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Ylermi Soini - One of the best experts on this subject based on the ideXlab platform.
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divergence of tight and adherens junction factors in alveolar epithelium in pulmonary fibrosis
Human Pathology, 2013Co-Authors: Elisa Lappiblanco, Siri Lehtonen, Heta Merikallio, Ylermi Soini, Raija Sormunen, Riitta KaarteenahoAbstract:Summary It has been proposed that an epithelial injury may be one of the multiple primary events in the pathogenesis of idiopathic pulmonary fibrosis (IPF). The aim of this study was to characterize the tight junction and adherens junction proteins in normal human lung, IPF, cryptogenic organizing pneumonia, and asbestosis. We determined the immunohistochemical cell-specific expression of tight junction proteins Claudin-1, Claudin-2, Claudin-3, Claudin-4, Claudin-5, and Claudin-7, as well as 3 adherens junction proteins, E-cadherin, N-cadherin, and β -catenin. We further analyzed the expression of Claudin-1, Claudin-3, and Claudin-4 and E-cadherin, N-cadherin, and β -catenin at the transcriptional level by quantitative real-time reverse transcriptase polymerase chain reaction. The expression levels of both tight junction and adherens junction proteins were elevated in regenerative alveolar epithelium in pulmonary fibrosis as compared with the expression of these proteins in normal alveolar epithelium. In particular, the expression levels of Claudins-1 and Claudin-3 were clearly elevated in all diseases. Furthermore, the amounts of adherens junction proteins messenger RNAs (mRNAs) were also all increased in pulmonary fibroses in comparison with healthy controls, with N-cadherin showing the greatest increase in mRNA levels in all diseases. However, the amounts of Claudin-1, Claudin-3, and Claudin-4 mRNAs in fibrotic lung were similar to or even lower than those measured in the healthy controls. It is possible that the diminished capacity to produce Claudin mRNAs may be one explanation for poor repair capacity of alveolar epithelial cells in IPF.
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divergent expression of Claudin 1 3 4 5 and 7 in developing human lung
Respiratory Research, 2010Co-Authors: Siri Lehtonen, Heta Merikallio, Riitta Kaarteenaho, Terttu Harju, Ylermi SoiniAbstract:Claudins are the main components of tight junctions, structures which are associated with cell polarity and permeability. The aim of this study was to analyze the expression of Claudins 1, 3, 4, 5, and 7 in developing human lung tissues from 12 to 40 weeks of gestation. 47 cases were analyzed by immunohistochemisty for Claudins 1, 3, 4, 5 and 7. 23 cases were also investigated by quantitative RT-PCR for Claudin-1, -3 and -4. Claudin-1 was expressed in epithelium of bronchi and large bronchioles from week 12 onwards but it was not detected in epithelium of developing alveoli. Claudin-3, -4 and -7 were strongly expressed in bronchial epithelium from week 12 to week 40, and they were also expressed in alveoli from week 16 to week 40. Claudin-5 was expressed strongly during all periods in endothelial cells. It was expressed also in epithelium of bronchi from week 12 to week 40, and in alveoli during the canalicular period. RT-PCR analyses revealed detectable amounts of RNAs for Claudins 1, 3 and 4 in all cases studied. Claudin-1, -3, -4, -5, and -7 are expressed in developing human lung from week 12 to week 40 with distinct locations and in divergent quantities. The expression of Claudin-1 was restricted to the bronchial epithelium, whereas Claudin-3, -4 and -7 were positive also in alveolar epithelium as well as in the bronchial epithelium. All Claudins studied are linked to the development of airways, whereas Claudin-3, -4, -5 and -7, but not Claudin-1, are involved in the development of acinus and the differentiation of alveolar epithelial cells.
