19 results on '"Katsaros, D"'
Search Results
2. A combined array-based comparative genomic hybridization and functional library screening approach identifies mir-30d as an oncomir in cancer.
- Author
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Li N, Kaur S, Greshock J, Lassus H, Zhong X, Wang Y, Leminen A, Shao Z, Hu X, Liang S, Katsaros D, Huang Q, Bützow R, Weber BL, Coukos G, and Zhang L
- Subjects
- Animals, Apoptosis genetics, Cell Proliferation, Chromosome Mapping, Female, Humans, In Situ Hybridization, Mice, Ovarian Neoplasms pathology, Polymorphism, Single Nucleotide, Real-Time Polymerase Chain Reaction, Comparative Genomic Hybridization, MicroRNAs genetics, Ovarian Neoplasms genetics
- Abstract
Oncomirs are microRNAs (miRNA) that acts as oncogenes or tumor suppressor genes. Efficient identification of oncomirs remains a challenge. Here we report a novel, clinically guided genetic screening approach for the identification of oncomirs, identifying mir-30d through this strategy. mir-30d regulates tumor cell proliferation, apoptosis, senescence, and migration. The chromosomal locus harboring mir-30d was amplified in more than 30% of multiple types of human solid tumors (n = 1,283). Importantly, higher levels of mir-30d expression were associated significantly with poor clinical outcomes in ovarian cancer patients (n = 330, P = 0.0016). Mechanistic investigations suggested that mir-30d regulates a large number of cancer-associated genes, including the apoptotic caspase CASP3. The guided genetic screening approach validated by this study offers a powerful tool to identify oncomirs that may have utility as biomarkers or targets for drug development., (©2011 AACR.)
- Published
- 2012
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3. Double-negative feedback loop between reprogramming factor LIN28 and microRNA let-7 regulates aldehyde dehydrogenase 1-positive cancer stem cells.
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Yang X, Lin X, Zhong X, Kaur S, Li N, Liang S, Lassus H, Wang L, Katsaros D, Montone K, Zhao X, Zhang Y, Bützow R, Coukos G, and Zhang L
- Subjects
- 3' Untranslated Regions genetics, Adult, Aged, Aldehyde Dehydrogenase genetics, Aldehyde Dehydrogenase 1 Family, Animals, Binding Sites genetics, Blotting, Western, Cell Line, Tumor, Cells, Cultured, DNA-Binding Proteins genetics, Feedback, Physiological, Female, Gene Expression Profiling, HeLa Cells, Humans, Immunohistochemistry, Isoenzymes genetics, Mice, Mice, Inbred C57BL, MicroRNAs genetics, Middle Aged, NIH 3T3 Cells, Neoplastic Stem Cells pathology, Oligonucleotide Array Sequence Analysis, Protein Binding, RNA Interference, RNA-Binding Proteins, Retinal Dehydrogenase, Aldehyde Dehydrogenase metabolism, DNA-Binding Proteins metabolism, Isoenzymes metabolism, MicroRNAs metabolism, Neoplastic Stem Cells metabolism
- Abstract
A relatively rare aldehyde dehydrogenase 1 (ALDH1)-positive "stem cell-like" subpopulation of tumor cells has the unique ability to initiate and perpetuate tumor growth; moreover, it is highly resistant to chemotherapy and significantly associated with poor clinical outcomes. The development of more effective therapies for cancer requires targeting of this cell population. Using cDNA microarray analysis, we identified that the expression of the Caenorhabditis elegans lin-28 homologue (LIN28) was positively correlated with the percentage of ALDH1+ tumor cells; this was further validated in an independent set of tissue arrays (n=197). Both loss-of-function and gain-of-function studies showed that LIN28 plays a critical role in the maintenance of ALDH1+ tumor cells. In addition, we found that there is a double-negative feedback loop between LIN28 and let-7 in tumor cells, and that let-7 negatively regulates ALDH1+ tumor cells. Finally, we report that a LIN28/let-7 loop modulates self-renewal and differentiation of mammary gland epithelial progenitor cells. Our data provide evidence that cancer stem cells may arise through a "reprogramming-like" mechanism. A rebalancing of the LIN28/let-7 regulatory loop could be a novel therapeutic strategy to target ALDH1+ cancer stem cells., (Copyright © 2010 AACR.)
- Published
- 2010
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4. MicroRNA microarray identifies Let-7i as a novel biomarker and therapeutic target in human epithelial ovarian cancer.
- Author
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Yang N, Kaur S, Volinia S, Greshock J, Lassus H, Hasegawa K, Liang S, Leminen A, Deng S, Smith L, Johnstone CN, Chen XM, Liu CG, Huang Q, Katsaros D, Calin GA, Weber BL, Bützow R, Croce CM, Coukos G, and Zhang L
- Subjects
- Adult, Aged, Aged, 80 and over, Biomarkers, Tumor biosynthesis, Cisplatin pharmacology, DNA, Neoplasm genetics, Drug Resistance, Neoplasm, Female, Gene Dosage, Humans, MicroRNAs biosynthesis, Middle Aged, Oligonucleotide Array Sequence Analysis, Ovarian Neoplasms metabolism, RNA, Messenger genetics, Young Adult, Biomarkers, Tumor genetics, MicroRNAs genetics, Ovarian Neoplasms drug therapy, Ovarian Neoplasms genetics
- Abstract
MicroRNAs (miRNA) are approximately 22-nucleotide noncoding RNAs that negatively regulate protein-coding gene expression in a sequence-specific manner via translational inhibition or mRNA degradation. Our recent studies showed that miRNAs exhibit genomic alterations at a high frequency and their expression is remarkably deregulated in ovarian cancer, strongly suggesting that miRNAs are involved in the initiation and progression of this disease. In the present study, we performed miRNA microarray to identify the miRNAs associated with chemotherapy response in ovarian cancer and found that let-7i expression was significantly reduced in chemotherapy-resistant patients (n = 69, P = 0.003). This result was further validated by stem-loop real-time reverse transcription-PCR (n = 62, P = 0.015). Both loss-of-function (by synthetic let-7i inhibitor) and gain-of-function (by retroviral overexpression of let-7i) studies showed that reduced let-7i expression significantly increased the resistance of ovarian and breast cancer cells to the chemotherapy drug, cis-platinum. Finally, using miRNA microarray, we found that decreased let-7i expression was significantly associated with the shorter progression-free survival of patients with late-stage ovarian cancer (n = 72, P = 0.042). This finding was further validated in the same sample set by stem-loop real-time reverse transcription-PCR (n = 62, P = 0.001) and in an independent sample set by in situ hybridization (n = 53, P = 0.049). Taken together, our results strongly suggest that let-7i might be used as a therapeutic target to modulate platinum-based chemotherapy and as a biomarker to predict chemotherapy response and survival in patients with ovarian cancer.
