caspase-8 significantly inhibited PBOX-15–induced apoptosis. Pharmacologic inhibition of c-jun NH2-terminal kinase inhibited PBOX-15–induced apoptosis in mutated IgVH and ZAP-70− CLL cells but not in unmutated IgVH and ZAP-70+ cells. PBOX-15 exhibited selective cytotoxicity in CLL cells compared with normal hematopoietic cells. Our data suggest that PBOX-15 represents a novel class of agents that are toxic toward both high-risk and low-risk CLL cells. The need for novel treatments is acute in CLL, especially for the subgroup of patients with poor clinical outcome and drug-resistant disease. This study identifies a novel agent with significant clinical potential. [Cancer Res 2009;69(21):8366–75]
The Cell Death–Inducing Activity of the Peptide Containing Noxa Mitochondrial-Targeting Domain Is Associated with Calcium Release
Authors: Seo Young-Woo , Woo Ha-Na , Piya Sujan , Moon Ae Ran , Oh Jae-Wook , Yun Cheol-Won , Kim Kyung-Keun , Min Ji-Young , Jeong Seon-Yong , Chung Seyung , Song Peter I. , Jeong Seong-Yun , Choi Eun Kyung , Seol Dai-Wu , Kim Tae-Hyoung
Published: October 2009

DNA damage stabilizes the p53 tumor suppressor protein that determines the cell fate by either cell cycle arrest or cell death induction. Noxa, the BH3-only Bcl-2 family protein, was shown to be a key player in p53-induced cell death through the mitochondrial dysfunction; however, the molecular mechanism by which Noxa induces the mitochondrial dysfunction to cause cell death in response to genotoxic agents is largely unknown. Here, we show that the mitochondrial-targeting domain (MTD) of Noxa is a prodeath domain. Peptide containing MTD causes massive necrosis in vitro through cytosolic calcium increase; it is released from the mitochondria by opening the mitochondrial permeability transition pore. MTD peptide–induced cell death can be inhibited by calcium chelator BAPTA-AM. Moreover, MTD peptide shows the potent tumor-killing activities in mice by joining with tumor-homing motifs. [Cancer Res 2009;69(21):8356–65]
Intravital Imaging Reveals Transient Changes in Pigment Production and Brn2 Expression during Metastatic Melanoma Dissemination
Authors: Pinner Sophie , Jordan Peter , Sharrock Kirsty , Bazley Laura , Collinson Lucy , Marais Richard , Bonvin Elise , Goding Colin , Sahai Erik
Published: October 2009

How melanoma acquire a metastatic phenotype is a key issue. One possible mechanism is that metastasis is driven by microenvironment-induced switching between noninvasive and invasive states. However, whether switching is a reversible or hierarchical process is not known and is difficult to assess by comparison of primary and metastatic tumors. We address this issue in a model of melanoma metastasis using a novel intravital imaging method for melanosomes combined with a reporter construct in which the Brn-2 promoter drives green fluorescent protein (GFP) expression. A subpopulation of cells containing little or no pigment and high levels of Brn2::GFP expression are motile in the primary tumor and enter the vasculature. Significantly, the less differentiated state of motile and intravasated cells is not maintained at secondary sites, implying switching between states as melanoma cells metastasize. We show that melanoma cells can switch in both directions between high- and low-pigment states. However, switching from Brn2::GFP high to low was greatly favored over the reverse direction. Microarray analysis of high- and low-pigment populations revealed that transforming growth factor (TGF)β2 was up-regulated in the poorly pigmented cells. Furthermore, TGFβ signaling induced hypopigmentation and increased cell motility. Thus, a subset of less differentiated cells exits the primary tumor but subsequently give rise to metastases that include a range of more differentiated and pigment-producing cells. These data show reversible phenotype switching during melanoma metastasis. [Cancer Res 2009;69(20):7969–77]
Kupffer Cell Suppression of CD8+ T Cells in Human Hepatocellular Carcinoma Is Mediated by B7-H1/Programmed Death-1 Interactions
Authors: Wu Ke , Kryczek Ilona , Chen Lieping , Zou Weiping , Welling Theodore H.
Published: October 2009

B7-H1 is a recently identified B7 family member that, along with one of its receptors, programmed death-1 (PD-1), has been involved in multiple immunopathologic scenarios. However, the nature of B7-H1 and PD-1 in human hepatocellular carcinoma (HCC) remains poorly defined. We investigated the expression and functional relevance of this pathway in patients with HCC. We showed that B7-H1 expression on Kupffer cells (KC) was increased in tumor tissues compared with surrounding nontumor liver tissues in patients with HCC and this correlated with poorer survival. Coculture of HCC cells with monocytes showed that tumor-associated interleukin-10 contributed to the induction of B7-H1 in the HCC environment. We further observed that the levels of PD-1+CD8+ T cells were higher in tumor tissues than in nontumor tissues. B7-H1+ KCs and PD-1+ T cells were colocalized in the HCC stroma. PD-1+CD8+ T cells had decreased proliferative ability and effector function as shown by reduced granule and cytokine expression compared with PD-1− T cells. Importantly, blocking KC B7-H1 interaction with PD-1+CD8+ cells using neutralizing antibodies recovered effector T-cell function. Our data indicate that the B7-H1/PD-1 axis contributes to immune suppression in human HCC, with blockade of this pathway carrying important therapeutic implications. [Cancer Res 2009;69(20):8067–75]
Progressive Chromatin Repression and Promoter Methylation of CTNNA1 Associated with Advanced Myeloid Malignancies
Authors: Ye Ying , McDevitt Michael A. , Guo Mingzhou , Zhang Wei , Galm Oliver , Gore Steven D. , Karp Judith E. , Maciejewski Jaroslaw P. , Kowalski Jeanne , Tsai Hua-Ling , Gondek Lukasz P. , Tsai Hsing-Chen , Wang Xiaofei , Hooker Craig , Smith B. Douglas , Carraway Hetty E. , Herman James G.
Published: October 2009