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Claudins 1 3 4 5 and 7 in esophageal cancer loss of Claudin 3 and 4 expression is associated with metastatic behavior
Apmis, 2007Co-Authors: Heikki Takala, Juha Saarnio, H Wiik, Ylermi SoiniAbstract:The aim of the study was to elucidate the significance of Claudins in surgically treated esophageal carcinoma. The expression of Claudins 1, 3, 4, 5 and 7 was studied by immunohistochemistry. Tumor proliferation was assessed with Ki67 immunostaining and apoptosis by the TUNEL method and immunostaining of fragmented caspase 3. Adenocarcinomas showed significantly more cases with moderate or strong Claudin 3 (p<0.001) and Claudin 5 positivity (p=0.031) compared to squamous cell carcinomas. Loss of Claudin 3 expression was associated with the presence of distant metastases (p=0.039). Claudins 3, 4 and 7 had a significant association with either a high apoptotic index or a high number of caspase 3-positive cells, while Claudin 5 was associated with increased proliferation. In esophageal carcinoma, Claudin expression may vary along with the histology of the tumor. Claudin expression may also be associated with apoptosis or proliferation, suggesting that Claudins may contribute to tumor behavior and growth.
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Claudins 1 3 4 and 5 in gastric carcinoma loss of Claudin expression associates with the diffuse subtype
Virchows Archiv, 2006Co-Authors: Ylermi Soini, S Tommola, Heikki Helin, Paula M MartikainenAbstract:In this study expression of Claudins 1, 3, 4 and 5 were studied in 118 cases of gastric carcinoma and compared with proliferation, apoptosis and E-cadherin expression. Expression of all these Claudins could be seen in gastric carcinoma, most prominently for Claudin 4, and least expression was found for Claudin 5. All Claudins showed significantly more expression in gastric carcinomas of intestinal type. Their expression was significantly associated with each other. Expression of Claudins 4 and 5 was associated with E-cadherin. Strong expression of Claudin 5 was associated with higher cell proliferation and apoptosis. Claudin 3 expression had an association with a better prognosis of the patients, especially in the intestinal type. The results show that expression of Claudins 1, 3, 4 and 5 is lower in diffuse-type gastric carcinomas. Possibly they play a role in determining the diffuse phenotype and loose cohesion of cells in diffuse type of gastric carcinoma in a similar manner as E-cadherin. The loss of their expression does not clearly associate with poorer prognosis of the patients, except for Claudin 3, where strong expression was associated with a better outcome of the patients, a feature especially related to intestinal-type tumours.
Patrice J Morin - One of the best experts on this subject based on the ideXlab platform.
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Claudin proteins in human cancer promising new targets for diagnosis and therapy
Cancer Research, 2005Co-Authors: Patrice J MorinAbstract:The tight junction proteins Claudins are abnormally regulated in several human cancers. In particular, Claudin-3 and Claudin-4 are frequently overexpressed in several neoplasias, including ovarian, breast, pancreatic, and prostate cancers. Although the exact roles of these proteins in tumorigenesis are still being uncovered, it is clear that they represent promising targets for cancer detection, diagnosis, and therapy.
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Claudin 3 and Claudin 4 expression in ovarian epithelial cells enhances invasion and is associated with increased matrix metalloproteinase 2 activity
Cancer Research, 2005Co-Authors: Rachana Agarwal, Theresa Dsouza, Patrice J MorinAbstract:Claudin proteins form a large family of integral membrane proteins crucial for tight junction formation and function. Our previous studies have revealed that Claudin-3 and Claudin-4 proteins are highly overexpressed in ovarian cancer. To clarify the roles of Claudins in ovarian tumorigenesis, we have generated human ovarian surface epithelial (HOSE) cells constitutively expressing wild-type Claudin-3 and Claudin-4. Expression of these Claudins in HOSE cells increased cell invasion and motility as measured by Boyden chamber assays and wound-healing experiments. Conversely, small interfering RNA (siRNA)-mediated knockdown of Claudin-3 and Claudin-4 expression in ovarian cancer cell lines reduced invasion. Claudin expression also increased cell survival in HOSE cells but did not significantly affect cell proliferation. Moreover, the Claudin-expressing ovarian epithelial cells were found to have increased matrix metalloproteinase-2 (MMP-2) activity indicating that Claudin-mediated increased invasion might be mediated through the activation of MMP proteins. However, siRNA inactivation of Claudins in ovarian cancer cell lines did not have a significant effect on the high endogenous MMP-2 activity present in these cells, showing that malignant cells have alternative or additional pathways to fully activate MMP-2. Taken together, our results suggest that Claudin overexpression may promote ovarian tumorigenesis and metastasis through increased invasion and survival of tumor cells.