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- 2008
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5. Hypermethylation of let-7a-3 in epithelial ovarian cancer is associated with low insulin-like growth factor-II expression and favorable prognosis.
- Author
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Lu L, Katsaros D, de la Longrais IA, Sochirca O, and Yu H
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- Adult, Aged, Aged, 80 and over, Female, Humans, Insulin-Like Growth Factor Binding Protein 3, Insulin-Like Growth Factor Binding Proteins genetics, Middle Aged, Neoplasms, Glandular and Epithelial mortality, Ovarian Neoplasms mortality, Prognosis, RNA, Messenger analysis, DNA Methylation, Insulin-Like Growth Factor II genetics, MicroRNAs genetics, Neoplasms, Glandular and Epithelial genetics, Ovarian Neoplasms genetics
- Abstract
MicroRNAs (miRNA) are endogenous noncoding small RNAs that regulate the activity of mRNAs. Many miRNA genes, including let-7a-3, are located in CpG islands, suggesting possible epigenetic regulation of their expression. Promoter CpG island methylation of tumor suppressor genes is involved in cancer development and progression. Using real-time methylation-specific PCR and real-time reverse transcription-PCR, we analyzed DNA methylation in the let-7a-3 gene and miRNA expression of let-7a in 214 patients with epithelial ovarian cancer to assess the effect of let-7a-3 methylation on the expressions of let-7a as well as a possible target of let-7 regulation, insulin-like growth factor-II (IGF-II). The association of let-7a-3 methylation with patient survival outcomes was also evaluated. let-7a-3 methylation was detected in epithelial ovarian cancer, and the expression of let-7a was slightly affected by the methylation, but the effect was not substantial. The methylation of let-7a-3, however, was inversely correlated with IGF-II expression and positively with insulin-like growth factor binding protein-3 (IGFBP-3) expression. Patients with methylated let-7a-3 seemed to have reduced risk for death compared with those without, and the association was independent of patient age at surgery, tumor grade, disease stage, and IGF-II or IGFBP-3 expression. No association was found for let-7a-3 methylation and disease progression. These results suggest that the let-7a-3 gene is methylated and the methylation may affect IGF-II expression and the survival of ovarian cancer patients. Further investigation of the role of miRNAs and their regulation in cancer is warranted.
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- 2007
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6. Integrative genomic analysis of protein kinase C (PKC) family identifies PKCiota as a biomarker and potential oncogene in ovarian carcinoma.
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Zhang L, Huang J, Yang N, Liang S, Barchetti A, Giannakakis A, Cadungog MG, O'Brien-Jenkins A, Massobrio M, Roby KF, Katsaros D, Gimotty P, Butzow R, Weber BL, and Coukos G
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- Cell Growth Processes physiology, Cell Line, Tumor, Cell Transformation, Neoplastic genetics, Cell Transformation, Neoplastic metabolism, Cisplatin pharmacology, Cisplatin therapeutic use, Drug Resistance, Neoplasm, Female, Gene Dosage, Humans, Isoenzymes biosynthesis, Mutation, Ovarian Neoplasms drug therapy, Protein Kinase C biosynthesis, RNA, Messenger biosynthesis, RNA, Messenger genetics, Transcription, Genetic, Transfection, Up-Regulation, ras Proteins genetics, Biomarkers, Tumor genetics, Isoenzymes genetics, Ovarian Neoplasms enzymology, Ovarian Neoplasms genetics, Protein Kinase C genetics
- Abstract
The protein kinase C (PKC) family plays a key regulatory role in a wide range of cellular functions as well as in various cancer-associated signal transduction pathways. Here, we investigated the genomic alteration and gene expression of most known PKC family members in human ovarian cancer. The DNA copy number of PKC family genes was screened by a high-resolution array-based comparative genomic hybridization in 89 human ovarian cancer specimens. Five PKC genes exhibited significant DNA copy number gains, including PKCiota (43.8%), PKCbeta1 (37.1%), PKCgamma (27.6%), PKCzeta (22.5%), and PKCtheta (21.3%). None of the PKC genes exhibited copy number loss. The mRNA expression level of PKC genes was analyzed by microarray retrieval approach. Two of the amplified PKC genes, PKCiota and PKCtheta, were significantly up-regulated in ovarian cancer compared with normal ovary. Increased PKCiota expression correlated with tumor stage or grade, and PKCiota overexpression was seen mostly in ovarian carcinoma but not in other solid tumors. The above results were further validated by real-time reverse transcription-PCR with 54 ovarian cancer specimens and 24 cell lines; overexpression of PKCiota protein was also confirmed by tissue array and Western blot. Interestingly, overexpressed PKCiota did not affect ovarian cancer cell proliferation or apoptosis in vitro. However, decreased PKCiota expression significantly reduced anchorage-independent growth of ovarian cancer cells, whereas overexpression of PKCiota contributed to murine ovarian surface epithelium transformation in cooperation with mutant Ras. We propose that PKCiota may serve as an oncogene and a biomarker of aggressive disease in human ovarian cancer.