Complete loss or deletion of the long arm of chromosome 5 is frequent in myelodysplastic syndrome (MDS) and acute myelogenous leukemia (AML). The putative gene(s) deleted and responsible for the pathogenesis of these poor prognosis hematologic disorders remain controversial. This study is a comprehensive analysis of previously implicated and novel genes for epigenetic inactivation in AML and MDS. In 146 AML cases, methylation of CTNNA1 was frequent, and more common in AML patients with 5q deletion (31%) than those without 5q deletion (14%), whereas no methylation of other 5q genes was observed. In 31 MDS cases, CTNNA1 methylation was only found in high-risk MDS (≥RAEB2), but not in low-risk MDS (<RAEB2), indicating that CTNNA1 methylation might be important in the transformation of MDS to AML. CTNNA1 expression was lowest in AML/MDS patients with CTNNA1 methylation, although reduced expression was found in some patients without promoter methylation. Repressive chromatin marks (H3K27me3) at the promoter were identified in CTNNA1-repressed AML cell lines and primary leukemias, with the most repressive state correlating with DNA methylation. These results suggest progressive, acquired epigenetic inactivation at CTNNA1, including histone modifications and promoter CpG methylation, as a component of leukemia progression in patients with both 5q− and non-5q– myeloid malignancies. [Cancer Res 2009;69(21):8482–90]
Molecular Consequences of Genetic Variations in the Glutathione Peroxidase 1 Selenoenzyme
Authors: Zhuo Pin , Goldberg Marci , Herman Lauren , Lee Bao-Shiang , Wang Hengbing , Brown Rhonda L. , Foster Charles B. , Peters Ulrike , Diamond Alan M.
Published: October 2009

Accumulating data have implicated the selenium-containing cytosolic glutathione peroxidase, GPx-1, as a determinant of cancer risk and a mediator of the chemopreventive properties of selenium. Genetic variants of GPx-1 have been shown to be associated with cancer risk for several types of malignancies. To investigate the relationship between GPx-1 enzyme activity and genotype, we measured GPx-1 enzyme activity and protein levels in human lymphocytes as a function of the presence of two common variations: a leucine/proline polymorphism at codon 198 and a variable number of alanine-repeat codons. Differences in GPx activity among these cell lines, as well as in the response to the low-level supplementation of the media with selenium, indicated that factors other than just genotype are significant in determining activity. To restrict the study to genotypic effects, human MCF-7 cells were engineered to exclusively express allelic variants representing a combination of either a codon 198 leucine or proline and either 5 or 7 alanine-repeat codons following transfection of GPx-1 expression constructs. Transfectants were selected and analyzed for GPx-1 enzyme activity and protein levels. GPx-1 with 5 alanines and a leucine at codon 198 showed a significantly higher induction when cells were incubated with selenium and showed a distinct pattern of thermal denaturation as compared with GPx-1 encoded by the other examined alleles. The collective data obtained using both lymphocytes and MCF-7 indicate that both intrinsic and extrinsic factors cooperate to ultimately determine the levels of this enzyme available to protect cells against DNA damage and mutagenesis. [Cancer Res 2009;69(20):8183–90]
Protein Kinase CK2 Regulates Cytoskeletal Reorganization during Ionizing Radiation–Induced Senescence of Human Mesenchymal Stem Cells
Authors: Wang Daojing , Jang Deok-Jin
Published: October 2009

Human mesenchymal stem cells (hMSC) are critical for tissue regeneration. How hMSC respond to genotoxic stresses and potentially contribute to aging and cancer remain underexplored. We showed that ionizing radiation induced cellular senescence of hMSC over a period of 10 days, showing a critical transition between days 3 and 6. This was confirmed by senescence-associated β-galactosidase staining, protein expression profiles of key cell cycle regulators (retinoblastoma protein, p53, p21waf1/Cip1, and p16INK4A), and senescence-associated secretory phenotypes (interleukin-8, interleukin-12, GRO, and MDC). We observed dramatic cytoskeletal reorganization of hMSC through reduction of myosin-10, redistribution of myosin-9, and secretion of profilin-1. Using a SILAC-based phosphoproteomics method, we detected significant reduction of myosin-9 phosphorylation at Ser1943, coinciding with its redistribution. Importantly, through treatment with cell-permeable inhibitors (4,5,6,7-tetrabromo-1H-benzotriazole and 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole) and gene knockdown using RNA interference, we identified CK2, a kinase responsible for myosin-9 phosphorylation at Ser1943, as a key factor contributing to the radiation-induced senescence of hMSC. We showed that individual knockdown of CK2 catalytic subunits CK2α and CK2α′ induced hMSC senescence. However, only knockdown of CK2α resulted in morphologic phenotypes resembling those of radiation-induced senescence. These results suggest that CK2α and CK2α′ play differential roles in hMSC senescence progression, and their relative expression might represent a novel regulatory mechanism for CK2 activity. [Cancer Res 2009;69(20):8200–7]
Direct Evidence that Bevacizumab, an Anti-VEGF Antibody, Up-regulates SDF1α, CXCR4, CXCL6, and Neuropilin 1 in Tumors from Patients with Rectal Cancer
Authors: Xu Lei , Duda Dan G. , di Tomaso Emmanuelle , Ancukiewicz Marek , Chung Daniel C. , Lauwers Gregory Y. , Samuel Rekha , Shellito Paul , Czito Brian G. , Lin Pei-Chun , Poleski Martin , Bentley Rex , Clark Jeffrey W. , Willett Christopher G. , Jain Rakesh K.
Published: October 2009