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phosphorylation of Claudin 3 at threonine 192 by camp dependent protein kinase regulates tight junction barrier function in ovarian cancer cells
Journal of Biological Chemistry, 2005Co-Authors: Theresa Dsouza, Rachana Agarwal, Patrice J MorinAbstract:Abstract Claudins are integral membrane proteins essential in the formation and function of tight junctions (TJs). Disruption of TJs, which have essential roles in cell permeability and polarity, is thought to contribute to epithelial tumorigenesis. Claudin-3 and -4 are frequently overexpressed in ovarian cancer, but the molecular pathways involved in the regulation of these proteins are unclear. Interestingly, several studies have demonstrated a role for phosphorylation in the regulation of TJ complexes, although evidence for Claudin phosphorylation is scarce. Here, we showed that Claudin-3 and -4 can be phosphorylated in ovarian cancer cells. In vitro phosphorylation assays using glutathione S-transferase fusion constructs demonstrated that the C terminus of Claudin-3 is an excellent substrate for cAMP-dependent protein kinase (PKA). Using site-directed mutagenesis, we identified a PKA phosphorylation site at amino acid 192 in the C terminus of Claudin-3. Overexpression of the protein containing a T192D mutation, mimicking the phosphorylated state, resulted in a decrease in TJ strength in ovarian cancer cell line OVCA433. Our results suggest that Claudin-3 phosphorylation by PKA, a kinase frequently activated in ovarian cancer, may provide a mechanism for the disruption of TJs in this cancer. In addition, our findings may have general implications for the regulation of TJs in normal epithelial cells.
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tight junction proteins Claudin 3 and Claudin 4 are frequently overexpressed in ovarian cancer but not in ovarian cystadenomas
Clinical Cancer Research, 2003Co-Authors: Leticia Rangel, Rachana Agarwal, Theresa Dsouza, Ellen S Pizer, Wayne D Lancaster, Lucie Gregoire, Donald R Schwartz, Patrice J MorinAbstract:Purpose: Claudin proteins represent a large family of integral membrane proteins crucial for tight junction (TJ) formation and function. Claudins have been shown to be up-regulated in various cancers and have been suggested as possible biomarkers and targets for cancer therapy. Because Claudin-3 and Claudin-4 have been proposed to be expressed in epithelial ovarian cancer, we have performed a detailed analysis of CLDN3 and CLDN4 expression in a panel of ovarian tumors of various subtypes and cell lines. We also investigated whether high expression of Claudin-3 and Claudin-4 was associated with TJ function in ovarian cancer cells. Experimental Design: RNA was obtained from a panel of 39 microdissected epithelial ovarian tumors of various histological subtypes for real-time reverse transcription-PCR analysis. In addition, a total of 70 cases of ovarian carcinomas, ovarian cysts, and normal ovarian epithelium from a tissue array were analyzed by immunohistochemistry. Finally, a panel of cell lines was used for Western analysis of Claudin expression and TJ permeability studies. Results: Although expressed at low levels in some normal human tissues, including the ovary, CLDN3 and CLDN4 are highly up-regulated in epithelial ovarian cancers of all subtypes. Immunohistochemical analyses using our ovarian tissue array confirmed the high level of expression of Claudin-3 and Claudin-4 in the majority of ovarian carcinomas, including many tumors exhibiting cytoplasmic staining. Ovarian cystadenoma did not frequently overexpress these proteins, suggesting that the expression of these proteins is associated with malignancy. In ovarian cancer cell lines, Claudin-3 and Claudin-4 expression was not associated with functional TJs as measured by transepithelial electrical resistance. Conclusions: These results show that CLDN3 and CLDN4 are frequently up-regulated in ovarian tumors and cell lines and may represent novel markers for this disease. Overexpression of these genes in ovarian cancer also suggests interesting scenarios for the involvement of TJ in tumorigenesis. A better knowledge of the mechanisms underlying ovarian tumorigenesis will likely result in the development of novel approaches for the diagnosis and therapy of this deadly disease.