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- 2006
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7. Transcriptional coactivator Drosophila eyes absent homologue 2 is up-regulated in epithelial ovarian cancer and promotes tumor growth.
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Zhang L, Yang N, Huang J, Buckanovich RJ, Liang S, Barchetti A, Vezzani C, O'Brien-Jenkins A, Wang J, Ward MR, Courreges MC, Fracchioli S, Medina A, Katsaros D, Weber BL, and Coukos G
- Subjects
- Animals, Cell Growth Processes genetics, DNA, Neoplasm genetics, Disease Progression, Female, Gene Amplification, Gene Expression Regulation, Neoplastic, Humans, Intracellular Signaling Peptides and Proteins, Mice, Mice, Inbred BALB C, Nuclear Proteins, Ovarian Neoplasms metabolism, Ovarian Neoplasms pathology, Prognosis, Protein Tyrosine Phosphatases, Trans-Activators biosynthesis, Transcriptional Activation genetics, Up-Regulation, Ovarian Neoplasms genetics, Trans-Activators genetics
- Abstract
Epithelial ovarian cancer is the most frequent cause of gynecologic malignancy-related mortality in women. To identify genes up-regulated in ovarian cancer, PCR-select cDNA subtraction was done and Drosophila Eyes Absent Homologue 2 (EYA2) was isolated as a promising candidate. The transcriptional coactivator eya controls essential cellular functions during organogenesis of Drosophila. EYA2 mRNA was found to be up-regulated in ovarian cancer by real-time reverse transcription-PCR, whereas its protein product was detected in 93.6% of ovarian cancer specimens by immunohistochemistry (n = 140). EYA2 was amplified in 14.8% of ovarian carcinomas, as detected by array-based comparative genomic hybridization (n = 88). Most importantly, EYA2 overexpression was significantly associated with short overall survival in advanced ovarian cancer (n = 99, P = 0.0361). EYA2 was found to function as transcriptional activator in ovarian cancer cells by Gal4 assay and to promote tumor growth in vivo in xenograft models. Therefore, this study suggests an important role of EYA2 in ovarian cancer and its potential application as a therapeutic target.
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- 2005
8. Involvement of estrogen receptor beta in ovarian carcinogenesis.
- Author
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Bardin A, Hoffmann P, Boulle N, Katsaros D, Vignon F, Pujol P, and Lazennec G
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- Adenoviridae genetics, Apoptosis physiology, Cell Division physiology, Cell Line, Tumor, Cell Movement physiology, Estrogen Receptor alpha, Estrogen Receptor beta, Female, Humans, Middle Aged, Ovarian Neoplasms genetics, Ovarian Neoplasms pathology, RNA, Messenger biosynthesis, RNA, Messenger genetics, Receptors, Estrogen genetics, Reverse Transcriptase Polymerase Chain Reaction, Ovarian Neoplasms metabolism, Receptors, Estrogen biosynthesis, Receptors, Estrogen physiology
- Abstract
Knockout and expression studies suggest that estrogen receptor beta (ERbeta) plays a prominent role in ovarian function and pathology. Moreover, ovarian cancers are characterized by high morbidity and low responsiveness to anti-estrogens. Here we demonstrate, using quantitative PCR to measure ERalpha and ERbeta levels in 58 ovarian cancer patients, that ERbeta expression decreased in cysts and ovarian carcinomas as compared with normal ovaries and that this decrease is attributable only to a selective loss in ERbeta expression during cancer progression. To address the question of a possible involvement of ERbeta in ovarian cancers, we restored ERalpha and ERbeta expression in two human ovarian cancer cell lines PEO14 (ERalpha-negative) and BG1 (ERalpha-positive) using adenoviral delivery. ERalpha, but not ERbeta, could induce progesterone receptor and fibulin-1C. Moreover, ERalpha and ERbeta had opposite actions on cyclin D1 gene regulation, because ERbeta down-regulated cyclin D1 gene expression, whereas ERalpha increased cyclin D1 levels. Interestingly, ERbeta expression strongly inhibited PEO14 and BG1 cell proliferation and cell motility in a ligand-independent manner, whereas ERalpha had no marked effect. Induction of apoptosis by ERbeta also contributed to the decreased proliferation of ovarian cancer cells, as shown by Annexin V staining. This study shows that ERbeta is an important regulator of proliferation and motility of ovarian cancer and provides the first evidence for a proapoptotic role of ERbeta. The loss of ERbeta expression may thus be an important event leading to the development of ovarian cancer.
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- 2004
- Full Text
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9. Ovarian carcinoma expresses the NKG2D ligand Letal and promotes the survival and expansion of CD28- antitumor T cells.