Clinical studies converge on the observation that circulating cytokines are elevated in most cancer patients by anti–vascular endothelial growth factor (VEGF) therapy. However, the source of these molecules and their relevance in tumor escape remain unknown. We examined the gene expression profiles of cancer cells and tumor-associated macrophages in tumor biopsies before and 12 days after monotherapy with the anti-VEGF antibody bevacizumab in patients with rectal carcinoma. Bevacizumab up-regulated stromal cell–derived factor 1α (SDF1α), its receptor CXCR4, and CXCL6, and down-regulated PlGF, Ang1, and Ang2 in cancer cells. In addition, bevacizumab decreased Ang1 and induced neuropilin 1 (NRP1) expression in tumor-associated macrophages. Higher SDF1α plasma levels during bevacizumab treatment significantly associated with distant metastasis at three years. These data show that VEGF blockade up-regulates inflammatory pathways and NRP1, which should be evaluated as potential targets for improving anti-VEGF therapy. [Cancer Res 2009;69(20):7905–10]
HDAC Inhibitor SNDX-275 Induces Apoptosis in erbB2-Overexpressing Breast Cancer Cells via Down-regulation of erbB3 Expression
Authors: Huang Xiaoping , Gao Lizhi , Wang Shuiliang , Lee Choon-Kee , Ordentlich Peter , Liu Bolin
Published: October 2009

Breast cancer is a highly heterogeneous disease with distinct histologic subtypes. Targeted therapies such as endocrine therapy and growth factor receptor inhibitors have had a significant impact on the treatment of metastatic breast cancer patients. Unfortunately, resistance to these agents eventually occurs, and currently represents a significant clinical problem in the management of breast cancers. Inhibitors of histone deacetylases (HDACi) exhibit anticancer activity in a variety of tumor cell models and have been shown to target mechanisms of resistance to a number of targeted agents. It is unclear, however, if there are specific breast cancer subtypes for which an HDACi may be more or less effective. Here, we report that the class I isoform–selective HDACi entinostat (SNDX-275) preferentially inhibits cell proliferation/survival and inactivates downstream signaling in erbB2-overexpressing compared with basal breast cancer cells. SNDX-275 reduces the levels of both erbB2 and erbB3, as well as significantly decreases P-erbB2, P-erbB3, P-Akt, and P-MAPK in erbB2-overexpressing cells. Additionally, SNDX-275 promotes apoptosis and induces cell cycle arrest predominantly at G1 phase in erbB2-overexpressing cells, whereas SNDX-275 mainly induces G2-M arrest in basal breast cancer cells. The cellular bias of SNDX-275 is shown to be related partly to the levels of erbB3 expression that directly impact the ability of SNDX-275 to inhibit proliferation/survival of the erbB2-overexpressing breast cancer cells. These findings show that SNDX-275 may be developed as a novel therapeutic agent to treat breast cancers with coexpression of both erbB2 and erbB3. [Cancer Res 2009;69(21):8403–11]
SC68896, a Novel Small Molecule Proteasome Inhibitor, Exerts Antiglioma Activity In vitro and In vivo
Authors: Roth Patrick , Kissel Maria , Herrmann Caroline , Eisele Günter , Leban Johann , Weller Michael , Schmidt Friederike
Published: October 2009

Purpose: Glioblastomas are among the most lethal neoplasms, with a median survival of <1 year. Modulation of the proteasome function has emerged as a novel approach to cancer pharmacotherapy. Here, we characterized the antitumor properties of SC68896, a novel small molecule proteasome inhibitor.

Experimental Design: Different tumor cell lines were tested by crystal violet staining for sensitivity to SC68896, given alone or in combination with death ligands. The molecular mechanisms mediating SC68896-induced cell death and changes in cell cycle progression were assessed by immunoblot and flow cytometry. An orthotopic human glioma xenograft model in nude mice was used to examine the in vivo activity of SC68896.

Results: SC68896 inhibits the proliferation of cell lines of different types of cancer, including malignant glioma. Exposure of LNT-229 glioma cells to SC68896 results in a concentration- and time-dependent inhibition of the proteasome, with a consequent accumulation of p21 and p27 proteins, cell cycle arrest, caspase cleavage, and induction of apoptosis. Using RNA interference, we show that the effect of SC68896 on glioma cells is facilitated by wild-type p53. SC68896 sensitizes glioma cells to tumor necrosis factor–related apoptosis–inducing ligand and CD95 ligand and up-regulates the cell surface expression of the tumor necrosis factor–related apoptosis–inducing ligand receptor cell death receptors 4 and 5, which may contribute to this sensitization. Intracerebral glioma–bearing nude mice treated either i.p. or intratumorally with SC68896 experience prolonged survival.