Theodore G Gabig - One of the best experts on this subject based on the ideXlab platform.
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a Claudin 3 and Claudin 4 targeted clostridium perfringens protoxin is selectively cytotoxic to psa producing prostate cancer cells
Cancer Letters, 2014Co-Authors: Victor Romanov, Terry Whyard, Wayne C Waltzer, Theodore G GabigAbstract:Abstract Prostate cancer is the second leading cause of non-cutaneous cancer-related death in males, and effective strategies for treatment of metastatic disease are currently limited. The tight junction proteins, Claudin 3 and Claudin 4, serve as cell-surface receptors for the pore-forming Clostridium perfringens enterotoxin [CPE]. Most prostate cancer cells overexpress Claudin 3 and Claudin 4, and Claudins are aberrantly distributed over the plasma membrane, making these cells particularly sensitive to cytolysis by CPE. Prostate cancer cells secrete PSA locally that is proteolytically active; however, circulating PSA is inactivated via binding to protease inhibitors. To overcome systemic toxicity of CPE, a modified protoxin was constructed with a tethered ligand attached to the C-terminus connected by a flexible linker containing a PSA-specific protease cleavage site. This engineered protoxin selectively and efficiently lyses PSA-producing prostate cancer cells whereas CLDN3 and CLDN4 positive cells that do not express PSA are resistant to cytolysis.
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clostridium perfringens enterotoxin elicits rapid and specific cytolysis of breast carcinoma cells mediated through tight junction proteins Claudin 3 and 4
American Journal of Pathology, 2004Co-Authors: Scott L Kominsky, Theodore G Gabig, Mustafa Vali, Dorian Korz, Sigmund A Weitzman, Pedram Argani, Saraswati SukumarAbstract:Clostridium perfringens enterotoxin (CPE) induces cytolysis very rapidly through binding to its receptors, the tight junction proteins CLDN 3 and 4. In this study, we investigated CLDN 3 and 4 expression in breast cancer and tested the potential of CPE-mediated therapy. CLDN 3 and 4 proteins were detected in all primary breast carcinomas tested (n = 21) and, compared to normal mammary epithelium, were overexpressed in approximately 62% and 26%, respectively. Treatment of breast cancer cell lines in culture with CPE resulted in rapid and dose-dependent cytolysis exclusively in cells that expressed CLDN 3 and 4. Intratumoral CPE treatment of xenografts of T47D breast cancer cells in immunodeficient mice resulted in a significant reduction in tumor volume (P = 0.007), with accompanying necrosis. Necrotic reactions were also seen in three freshly resected primary breast carcinoma samples treated with CPE for 12 hours, while isolated primary breast carcinoma cells underwent rapid and complete cytolysis within 1 hour. Thus, expression of CLDN 3 and 4 sensitizes primary breast carcinomas to CPE-mediated cytolysis and emphasizes the potential of CPE in breast cancer therapy.