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Conejo-Garcia JR, Benencia F, Courreges MC, Gimotty PA, Khang E, Buckanovich RJ, Frauwirth KA, Zhang L, Katsaros D, Thompson CB, Levine B, and Coukos G
- Subjects
- Adenocarcinoma, Mucinous immunology, Adenocarcinoma, Mucinous pathology, Animals, Apoptosis, Carcinoma, Endometrioid immunology, Carcinoma, Endometrioid pathology, Cell Survival, Cystadenocarcinoma, Serous immunology, Cystadenocarcinoma, Serous pathology, Cytotoxicity, Immunologic, Female, Humans, Ligands, Lymphocyte Activation, Mice, Mice, Inbred C57BL, NK Cell Lectin-Like Receptor Subfamily K, Ovarian Neoplasms pathology, Receptors, Natural Killer Cell, CD28 Antigens metabolism, CD8-Positive T-Lymphocytes immunology, Lymphocytes, Tumor-Infiltrating immunology, Ovarian Neoplasms immunology, Receptors, Immunologic metabolism
- Abstract
The role of the NKG2D immunoreceptor and its ligands in antitumor immune response is incompletely understood. Here, we report that effector immune cells infiltrating ovarian carcinoma are mostly CD8+ lymphocytes lacking CD28 but expressing the NKG2D costimulatory receptor. Human ovarian carcinoma expresses the novel NKG2D ligand lymphocyte effector cell toxicity-activating ligand (Letal). Letal was found to be an independent prognosticator of improved survival in advanced ovarian cancer. Higher levels of tumor-derived Letal were associated with stronger lymphocyte infiltration. Letal exerted marked costimulatory effects and induced type-1 polarization in CD8+CD28- tumor-infiltrating lymphocytes ex vivo. Letal engagement increased the expression of the glucose transporter Glut-1, enhanced glucose up-take, and protected CD8+ lymphocytes from cisplatin-induced killing. Letal also down-regulated the expression of Fas in CD8+ cells and rendered them resistant to Fas ligand-induced apoptosis. Our results indicate that Letal promotes tumor immune surveillance by promoting the survival and intratumoral expansion of antitumor cytotoxic lymphocytes. We propose that Letal could be used for the ex vivo expansion of apoptosis-resistant tumor-reactive cytotoxic lymphocytes for adoptive transfer.
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- 2004
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10. Human kallikrein 14: a new potential biomarker for ovarian and breast cancer.
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Borgoño CA, Grass L, Soosaipillai A, Yousef GM, Petraki CD, Howarth DH, Fracchioli S, Katsaros D, and Diamandis EP
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- Amino Acid Sequence, Animals, Antibodies immunology, Biomarkers, Tumor blood, Biomarkers, Tumor genetics, Breast Neoplasms blood, Cell Line, Tumor, Cytosol metabolism, Enzyme-Linked Immunosorbent Assay, Female, Humans, Kallikreins blood, Kallikreins genetics, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Ovarian Neoplasms blood, Recombinant Proteins biosynthesis, Recombinant Proteins genetics, Sensitivity and Specificity, Biomarkers, Tumor metabolism, Breast Neoplasms metabolism, Kallikreins metabolism, Neoplasms, Hormone-Dependent metabolism, Ovarian Neoplasms metabolism
- Abstract
Human kallikrein gene 14 (KLK14) is a recently discovered member of the tissue kallikrein family of secreted serine proteases, which includes hK3/prostate-specific antigen, the best cancer biomarker to date. Given that KLK14 is hormonally regulated, differentially expressed in endocrine-related cancers, and a prognostic marker for breast and ovarian cancer at the mRNA level, we hypothesize that its encoded protein, hK14, like hK3/prostate-specific antigen, may constitute a new biomarker for endocrine-related malignancies. The objective of this study was to generate immunological reagents for hK14, to develop an ELISA and immunohistochemical techniques to study its expression in normal and cancerous tissues and biological fluids. Recombinant hK14 was produced in Pichia pastoris, purified by affinity chromatography, and injected into mice and rabbits for polyclonal antibody generation. Using the mouse and rabbit antisera, a sandwich-type immunofluorometric ELISA and immunohistochemical methodologies were developed for hK14. The ELISA was sensitive (detection limit of 0.1 micro g/liter), specific for hK14, linear from 0 to 20 micro g/liter with between-run and within-run coefficients of variation of <10%. hK14 was quantified in human tissue extracts and biological fluids. Highest levels were observed in the breast, skin, prostate, seminal plasma, and amniotic fluid, with almost undetectable levels in normal serum. hK14 concentration was higher in 40% of ovarian cancer tissues compared with normal ovarian tissues. Serum hK14 levels were elevated in a proportion of patients with ovarian (65%) and breast (40%) cancers. Immunohistochemical analyses indicated strong cytoplasmic staining of hK14 by the epithelial cells of normal and malignant skin, ovary, breast, and testis. In conclusion, we report the first ELISA and immunohistochemical assays for hK14 and describe its distribution in tissues and biological fluids. Our preliminary data indicate that hK14 is a potential biomarker for breast and ovarian cancers.
- Published
- 2003
11. The oncogene phosphatidylinositol 3'-kinase catalytic subunit alpha promotes angiogenesis via vascular endothelial growth factor in ovarian carcinoma.