Conclusions: SC68896 is the first proteasome inhibitor that exerts antiglioma activity in vivo. It may represent a novel prototype agent for the treatment of malignant gliomas and warrants clinical evaluation. (Clin Cancer Res 2009;15(21):6609–18)
Mammary-Derived Growth Inhibitor Alters Traffic of EGFR and Induces a Novel Form of Cetuximab Resistance
Authors: Nevo Jonna , Mattila Elina , Pellinen Teijo , Yamamoto Daniel L. , Sara Henri , Iljin Kristiina , Kallioniemi Olli , Bono Petri , Heikkilä Päivi , Joensuu Heikki , Wärri Anni , Ivaska Johanna
Published: October 2009

Purpose: Only few predictive factors for the clinical activity of anti–epidermal growth factor receptor (EGFR) therapy are available. Mammary-derived growth inhibitor (MDGI) is a small cytosolic protein suggested to play a role in the differentiation of epithelial cells. Here, we have investigated the effect of MDGI expression on the EGFR signaling and cetuximab responsiveness of cancer cells.

Experimental Design: MDGI mRNA expression was investigated in clinical breast and lung cancer samples and in nontransformed and malignant cell lines. The effect of ectopic expression of MDGI on EGFR, ErbB2, and integrin function and traffic was investigated in breast and lung cancer cell lines using multiple methods. The effect of anti-EGFR agents on these cells were tested by cell proliferation measurements and by assessing tumor growth of breast cancer cells in cetuximab treated and control athymic nude mice.

Results: Here, we show that although MDGI is absent in cultured cell lines because of epigenetic silencing, MDGI mRNA is expressed in 40% of clinical breast carcinomas and 85% of lung cancers. Ectopic expression of MDGI rendered breast and lung cancer cells resistant to the anti-EGFR antibody cetuximab in vitro and in an orthotopic breast cancer xenograft model in vivo. When expressed in cancer cells, MDGI induces intracellular accumulation of EGFR, but not ErbB2, and the internalized receptor is phosphorylated and not degraded.

Conclusions: MDGI-driven inherent desensitization of cancer cells is a novel molecular mechanism for resistance to the anti-EGFR antibody therapy, and MDGI may be a biomarker for responsiveness to anti-EGFR antibody therapy.(Clin Cancer Res 2009;15(21):6570–81)
Recombinant Peptides as Biomarkers for Tumor Response to Molecular Targeted Therapy
Authors: Passarella Ralph J. , Zhou Li , Phillips John G. , Wu Hongmei , Hallahan Dennis E. , Diaz Roberto
Published: October 2009

Purpose: Phage display technology can be used to identify peptide sequences that bind rapidly and specifically to tumors responding to sunitinib therapy. These peptides may help to address problems with current methods of assessing tumor response to therapy that can be slow and have limited usage.

Experimental Design: The peptide of interest was isolated after four rounds of biopanning in MDA-MB-231 and MCF-7 xenografted tumors. The binding location of the peptide was investigated with immunohistochemistry. Its in vivo ability to bind to breast tumors responding to therapy was determined by treating nude mice, xenografted with various tumor cell lines, with sunitinib and using near IR imaging to assess the ability of the peptide conjugated to Alexafluor-750 to bind tumors.

Results: EGEVGLG was the dominant sequence isolated from biopanning. This peptide showed increased binding relative to control groups in two cancer cell lines (MDA-MB-435 and MCF-7 human breast) responding to sunitinib treatment, whereas no elevated binding occurred in vitro when samples were incubated with tumor cells that are unresponsive to sunitinib treatment (B16 melanoma and BxPC3 pancreatic). Mice xenografted with tumors that are responsive to sunitinib therapy showed increased peptide binding when compared with untreated control. Mice bearing tumors unresponsive to sunitinib therapy showed no increased peptide binding between treated and untreated groups.

Conclusion: The use of recombinant peptides to assess the pharmacodynamic response of cancer holds promise in minimizing the duration of ineffective treatment regimens in patients, potentially providing a more rapid and less invasive assessment of cancer response to systemic therapy. (Clin Cancer Res 2009;15(20):6421–9)
Clinical Predictors and Algorithm for the Genetic Diagnosis of Pheochromocytoma Patients
Authors: Erlic Zoran , Rybicki Lisa , Peczkowska Mariola , Golcher Henriette , Kann Peter H. , Brauckhoff Michael , Müssig Karsten , Muresan Michaela , Schäffler Andreas , Reisch Nicole , Schott Matthias , Fassnacht Martin , Opocher Giuseppe , Klose Silke , Fottner Christian , Forrer Flavio , Plöckinger Ursula , Petersenn Stephan , Zabolotny Dimitry , Kollukch Oleg , Yaremchuk Svetlana , Januszewicz Andrzej , Walz Martin K. , Eng Charis , Neumann Hartmut P.H. ,   for the European-American Pheochromocytoma Study Group
Published: October 2009

Purpose: Six pheochromocytoma susceptibility genes causing distinct syndromes have been identified; approximately one of three of all pheochromocytoma patients carry a predisposing germline mutation. When four major genes (VHL, RET, SDHB, SDHD) are analyzed in a clinical laboratory, costs are ∼$3,400 per patient. The aim of the study is to systematically obtain a robust algorithm to identify who should be genetically tested, and to determine the order in which genes should be tested.

Experimental Design: DNA from 989 apparently nonsyndromic patients were scanned for germline mutations in the genes VHL, RET, SDHB, SDHC, and SDHD. Clinical parameters were analyzed as potential predictors for finding mutations by multiple logistic regression, validated by bootstrapping. Cost reduction was calculated between prioritized gene testing compared with that for all genes.