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clostridium perfringens enterotoxin elicits rapid and specific cytolysis of breast carcinoma cells mediated through tight junction proteins Claudin 3 and 4
American Journal of Pathology, 2004Co-Authors: Scott L Kominsky, Theodore G Gabig, Mustafa Vali, Dorian Korz, Sigmund A Weitzman, Pedram Argani, Saraswati SukumarAbstract:Clostridium perfringens enterotoxin (CPE) induces cytolysis very rapidly through binding to its receptors, the tight junction proteins CLDN 3 and 4. In this study, we investigated CLDN 3 and 4 expression in breast cancer and tested the potential of CPE-mediated therapy. CLDN 3 and 4 proteins were detected in all primary breast carcinomas tested (n = 21) and, compared to normal mammary epithelium, were overexpressed in approximately 62% and 26%, respectively. Treatment of breast cancer cell lines in culture with CPE resulted in rapid and dose-dependent cytolysis exclusively in cells that expressed CLDN 3 and 4. Intratumoral CPE treatment of xenografts of T47D breast cancer cells in immunodeficient mice resulted in a significant reduction in tumor volume (P = 0.007), with accompanying necrosis. Necrotic reactions were also seen in three freshly resected primary breast carcinoma samples treated with CPE for 12 hours, while isolated primary breast carcinoma cells underwent rapid and complete cytolysis within 1 hour. Thus, expression of CLDN 3 and 4 sensitizes primary breast carcinomas to CPE-mediated cytolysis and emphasizes the potential of CPE in breast cancer therapy.
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expression of clostridium perfringens enterotoxin receptors Claudin 3 and Claudin 4 in prostate cancer epithelium
Cancer Research, 2001Co-Authors: Haiyan Long, Colin D Crean, Weihua Lee, William O Cummings, Theodore G GabigAbstract:The mRNA for Rvp.1 (rat ventral prostate) increases in abundance before gland involution after androgen deprivation. Rvp.1 is homologous to CPE-R, the high-affinity intestinal epithelial receptor for Clostridium perfringens enterotoxin (CPE), and is sufficient to mediate CPE binding and trigger subsequent toxin-mediated cytolysis. Rvp.1 (Claudin-3) and CPE-R (Claudin-4) are members of a larger family of transmembrane tissue-specific Claudin proteins that are essential components of intercellular tight junction structures regulating paracellular ion flux. However, Claudin-3 and Claudin-4 are the only family members capable of mediating CPE binding and cytolysis. The present study was designed to study the expression of Claudin-3 and Claudin-4 in human prostate tissue as potential targets for CPE toxin-mediated therapy for prostate cancer. On human multiple-tissue Northern blot analysis, mRNAs for both Claudin-3 and Claudin-4 were expressed at high levels in prostate tissue. In normal prostate tissue, expression of Claudin-3 was localized exclusively within acinar epithelial cells by in situ mRNA hybridization. Compared with expression within prostate epithelial cells in surrounding normal glandular tissue, expression of Claudin-3 mRNA remained high in the epithelium of prostate adenocarcinoma (10 of 10) and prostatic intraepithelial neoplasia (five of five). Prostate adenocarcinoma cells metastatic to bone were obtained from a patient with disease progression during antiandrogen therapy. These metastatic cells were prostate-specific antigen-positive by immunohistochemical staining and also expressed functional CPE receptors as measured by sensitivity to CPE-induced cell lysis. The persistent high level of Claudin-3 expression in prostate adenocarcinoma and functional cytotoxicity of CPE in metastatic androgen-independent prostate adenocarcinoma suggests a new potential therapeutic strategy for prostate cancer.
Alessandro D Santin - One of the best experts on this subject based on the ideXlab platform.
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clostridium perfringens enterotoxin cpe and cpe binding domain c cpe for the detection and treatment of gynecologic cancers
Toxins, 2015Co-Authors: Jonathan Black, Salvatore Lopez, Emiliano Cocco, Carlton L Schwab, Diana P English, Alessandro D SantinAbstract:Clostridium perfringens enterotoxin (CPE) is a three-domain polypeptide, which binds to Claudin-3 and Claudin-4 with high affinity. Because these receptors are highly differentially expressed in many human tumors, Claudin-3 and Claudin-4 may provide an efficient molecular tool to specifically identify and target biologically aggressive human cancer cells for CPE-specific binding and cytolysis. In this review we will discuss these surface proteins as targets for the detection and treatment of chemotherapy-resistant gynecologic malignancies overexpressing Claudin-3 and -4 using CPE-based theranostic agents. We will also discuss the use of fluorescent c-CPE peptide in the operative setting for real time detection of micro-metastatic tumors during surgery and review the potential role of CPE in other medical applications.