- Author
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Zhang L, Yang N, Katsaros D, Huang W, Park JW, Fracchioli S, Vezzani C, Rigault de la Longrais IA, Yao W, Rubin SC, and Coukos G
- Subjects
- Apoptosis physiology, Catalytic Domain, Cell Division physiology, Chromones pharmacology, Endothelial Growth Factors genetics, Enzyme Inhibitors pharmacology, Female, Gene Expression Regulation, Enzymologic, Gene Expression Regulation, Neoplastic, Humans, Hypoxia-Inducible Factor 1, alpha Subunit, Intercellular Signaling Peptides and Proteins genetics, Lymphokines genetics, Morpholines pharmacology, Neovascularization, Pathologic enzymology, Ovarian Neoplasms enzymology, Ovarian Neoplasms genetics, Phosphatidylinositol 3-Kinases biosynthesis, Phosphatidylinositol 3-Kinases metabolism, Phosphoinositide-3 Kinase Inhibitors, RNA, Messenger biosynthesis, RNA, Messenger genetics, Transcription Factors metabolism, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors, Endothelial Growth Factors biosynthesis, Intercellular Signaling Peptides and Proteins biosynthesis, Lymphokines biosynthesis, Neovascularization, Pathologic genetics, Ovarian Neoplasms blood supply, Phosphatidylinositol 3-Kinases genetics
- Abstract
The gene of phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA) has been implicated as an oncogene in ovarian cancer [L. Shayesteh et al., Nat. Genet., 21: 99-102, 1999]. In this study, we examined the expression of PIK3CA mRNA and its p110alpha protein product in human ovarian carcinoma and investigated its role in regulating angiogenesis via vascular endothelial growth factor (VEGF). PIK3CA mRNA was detected in 66.6% of stage I and 93.9% of advanced stage ovarian cancer specimens and in all 17 ovarian cancer cell lines. PIK3CA mRNA levels were significantly higher in invasive carcinomas compared with benign and low malignant potential neoplasms (P = 0.007), but no significant difference was seen between early and advanced stage carcinomas (P = 0.812). Strong expression of immunoreactive p110alpha was detected in tumor cells and/or stroma endothelium. PIK3CA expression in vivo positively correlated, both at the mRNA and the protein level, with the expression of VEGF as well as with the extent of microvascular development. Furthermore, PIK3CA mRNA overexpression positively correlated with increased proliferation and decreased apoptosis of tumor cells in vivo. In vitro, PIK3CA expression positively correlated with the expression of VEGF in ovarian cancer cells, whereas the phosphatidylinositol 3'-kinase inhibitor Ly294002 reduced both the constitutive and inducible expression of hypoxia-inducible factor-1alpha at the mRNA and protein levels and abrogated VEGF up-regulation by glucose starvation. Furthermore, Ly294002 suppressed cell proliferation and, at higher doses, induced marked apoptosis in ovarian cancer cells. Collectively, these data strongly indicate that PIK3CA supports ovarian cancer growth through multiple and independent pathways affecting cell proliferation, apoptosis and angiogenesis, and plays an important role in ovarian cancer progression.
- Published
- 2003
12. Tumor-derived vascular endothelial growth factor up-regulates angiopoietin-2 in host endothelium and destabilizes host vasculature, supporting angiogenesis in ovarian cancer.
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Zhang L, Yang N, Park JW, Katsaros D, Fracchioli S, Cao G, O'Brien-Jenkins A, Randall TC, Rubin SC, and Coukos G
- Subjects
- Angiogenesis Inducing Agents genetics, Angiopoietin-1, Angiopoietin-2, Animals, Carcinoma blood supply, Carcinoma metabolism, Carcinoma pathology, Cell Line, Transformed drug effects, Cell Line, Transformed metabolism, Dactinomycin pharmacology, Endothelial Growth Factors pharmacology, Endothelium, Vascular metabolism, Female, Genes, Reporter, Humans, Intercellular Signaling Peptides and Proteins pharmacology, Lymphokines pharmacology, Membrane Glycoproteins biosynthesis, Membrane Glycoproteins genetics, Mice, Mice, Inbred C57BL, Models, Biological, Neoplasm Proteins biosynthesis, Neoplasm Proteins genetics, Neoplasm Proteins pharmacology, Nucleic Acid Synthesis Inhibitors pharmacology, Ovarian Neoplasms blood supply, Ovarian Neoplasms metabolism, Ovarian Neoplasms pathology, Pericytes pathology, RNA, Messenger biosynthesis, RNA, Neoplasm biosynthesis, Receptor Protein-Tyrosine Kinases biosynthesis, Receptor Protein-Tyrosine Kinases genetics, Receptor, TIE-2, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins physiology, Stromal Cells drug effects, Stromal Cells metabolism, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors, Angiogenesis Inducing Agents biosynthesis, Carcinoma physiopathology, Endothelial Growth Factors physiology, Endothelium, Vascular drug effects, Gene Expression Regulation, Neoplastic, Intercellular Signaling Peptides and Proteins physiology, Lymphokines physiology, Neoplasm Proteins physiology, Neovascularization, Pathologic physiopathology, Ovarian Neoplasms physiopathology, Paracrine Communication
- Abstract
Vascular remodeling in host tissues surrounding growing tumors is implicated in the successful development of tumor neovasculature. Cooperation between vascular endothelial growth factor (VEGF) and angiopoietins (Angs) is considered to be critical in this context. However, the mechanisms regulating the coordinated expression of these molecules remain, to date, elusive. In this study, we used a murine ovarian cancer angiogenesis model induced by overexpression of VEGF, as well as 52 human ovarian cancer specimens and 36 established cancer cell lines to characterize the expression and regulation of Ang-2 in the context of tumor angiogenesis. Using a combination of immunohistochemistry, laser capture microdissection and real-time quantitative reverse transcription-PCR, we showed that tumor-derived VEGF significantly up-regulated the expression of Ang-2 in host stroma endothelial cells, resulting in markedly increased Ang-2/Tie-2 mRNA copy number ratio in vivo. In vitro experiments showed that VEGF directly up-regulated Ang-2, which is mediated via VEGF receptor-2/flk-1/KDR pathway, in cultured endothelial cells through transcriptional activation rather than the enhanced mRNA stability. In human ovarian cancer, Ang-2 was primarily expressed in stroma endothelial cells and detectable in tumor cells of only 12% tumor specimens; however, it was not detected in the majority of established ovarian cancer cell lines. In addition, a significant correlation was observed between VEGF and Ang-2 mRNA expression (P < 0.01) but not between VEGF and Ang-1 or Tie-2 in human ovarian cancer specimens. In the mouse ovarian cancer model, up-regulation of Ang-2 in host stroma endothelial cells was significantly associated with pericyte loss and instability of the host vasculature surrounding the tumor. Our study suggests a novel mechanism by which tumor-derived VEGF interacts with Angs/Tie-2 system in host stroma endothelial cells and induces in a paracrine manner the remodeling of host vasculature to support angiogenesis during tumor growth.