Results: Of 989 apparently nonsyndromic pheochromocytoma cases, 187 (19%) harbored germline mutations. Predictors for presence of mutation are age <45 years, multiple pheochromocytoma, extra-adrenal location, and previous head and neck paraganglioma. If we used the presence of any one predictor as indicative of proceeding with gene testing, then 342 (34.6%) patients would be excluded, and only 8 carriers (4.3%) would be missed. We were also able to statistically model the priority of genes to be tested given certain clinical features. E.g., for patients with prior head and neck paraganglioma, the priority would be SDHD>SDHB>RET>VHL. Using the clinical predictor algorithm to prioritize gene testing and order, a 44.7% cost reduction in diagnostic process can be achieved.

Conclusions: Clinical parameters can predict for mutation carriers and help prioritize gene testing to reduce costs in nonsyndromic pheochromocytoma presentations. (Clin Cancer Res 2009;15(20):6378–85)
Tumor-Expressed B7-H1 and B7-DC in Relation to PD-1+ T-Cell Infiltration and Survival of Patients with Cervical Carcinoma
Authors: Karim Rezaul , Jordanova Ekaterina S. , Piersma Sytse J. , Kenter Gemma G. , Chen Lieping , Boer Judith M. , Melief Cornelis J. M. , van der Burg Sjoerd H.
Published: October 2009

Purpose: The interaction between programmed cell death 1 (PD-1), expressed by activated effector or regulatory T cells, and B7-H1 (PD-L1) and B7-DC (PD-L2) results in the inhibition of T-cell function. The aim of this study was to determine B7-H1, B7-DC, and PD-1 expression in cervical carcinoma.

Experimental Design: A tissue microarray of a well-defined group of 115 patients was stained with antibodies against B7-H1 and B7-DC. Three-color fluorescent immunohistochemistry was used to study the number and phenotype of tumor-infiltrating T cells expressing PD-1. Additional analyses consisted of in vitro T-cell suppression assays.

Results: B7-H1 was expressed in 19%, and B7-DC was expressed by 29% of the 115 tumors. PD-1 was expressed by more than half of both the infiltrating CD8+ T cells and CD4+Foxp3+ T cells, irrespective of B7-H1 or B7-DC expression by tumors. The expression of B7-H1 did not show a direct impact on patient survival. However, subgroup analysis revealed that patients with a relative excess of infiltrating regulatory T cells displayed a better survival when the tumor was B7-H1 positive (P = 0.033). Additional studies showed that the presence of B7-H1 during the activation of CD4+Foxp3+ regulatory T cells impaired their suppressive function in a functional in vitro assay.

Conclusions: B7-H1 is expressed on only a minority of cervical cancers and does not influence the survival of patients with cervical cancer. PD-1 is expressed by a vast number of infiltrating CD8 T cells, suggesting that blocking of PD-1 could have therapeutic potential in cervical cancer patients. (Clin Cancer Res 2009;15(20):6341–7)
Phenotypic Knockout of CXCR4 by a Novel Recombinant Protein TAT/54R/KDEL Inhibits Tumors Metastasis
Authors: Ma Wei-Feng , Du Jun , Fu Liu-Peng , Fang Rui , Chen Hong-Yuan , Cai Shao-Hui
Published: October 2009

The chemokine receptor, CXCR4, and its specific ligand, CXCL12, have been proven to regulate the directional trafficking and invasion of breast cancer cells to sites of metastases, and similar phenomena have also been identified in many malignant tumors that aberrantly overexpress CXCR4. Therefore, blocking the interaction between CXCR4 and CXCL12 is considered a possible approach to efficiently prevent cancer metastasis. Employing a cellular phenotypic knockout strategy based on intrakines, we developed a novel recombinant chimeric protein, TAT/54R/KDEL, which contains three distinct functional domains: CXCL12/54R, a mutant of CXCL12 with CXCR4 antagonism, as well as HIV-derived TAT (47-57) and an endoplasmic reticulum retention four-peptide sequence KDEL that links at its NH2 and COOH termini, respectively. Using the MOLT-4 cell line, which expressed CXCR4 highly and stably in vitro, we determined that TAT/54R/KDEL was able to efficiently transfer into the endoplasmic reticulum of tumor cells, where it specifically binds to the newly synthesized CXCR4 and prevents the latter from reaching the surface. Chemotaxis assays showed that the cells treated with TAT/54R/KDEL failed to migrate toward CXCL12. Furthermore, we observed that the systemic treatment of TAT/54R/KDEL could impair lung metastasis in a highly metastatic mammary cancer cell line, 4T1 cells, with the decrease of CXCR4 on their membrane. Our results suggest that the phenotypic knockout strategy of CXCR4 using a novel recombinant protein TAT/54R/KDEL might be a possible approach for inhibiting relative tumor metastasis mediated by CXCR4/CXCL12 interaction. (Mol Cancer Res 2009;7(10):1613–21)
Mutant p53 (G199V) Gains Antiapoptotic Function through Signal Transducer and Activator of Transcription 3 in Anaplastic Thyroid Cancer Cells
Authors: Kim Tae-Hyun , Lee Sang Yull , Rho Jee Hyun , Jeong Na Young , Soung Young Hwa , Jo Wol Soon , Kang Do-Young , Kim Sung-Heun , Yoo Young Hyun
Published: October 2009