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overexpression of Claudin 3 and Claudin 4 receptors in uterine serous papillary carcinoma novel targets for a type specific therapy using clostridium perfringens enterotoxin cpe
Cancer, 2007Co-Authors: Alessandro D Santin, Stefania Bellone, Moira Marizzoni, Michela Palmieri, Eric R Siegel, Jesse K Mckenney, Leah Hennings, Fabrizio Comper, Elisabetta Bandiera, Sergio PecorelliAbstract:BACKGROUND. Uterine serous papillary carcinoma (USPC) represents a highly aggressive variant of endometrial cancer. Using gene expression profiling, we recently identified high expression of the Claudin-3 and Claudin-4 receptors in a limited set of USPC. These tight junction proteins represent the low- and high-affinity receptors, respectively, for the cytotoxic Clostridium perfringens enterotoxin (CPE) and are sufficient to mediate CPE binding and trigger subsequent toxin-mediated cytolysis. The potential for targeting this pathway in the treatment of USPC was explored. METHODS. Claudin-3 and Claudin-4 receptor expression was analyzed at the mRNA and protein levels in flash-frozen and formalin-fixed, paraffin-embedded tissue from 20 consecutive USPC patients. The potential of recombinant CPE as a novel therapy against primary, metastatic, and chemotherapy-resistant USPC cell lines was also investigated in vitro. Finally, the in vivo therapeutic effect of sublethal doses of CPE was studied in SCID mouse xenografts harboring subcutaneous and intraperitoneal USPC that expressed Claudin-3 and Claudin-4. RESULTS. In all, 100% (20 out of 20) of the primary flash-frozen USPC tested overexpressed 1 or both CPE receptors by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR). Membranous immunoreactivity for Claudin-4 protein expression was documented in the majority of USPC specimens tested by immunohistochemistry, whereas only a low level of membranous staining was found in normal endometrial control tissue samples. When primary and metastatic short-term USPC cell lines were incubated with different concentrations of CPE in vitro, a dose-dependent cytotoxic effect was demonstrated. In vivo, intratumoral injections of well-tolerated doses of CPE in large subcutaneous USPC xenografts led to large areas of tumor cell necrosis and tumor disappearance in all the treated animals, whereas sublethal intraperitoneal injections of CPE had a significant inhibitory effect on tumor progression, with extended survival of animals harboring chemotherapy-resistant intra-abdominal USPC carcinomatosis. CONCLUSIONS. Claudin-3 and Claudin-4 receptors may offer promising targets for the use of CPE as a novel type-specific therapy against this highly aggressive and chemotherapy-resistant variant of endometrial cancer. Cancer 2007. © 2007 American Cancer Society.