- Published
- 2003
13. Parallel overexpression of seven kallikrein genes in ovarian cancer.
- Author
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Yousef GM, Polymeris ME, Yacoub GM, Scorilas A, Soosaipillai A, Popalis C, Fracchioli S, Katsaros D, and Diamandis EP
- Subjects
- Computational Biology methods, Female, Gene Expression Regulation, Neoplastic, Humans, Kallikreins biosynthesis, Ovarian Neoplasms metabolism, Reproducibility of Results, Tumor Cells, Cultured, Up-Regulation, Kallikreins genetics, Ovarian Neoplasms genetics
- Abstract
Recent evidence suggests that many members of the human kallikrein (KLK) gene family are differentially regulated in ovarian cancer and have potential as diagnostic and/or prognostic markers. We used the serial analysis of gene expression and expressed sequence tag databases of the Cancer Genome Anatomy Project to perform in silico analyses of the expression pattern of the 15 human KLK genes in normal and cancerous ovarian tissues and cell lines. We found that seven KLK genes (KLK5, KLK6, KLK7, KLK8, KLK10, KLK11, and KLK14) are up-regulated in ovarian cancer. Probing 2 normal and 10 ovarian cancer serial analysis of gene expression libraries with gene-specific tags for each KLK indicated that whereas no expression was detected in any normal libraries (with the exception of KLK10 and KLK11), these KLKs were found to be expressed with moderate densities (103-408 tags per million) in 40-60% of the ovarian cancer libraries analyzed. These data were verified by screening the expressed sequence tag databases, where 78 of 79 mRNA clones isolated for these genes were from ovarian cancer libraries. X-profiler comparison of the pools of normal and cancerous ovaries identified a significant difference in expression levels for six of the seven KLKs. We experimentally verified the overexpression of six KLK proteins in cancer versus normal or benign tissues with highly sensitive and specific immunofluorometric assays. A statistically significant stepwise increase in protein levels was found among normal, benign, and cancerous ovarian tissues. The expression of five KLKs showed a strong degree of correlation at the protein level, suggesting the existence of a common mechanism or pathway that controls the expression of this group of adjacent genes during ovarian cancer progression.
- Published
- 2003
14. The serum concentration of human kallikrein 10 represents a novel biomarker for ovarian cancer diagnosis and prognosis.
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Luo LY, Katsaros D, Scorilas A, Fracchioli S, Bellino R, van Gramberen M, de Bruijn H, Henrik A, Stenman UH, Massobrio M, van der Zee AG, Vergote I, and Diamandis EP
- Subjects
- Adult, Aged, CA-125 Antigen blood, Female, Fluoroimmunoassay, Humans, Middle Aged, Neoplasm Staging, Ovarian Neoplasms pathology, Ovarian Neoplasms surgery, Prognosis, Sensitivity and Specificity, Biomarkers, Tumor blood, Kallikreins blood, Ovarian Neoplasms blood
- Abstract
Human kallikrein 10 (hK10) is a secreted serine protease that is highly expressed in ovarian tissue. We hypothesized that hK10 might represent a novel serological marker for ovarian cancer. We quantified by immunoassay, hK10 in sera from 97 normal women (controls), 141 patients with benign gynecologic diseases, and 146 patients with ovarian cancer. We then examined the diagnostic and prognostic value of this measurement in ovarian cancer. We found that normal serum hK10 ranged from 50 to 1040 ng/liter (mean = 439 ng/liter). hK10 concentration is significantly elevated in serum of presurgical ovarian cancer patients (range: 106-11,746 ng/liter; mean = 1067 ng/liter) but not in serum of patients with benign gynecologic diseases (range: 120-1200 ng/liter; mean = 447 ng/liter). When a cutoff of 700 ng/liter was selected (diagnostic specificity = 90%), the diagnostic sensitivity for ovarian cancer is 54%. About 35% of CA125-negative ovarian cancer patients (CA125 < 23 kU/liter) were hK10 positive at 90% specificity. In patients with stage I/II ovarian cancer, use of these two markers in combination results in a 21% increase in sensitivity, at 90% specificity, compared with CA125 alone. High serum hK10 was strongly associated with serous epithelial type, late-stage, advanced grade, large residual tumor (>1 cm), suboptimal debulking, and no response to chemotherapy (all Ps < 0.001). In univariate Cox survival analysis, high serum hK10 is associated with increased risk for relapse and death (hazard ratio = 2.59 and 3.15, respectively, P = 0.003). This prognostic value remains significant for overall survival in the multivariate analysis. Kaplan-Meier survival curves demonstrated similar findings. Serum hK10 represents a novel biomarker for ovarian cancer. We conclude that preoperative serum hK10 concentration is a strong and independent unfavorable prognostic marker for ovarian cancer.
- Published
- 2003
15. Quantitative expression of the human kallikrein gene 9 (KLK9) in ovarian cancer: a new independent and favorable prognostic marker.