In the present study, we identified a missense mutation (G199V) in KAT-18 cell line established from primary cultures of anaplastic thyroid cancer (ATC). Notably, knockdown of this mutant (mt) p53 reduced cell viability and exerted antitumor activity equivalent to high doses of several chemotherapeutic agents. We showed that p53 knockdown had an antitumor effect via the induction of apoptosis. We further examined the underlying mechanism by which mt p53 (G199V) gains antiapoptotic function in KAT-18 cells. Microarray analysis revealed that p53 knockdown modified the expression of numerous apoptosis-related genes. Importantly, p53 knockdown led to downregulation of signal transducer and activator of transcription-3 (STAT3) gene expression. We further observed that p53 knockdown induced the downregulation of STAT3 protein. We also observed that a STAT3 inhibitor augmented the reduction of cell viability induced by p53 knockdown, whereas interleukin-6 treatment alleviated this effect. In addition, overexpression of STAT3 protected ATC cells against cell death induced by p53 knockdown. Taken together, these data show that mt p53 (G199V) gains antiapoptotic function mediated by STAT3 in ATC cells. Inhibition of the function of mt p53 (G199V) could be a novel and useful therapeutic strategy for decreasing the extent and severity of toxicity due to chemotherapeutic agents. (Mol Cancer Res 2009;7(10):1645–54)
Galectin-3 Targeted Therapy with a Small Molecule Inhibitor Activates Apoptosis and Enhances Both Chemosensitivity and Radiosensitivity in Papillary Thyroid Cancer
Authors: Lin Chi-Iou , Whang Edward E. , Donner David B. , Jiang Xiaofeng , Price Brendan D. , Carothers Adelaide M. , Delaine Tamara , Leffler Hakon , Nilsson Ulf J. , Nose Vania , Moore, Francis D. , Ruan Daniel T.
Published: October 2009

Although most patients with papillary thyroid cancer (PTC) have favorable outcomes, some have advanced PTC that is refractory to external beam radiation and systemic chemotherapy. Galectin-3 (Gal-3) is a β-galactoside–binding protein with antiapoptotic activity that is consistently overexpressed in PTC. The purpose of this study is to determine if Gal-3 inhibition promotes apoptosis, chemosensitivity, and radiosensitivity in PTC. PTC cell lines (8505-C and TPC-1) and human ex vivo PTC were treated with a highly specific small molecule inhibitor of Gal-3 (Td131_1). Apoptotic activity was determined by flow cytometric analysis as well as caspase-3 and PARP cleavage. The minimum inhibitory concentrations of Td131_1 and doxorubicin were determined, and their combined effects were measured to test for synergistic activity. The effects of Td131_1 on radiosensitivity were determined by a clonogenic assay. Td131_1 promoted apoptosis, improved radiosensitivity, and synergistically enhanced chemosensitivity to doxorubicin in PTC cell lines. In PTC ex vivo, Td131_1 treatment alone induced the cleavage of caspase-3 and PARP. Td131_1 and doxorubicin together activated apoptosis in PTC ex vivo to a greater degree than their combined individual effects. Td131_1 activated apoptosis and had synergistic activity with doxorubicin in PTC. We conclude that Gal-3 targeted therapy is a promising therapeutic strategy for advanced PTC that is refractory to surgery and radioactive iodine therapy. (Mol Cancer Res 2009;7(10):1655–62)
Antegrade Mini-Invasive Nephroureterectomy: Laparoscopic Nephrectomy, Transurethral Excision of Ureterovesical Junction and Lower Abdominal Incision
Authors: Hora Milan , Eret Viktor , Ürge Tomáš , Klečka Jiří , Kočovská Petra , Petr Stránský , Hes Ondřej , Ferda Jiří
Published: October 2009

Introduction: We describe another variant of nephroureterectomy – antegrade mini-invasive nephroureterectomy (AMNUE). Methods: AMNUE starts with a laparoscopic nephrectomy in the flank position. The specimen is enclosed in a bag without dividing the ureter, and the patient is positioned to the lithotomy position. Then the ureterovesical junction is excised transurethrally with a Collins knife. Finally, the specimen is removed and the ureter is plucked out through a short lower abdomen incision. Patients: From March 2005 to November 2008, 35 patients underwent nephroureterectomy: 7 as an open procedure, 8 as a laparoscopic nephrectomy with open ureterectomy, 8 as a complete laparoscopic nephroureterectomy, and 12 were admitted into the AMNUE group (7 men and 5 women, mean age 71 ± 7 years, range 54–81 years). Results: Tumors were found 6 times on both sides. The mean operation time was 165 ± 32 min (105–210 min), and the mean blood loss was 150 ± 91 ml (50–400 ml). Histology revealed 11 urothelial cancers and 1 papillary renal cell carcinoma. There was only 1 hematoma of the abdominal wall. Conclusion: AMNUE is a fast, safe and easily reproducible technique. It eliminates the risk of spillage of tumorous cells into the urine, which is possible in techniques where the ureter is excised with a Collins knife as the first procedure. The disadvantages of this approach are the necessary repositioning of the patient and that the long-term oncological results are currently unknown. AMNUE can be used when a complete laparoscopic nephroureterectomy is not technically feasible due to problems in the pelvis.
Prognostic Significance of Hydronephrosis in Bladder Cancer Treated by Radical Cystectomy
Authors: Resorlu Berkan , Baltaci Sumer , Resorlu Mustafa , Ergun Gul , Abdulmajeed Mohammed , Haliloglu Ahmet Hakan , Gogus Cagatay , Beduk Yasar
Published: October 2009