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Yousef GM, Kyriakopoulou LG, Scorilas A, Fracchioli S, Ghiringhello B, Zarghooni M, Chang A, Diamandis M, Giardina G, Hartwick WJ, Richiardi G, Massobrio M, Diamandis EP, and Katsaros D
- Subjects
- Adult, Aged, Aged, 80 and over, Analysis of Variance, Biomarkers, Tumor genetics, Estrogens physiology, Female, Gene Expression Regulation, Neoplastic, Humans, Immunohistochemistry, Kallikreins genetics, Middle Aged, Neoplasm Staging, Ovarian Neoplasms genetics, Ovarian Neoplasms pathology, Progestins physiology, Prognosis, Survival Rate, Up-Regulation, Biomarkers, Tumor biosynthesis, Kallikreins biosynthesis, Neoplasm Proteins, Ovarian Neoplasms metabolism
- Abstract
Many members of the human kallikrein gene family were found to be differentially expressed in various malignancies and some are useful cancer diagnostic/prognostic markers. KLK9 is a newly discovered human kallikrein gene that is expressed in several tissues including thymus, testis, spinal cord, salivary gland, ovary, and skin. Like other kallikreins, the KLK9 gene was found to be regulated by steroid hormones in cancer cell lines. Our purpose is to examine whether quantitative analysis of KLK9 expression has prognostic value in ovarian cancer. We studied the expression of KLK9 by quantitative reverse transcription-PCR in 168 consecutive ovarian tumors of different stages, grades, and histological types, and correlated the expression with clinicopathological parameters, response to chemotherapy, and patients' survival. We found that KLK9 expression was significantly higher in patients with early disease stages (I or II; P = 0.044) and in patients with optimal debulking (P = 0.019). Kaplan-Meier survival curves demonstrated that patients with KLK9-positive tumors have substantially longer progression-free and overall survival (P < 0.001 and P = 0.016, respectively). When the Cox proportional hazard regression analysis was applied to subgroups of patients, KLK9 expression was found to be a significant predictor of progression-free survival in the subgroup of patients with low-grade tumors [hazard ratio (HR), 0.13; P = 0.0015], early stage (HR, 0.099; P = 0.031); and those with optimal debulking (HR, 0.26; P = 0.012). After adjusting for other known prognostic variables, KLK9 retained its independent prognostic value in all of these subgroups of patients. A negative correlation was found between the expression levels of CA125 and KLK9 (rs, 0.350; P = 0.002). Our results indicate that KLK9 is under steroid hormone regulation in ovarian and breast cancer cell lines. Immmunohistochemically, human kallikrein protein (hK9) was localized in the cytoplasm, but not in the nuclei, of the epithelial cells of ovarian cancer tissues. We conclude that KLK9 is a potential new independent favorable prognostic marker for early stage, low-grade, optimally debulked ovarian cancer patients.
- Published
- 2001
16. Cloning of a new member of the human kallikrein gene family, KLK14, which is down-regulated in different malignancies.
- Author
-
Yousef GM, Magklara A, Chang A, Jung K, Katsaros D, and Diamandis EP
- Subjects
- Amino Acid Sequence, Base Sequence, Chromosome Mapping, Chromosomes, Human, Pair 19, Cloning, Molecular, Down-Regulation, Female, Gene Expression Regulation, Neoplastic, Humans, Kallikreins biosynthesis, Male, Molecular Sequence Data, Multigene Family genetics, Neoplasms metabolism, Phylogeny, Sequence Homology, Amino Acid, Kallikreins genetics, Neoplasms genetics
- Abstract
Kallikreins (KLKs) belong to the serine protease family of proteolytic enzymes. Human pancreatic/renal KLK (KLK1) encodes for an enzyme that is involved in posttranslational processing of polypeptide precursors. The function of the other members of this gene family is currently unknown, but growing evidence suggests that many KLKs are implicated in carcinogenesis. By using the positional candidate approach, we were able to identify a new human KLK-like gene, KLK14 (also known as KLK-L6). This new gene maps to chromosome 19q13.3-q13.4 and is formed of seven exons (two untranslated and five coding exons) and six intervening introns. KLK14 was defined as a KLK gene based on structural and mapping criteria, in relation to other known KLK genes. KLK14 is expressed in a variety of tissues, but the highest levels of KLK14 are found in the central nervous system, including brain, cerebellum, and spinal cord. Our preliminary results show that KLK14 is down-regulated, at the mRNA level, in breast, testicular, prostatic, and ovarian cancer.
- Published
- 2001
17. Prostate-specific antigen is a new favorable prognostic indicator for women with breast cancer.
- Author
-
Yu H, Giai M, Diamandis EP, Katsaros D, Sutherland DJ, Levesque MA, Roagna R, Ponzone R, and Sismondi P
- Subjects
- Adult, Aged, Aged, 80 and over, Breast Neoplasms mortality, Breast Neoplasms pathology, Cytosol chemistry, Disease-Free Survival, Female, Humans, Middle Aged, Multivariate Analysis, Receptors, Estrogen analysis, Survival Analysis, Biomarkers, Tumor analysis, Breast Neoplasms chemistry, Prostate-Specific Antigen analysis
- Abstract
Prostate-specific antigen (PSA) is thought to be produced exclusively by prostatic epithelial cells and is currently used as a tumor marker of prostatic adenocarcinoma. We recently found that 30% of breast cancers contain PSA immunoreactivity (IR-PSA). To examine the prognostic value of PSA in female breast cancer, we measured IR-PSA in tumor cytosols of 174 breast cancer patients and classified the breast cancers as either PSA positive or PSA negative based on an IR-PSA cutoff level of 0.03 ng/mg. IR-PSA was present in 27% of the patients. IR-PSA presence was associated with early disease stage, small tumors, and estrogen receptor-positive tumors. We used the Cox proportional hazards regression model to analyze survival of patients in association with PSA status and found that patients with IR-PSA-positive tumors had a reduced risk for relapse and death in univariate analysis (P = 0.02 and 0.06, respectively) and a reduced risk for relapse in multivariate analysis (P = 0.03). Further analysis indicated that the effect of IR-PSA on relapse-free survival was evident in node-positive or estrogen receptor-negative patients. Our study suggests that IR-PSA is an independent favorable prognostic marker for breast cancer and may be used to identify a subgroup of estrogen receptor-negative and/or node-positive patients who have good prognoses.