Objectives: Hydronephrosis is a common finding in patients with bladder cancer. This study aims at an analysis of the association between hydronephrosis, pathologic characteristics of bladder cancer and cancer-specific survival rate after radical cystectomy. Methods: Clinical data of 241 patients who underwent radical cystectomy between 1990 and 2007 for invasive bladder cancer were analyzed retrospectively. The significance of the relation between the clinical and pathological findings were determined by the χ2 test. The Kaplan-Meier test was used for the estimation of disease-specific survival obtained according to hydronephrosis, and the differences were examined by the log-rank test. Multivariate analysis was performed by the Cox regression model. Results: Of 241 patients, 39 (16.2%) had unilateral and 13 (5.4%) bilateral hydronephrosis. Five-year cancer-specific survival rates were reported as 63.4% in non-hydronephrotic and 11.57% in hydronephrotic patients (log-rank test, p < 0.001). The presence of hydronephrosis was associated with advanced pT stage, higher tumor grade and lymph node metastases (p < 0.001, p < 0.001 and p < 0.001, respectively). Multivariate analysis showed that hydronephrosis is an important factor directly affecting cancer-specific survival (0.0264). Conclusions: The presence of hydronephrosis prior to radical cystectomy is a significant prognostic parameter, associated with poor cancer-specific survival and advanced disease stage.
Micropapillary Pattern in Urothelial Carcinoma: A Clinicopathological Analysis
Authors: Alkibay Turgut , Sözen Sinan , Gürocak Serhat , Işik Gönül İpek , Poyraz Aylar , Üre İyimser
Published: October 2009

Objectives: Our aim was to review our pathological archive to find out the actual incidence of micropapillary pattern (MPP) in our urothelial carcinoma patient population and determine its correlation with clinical outcome. Patients and Methods: 14 out of 170 cases with complete clinical follow-up were clinicopathologically analyzed. The extent of MPP was determined as tumor percentage. Results: 12 further cases with MPP were defined in the review. The percentage of patients with positive MPP increased in parallel to the tumor stage. There was no considerable difference between MPP-positive and MPP-negative groups according to the progression rates in non-muscle-invasive and muscle-invasive groups. Progression-free survival was much shorter in MPP cases, but again without statistical significance. Also, there was no significant relation between percentage of MPP and progression-free survival. Conclusions: Awareness of pathologists about MPP and its highly possible relation with aggressive behavior must be raised, as it may be more common than reported. A multicentric review of a large number of cases with MPP is needed for a better definition of its biological behavior. Focal MPP cases may have a better prognosis but this needs to be confirmed.
Preoperative Erythrocyte Sedimentation Rate Is an Independent Prognostic Factor in Japanese Patients with Localized Clear Cell Renal Cell Carcinoma
Authors: Kawai Yoshihisa , Matsuyama Hideyasu , Korenaga Yoshihito , Misumi Taku , Eguchi Satoshi , Hara Tomohiko , Nagao Kazuhiro , Ohmi Chietaka , Sakano Shigeru , Naito Katsusuke
Published: October 2009

Introduction: Few studies have examined the prognostic significance of common laboratory variables in Japanese patients with localized clear cell renal cell carcinoma (CCRCC). We evaluate the prognostic significance of preoperative laboratory variables in Japanese patients with localized CCRCC. Patients and Methods: The study included 110 Japanese patients who were pathologically confirmed as nonmetastatic CCRCC (pT1–3 N0M0) after radical nephrectomy at our institution. We assessed the clinical (including laboratory measurements) and pathological findings, with the survival rates after surgery. Results: Tumor stage and erythrocyte sedimentation rate (ESR) were identified as significant independent prognostic factors of progression-free survival in multivariate analysis. As for the prognostic factors for disease-specific survival, tumor stage and ESR had prognostic significance both in univariate and multivariate analyses. When the analysis was limited to pT1, multivariate analysis showed that only ESR was an independent prognostic factor for disease-specific survival. Conclusions: Preoperative ESR is an independent prognostic factor in Japanese patients with localized CCRCC, especially in patients with pT1.
Open-Label, Longitudinal Study of Tamsulosin for Functional Bladder Outlet Obstruction in Women
Authors: Costantini Elisabetta , Lazzeri Massimo , Bini Vittorio , Zucchi Alessandro , Fioretti Fabrizio , Frumenzio Emanuela , Porena Massimo
Published: October 2009

Objectives: To investigate the clinical effect of tamsulosin for the treatment of functional bladder outlet obstruction (BOO) in adult women. Methods: In a prospective, longitudinal open-label study, 63 women affected by functional BOO were treated with tamsulosin. Inclusion criteria were: age >18 years, reporting voiding symptoms (intermittent stream, hesitancy, straining and/or a feeling of incomplete emptying), uroflowmetry maximum flow rate (Qmax) under 12 ml and/or presence of postvoid residual greater than 50% of the voiding volume. Exclusion criteria were: patients suffering from any other anatomical or functional disorder such as urethral stenosis, pelvic organ prolapse, neurological disturbances, or systemic diseases that could impact upon bladder voiding. Tamsulosin was administered in a single daily dose of 0.4 mg for at least 30 days. Primary outcomes were clinical efficacy and Qmax improvement; secondary outcomes were tolerability and safety. Voiding and storage symptoms and uroflowmetry results were assessed before and at the end of the α-blocker therapy. Results: Tamsulosin therapy was well tolerated. After therapy voiding symptoms improved in 71.4% of patients (45/63; p < 0.0001), and if associated with storage symptoms in 66.67% (26/39; p < 0.00001). Recurrent infections were reduced by 50% in 81% (21/26) of patients. Uroflowmetry parameters improved in 36/63 patients (57.1%). Postvoiding residue improved in 62.5% (10/16) and disappeared in 25% (4/16) with no significant changes in voided volume. An improvement was observed in 66% (16/24) of patients with isolated voiding symptoms, in 51.2% (20/39) with associated storage symptoms and in 65% (17/26) of women with associated recurrent urinary infections. Conclusions: These results suggest that α-blocker may be an effective treatment option in women with voiding dysfunction due to functional BOO.
The Effect of Phosphodiesterase Isoenzyme 5 Inhibitors on Ureteropelvic Junction Obstruction in Rabbits
Authors: Wang Hong Gen , Yan Xing Yu , Cai You Mei , Wang Hong Geng , Chen Jun Yi
Published: October 2009