- Published
- 1995
18. Synergistic inhibition of growth of breast and colon human cancer cell lines by site-selective cyclic AMP analogues.
- Author
-
Tagliaferri P, Katsaros D, Clair T, Ally S, Tortora G, Neckers L, Rubalcava B, Parandoosh Z, Chang YA, and Revankar GR
- Subjects
- Chromatography, High Pressure Liquid, Cyclic AMP pharmacology, Drug Synergism, Female, Humans, Molecular Weight, Protein Kinases analysis, Proto-Oncogene Proteins analysis, Proto-Oncogene Proteins p21(ras), Receptors, Cyclic AMP analysis, Receptors, Cyclic AMP drug effects, Tumor Cells, Cultured drug effects, Breast Neoplasms pathology, Colonic Neoplasms pathology, Cyclic AMP analogs & derivatives
- Abstract
Our past studies on the mechanism of cyclic AMP (cAMP)-mediated control of tumor growth, using the experimental rat mammary tumor models as well as human breast cancer cell lines, indicated that the action of cAMP is mediated by the RII cAMP receptor protein, the regulatory subunit of cAMP-dependent protein kinase type II (Y. S. Cho-Chung, J. Cyclic Nucleotide Res., 6: 163, 1980). We now shown that the site-selective cAMP analogues, which are manyfold more active in binding to the cAMP receptor protein than previously studied analogues, demonstrate a potent growth inhibition of seven breast and three colon human cancer cell lines. The cAMP receptor protein has two different cAMP binding sites, and cAMP analogues that selectively bind to either one of the two binding sites are known as either site 1 selective (C-8 analogues) or site 2 selective (C-6 analogues). Nineteen site-selective analogues, C-6 and C-8 monosubstituted and C-6,-8 disubstituted, were tested for their growth regulatory effect. The majority of these analogues demonstrated an appreciable growth inhibition, with no sign of toxicity in all 10 cancer lines at micromolar concentrations. The three most potent inhibitors were 8-Cl-, N6-benzyl-, and N6-phenyl-8-thio-p-chlorophenyl-cAMP, demonstrating 50% growth inhibition at 5-25 microM concentrations (IC50). Furthermore, N6-analogues, in combination with halogen or thio derivatives of C-8 analogues, demonstrated synergistic enhancement of growth inhibition. The growth inhibition paralleled a change in cell morphology, an augmentation of the RII cAMP receptor protein, and a reduction in p21 ras protein. The growth inhibition by 8-Cl-cAMP was not due to its metabolite, 8-Cl-adenosine, since: (a) the growth inhibition by 8-Cl-cAMP was released upon cessation of treatment, whereas that by 8-Cl-adenosine was not released; (b) 8-Cl-cAMP treatment did not affect cell cycle progression, whereas 8-Cl-adenosine brought about G1 synchronization; (c) 8-Cl-cAMP treatment caused reduction of p21 ras protein, whereas 8-Cl-adenosine did not affect p21 levels; and (d) 8-Cl-adenosine was not detected in either cell extracts or medium from the cells treated with 8-Cl-cAMP for 48-72 h. Site-selective cAMP analogues thus provide a new physiological means to control the growth of breast and colon human cancer cells.
- Published
- 1988
19. Inhibition of growth and modulation of gene expression in human lung carcinoma in athymic mice by site-selective 8-Cl-cyclic adenosine monophosphate.
- Author
-
Ally S, Clair T, Katsaros D, Tortora G, Yokozaki H, Finch RA, Avery TL, and Cho-Chung YS
- Subjects
- 8-Bromo Cyclic Adenosine Monophosphate pharmacology, 8-Bromo Cyclic Adenosine Monophosphate therapeutic use, Animals, Blotting, Northern, Blotting, Western, Carcinoma drug therapy, Carcinoma metabolism, Cell Nucleus metabolism, Genes, ras, Humans, Isoenzymes metabolism, Lung Neoplasms drug therapy, Lung Neoplasms metabolism, Mice, Mice, Nude, Neoplasm Transplantation, Oncogenes, RNA, Messenger metabolism, RNA, Neoplasm metabolism, Carcinoma genetics, Cell Division drug effects, Gene Expression Regulation, Neoplastic drug effects, Lung Neoplasms genetics, Protein Kinases metabolism, Receptors, Cyclic AMP metabolism
- Abstract
Site-selective cyclic AMP (cAMP) analogues inhibit growth and induce changes in morphology in a spectrum of human cancer cell lines (D. Katsaros et al., FEBS Lett., 223:97, 1987). The cellular events underlying such effects of cAMP analogues include differential regulation of type I versus type II cAMP-dependent protein kinase isozymes (S. Ally et al., Proc. Natl. Acad. Sci. USA, 85: 6319, 1988). Infusion (i.p.) of 8-Cl-cAMP, the most potent site-selective cAMP analogue, for 7 days produced regression of LX-1 lung carcinoma in athymic mice in a dose-dependent manner. The tumor regression correlated with the changing levels of cAMP receptor proteins, RI alpha and RII beta, the regulatory subunits of cAMP-dependent protein kinase type I and type II, respectively. By photoaffinity labeling with 8-N3-[32P]cAMP and immunoblotting with a monospecific anti-RII antibody, RI alpha (Mr 49,000) and RII beta (Mr 51,000) were identified in the untreated control tumors. 8-Cl-cAMP treatment induced a rapid increase of both RI alpha and RII beta in tumor cytosols and translocation (within 1 h) of only RII beta from the cytosol to the nucleus. RII beta in both cytosols and nuclei remained elevated during 8-Cl-cAMP treatment, whereas RI alpha in the cytosols gradually decreased with time of treatment after its initial transient increase. Northern blot analyses demonstrated that the RII beta mRNA level increased within 6 h of 8-Cl-cAMP treatment and remained elevated during treatment, whereas the RI alpha mRNA level decreased to below that of the untreated control tumor level after its transient increase during 1-6 h of treatment. 8-Cl-cAMP treatment also caused a sharp decrease in both N-ras and c-myc mRNA levels. These results suggest that the fundamental basis for the antineoplastic activity of 8-Cl-cAMP may reside in the restoration of normal gene regulation in neoplasms in which cAMP receptor proteins play a role.
- Published
- 1989
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