Objective: To determine the effects of phosphodiesterase isoenzyme 5 (PDE5) inhibitors upon ureteropelvic junction obstruction (UPJO) in rabbits. Materials and Methods: Twenty-four New Zealand rabbits were randomly divided into three groups: sham-operated animals (group A; right-sided operations comprised subgroup A1 and left-sided operations comprised subgroup A2), operated animals (group B) and operated animals that received vardenafil (group C). The right-sided junctions of group A (A1) were not exposed, and therefore made up the control group. The UPJO model was established by partial obstruction of the left junction. Intravenous pyelograms were performed before and after the operation. HE staining and microscopic examinations were performed to chart the pathological changes. Immunohistochemistry (streptavidin peroxidase) was used to test the expression of TGF-β1 and nNOS in the junctions. RT-PCR detected mRNA expression of TGF-β1. Results: We observed serious hydronephrosis in group B and moderate hydronephrosis in group C, but not in group A. We also observed large pathological changes in group B, but little change in group C and no change in subgroups A1 and A2. The level of TGF-β1 in group B was significantly higher than in groups A and C; group C expressed more TGF-β1 than group A. More nNOS was detected in group C than group B, although the two groups both expressed nNOS at lower levels than group A. Conclusions: More TGF-β1 and less nNOS are expressed when the junction is obstructed. PDE5 inhibitors may be able to regulate these 2 factors, or other factors that have not been discussed in this experiment, in order to halt the progression of UPJO.
The Use of a Ureteral Access Sheath for the Urethral Dilatation and Catheterization of Difficult Urethral Strictures
Authors: Athanasopoulos Anastasios , Liatsikos Evangelos N.
Published: October 2009

Background: Urethral catheterization can often be technically demanding due to the presence of obstructing segments within the urethra. Conventional methods attempting to restore urethral patency often are unsuccessful and traumatize the urethral lumen. Method: We present an innovative technique by using a hydrophilic guidewire and a hydrophilic ureteral access sheath for the ‘first-line’ catheterization and management of ‘difficult’ urethral strictures. We used this technique in 10 extremely difficult cases of strictures present in all the urethral segments. Result: The dilation was successful and atraumatic in all the cases due to the hydrophilic nature of the access sheath. In all patients, larger sizes of urethral catheters were gradually inserted within the next couple of days. Conclusion: We propose ‘access sheath’-assisted urethral dilatation as a safe and simple alternative for the ‘first-line’ catheterization and, possibly, management of ‘difficult’ urethral strictures.
Perivesical Inflammation and Necrosis due to Mitomycin C Instillation after Transurethral Resection of Bladder Tumor: We Must Be Vigilant!
Authors: Fazlıoglu Adem , Tandogdu Zafer , Kurtulus Fatih Osman , Parlakkilic Oguzhan , Cek Mete
Published: October 2009

The instillation of chemotherapeutic agents after transurethral resection of bladder tumor as an adjuvant treatment in bladder tumors is accepted as being safe if there is no associated bladder perforation. Mitomycin C (MMC) is a widely used agent for the adjuvant treatment of bladder tumors. A patient who developed MMC-induced bladder perforation and perivesical necrosis after an uneventful endoscopic tumor is presented. We suggest that the possible reason for such a complication could be deep resection at the anterior bladder wall. The treatment of the situation continued for an extended period, causing serious discomfort for the patient. Withholding immediate intravesical MMC instillation after resection of tumors on the anterior bladder wall ought to be considered in the early postoperative hours.
Multimodal Tumor Therapy in a 31-Year-Old Pregnant Woman with Wilms Tumor
Authors: Maurer T. , Zorn C. , Klein E. , Weirich G. , Beer A.J. , Gschwend J.E. , Zantl N.
Published: October 2009

Wilms tumor, or nephroblastoma, is the most common malignant tumor of the urinary tract in children, but is rarely found in adults. Here, we report the first case of a female patient with a Wilms tumor, diagnosed during pregnancy, who underwent radical nephrectomy and adjuvant chemotherapy before and after delivering a healthy child. Generally, treatment should follow the guidelines established for the pediatric setting.
Malignant Peripheral Nerve Sheath Tumor of the Renal Pelvis
Authors: Voznesensky Maria A. , Yamase Harold , Taylor, III John A.
Published: October 2009

Malignant peripheral nerve sheath tumors (MPNST) of the urothelial collecting system are rare. These tumors contain both malignant epithelial and sarcomatous components. Metastatic spread of MPNST is common, however, metastases to the kidney and perirenal tissues are rare. There are only 4 cases reported of MPNST of the kidney. We report a rare case of MPNST arising from the submucosa of the renal pelvis.