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treatment of chemotherapy resistant human ovarian cancer xenografts in c b 17 scid mice by intraperitoneal administration of clostridium perfringens enterotoxin
Cancer Research, 2005Co-Authors: Alessandro D Santin, Stefania Bellone, Michela Palmieri, Eric R Siegel, Stefania Cane, Maria Thomas, Juan J Roman, Alexander F Burnett, Martin J Cannon, Sergio PecorelliAbstract:Ovarian cancer remains the most lethal gynecologic malignancy in the United States. Although many patients with advanced-stage disease initially respond to standard combinations of surgical and cytotoxic therapy, nearly 90% develop recurrence and inevitably die from the development of chemotherapy-resistant disease. The discovery of novel and effective therapy against chemotherapy-resistant/recurrent ovarian cancer remains a high priority. Using expression profiling, we and others have recently found Claudin-3 and Claudin-4 genes to be highly expressed in ovarian cancer. Because these tight junction proteins have been described as the low- and high-affinity receptors, respectively, for the cytotoxic Clostridium perfringens enterotoxin (CPE), in this study we investigated the level of expression of Claudin-3 and/or Claudin-4 in chemotherapy-naive and chemotherapy-resistant primary human ovarian cancers as well as their sensitivity to CPE treatment in vitro. We report that 100% (17 of 17) of the primary ovarian tumors tested overexpress one or both CPE receptors by quantitative reverse transcription-PCR. All ovarian tumors showed a dose-dependent cytotoxic effect to CPE in vitro. Importantly, chemotherapy-resistant/recurrent ovarian tumors were found to express Claudin-3 and Claudin-4 genes at significantly higher levels when compared with chemotherapy-naive ovarian cancers. All primary ovarian tumors tested, regardless of their resistance to chemotherapeutic agents, died within 24 hours to the exposure to 3.3 microg/mL CPE in vitro. In addition, we have studied the in vivo efficacy of i.p. CPE therapy in SCID mouse xenografts in a highly relevant clinical model of chemotherapy-resistant freshly explanted human ovarian cancer (i.e., OVA-1). Multiple i.p. administration of sublethal doses of CPE every 3 days significantly inhibited tumor growth in 100% of mice harboring 1 week established OVA-1. Repeated i.p. doses of CPE also had a significant inhibitory effect on tumor progression with extended survival of animals harboring large ovarian tumor burdens (i.e., 4-week established OVA-1). Our findings suggest that CPE may have potential as a novel treatment for chemotherapy-resistant/recurrent ovarian cancer.
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gene expression profiles in primary ovarian serous papillary tumors and normal ovarian epithelium identification of candidate molecular markers for ovarian cancer diagnosis and therapy
International Journal of Cancer, 2004Co-Authors: Alessandro D Santin, Stefania Bellone, Michela Palmieri, Stefania Cane, Fenghuang Zhan, Eliana Bignotti, Simone Anfossi, Murat Gokden, Donna Dunn, Juan J RomanAbstract:With the goal of identifying genes with a differential pattern of expression between ovarian serous papillary carcinomas (OSPCs) and normal ovarian (NOVA) epithelium and using this knowledge for the development of novel diagnostic and therapeutic markers for ovarian cancer, we used oligonucleotide microarrays with probe sets complementary to 12,533 genes to analyze the gene expression profiles of 10 primary OSPC cell lines, 2 established OSPC cell lines (UCI-101, UCI-107) and 5 primary NOVA epithelial cultures. Unsupervised analysis of gene expression data identified 129 and 170 genes that exhibited >5-fold upregulation and downregulation, respectively, in primary OSPC compared to NOVA. Genes overexpressed in established OSPC cell lines had little correlation with those overexpressed in primary OSPC, highlighting the divergence of gene expression that occurs as a result of long-term in vitro growth. Hierarchical clustering of the expression data readily distinguished normal tissue from primary OSPC. Laminin, Claudin 3, Claudin 4, tumor-associated calcium signal transducers 1 and 2 (TROP-1/Ep-CAM, TROP-2), ladinin 1, S100A2, SERPIN2 (PAI-2), CD24, lipocalin 2, osteopontin, kallikrein 6 (protease M), kallikrein 10, matriptase (TADG-15) and stratifin were among the most highly overexpressed genes in OSPC compared to NOVA. Downregulated genes in OSPC included transforming growth factor-beta receptor III, platelet-derived growth factor receptor alpha, SEMACAP3, ras homolog gene family member I (ARHI), thrombospondin 2 and disabled-2/differentially expressed in ovarian carcinoma 2 (Dab2/DOC2). Differential expression of some of these genes, including Claudin 3, Claudin 4, TROP-1 and CD24, was validated by quantitative RT-PCR and flow cytometry on primary OSPC and NOVA. Immunohistochemical staining of formalin-fixed, paraffin-embedded tumor specimens from which primary OSPC cultures were derived further confirmed differential expression of CD24 and TROP-1/Ep-CAM markers on OSPC vs. NOVA. These results, obtained with highly purified primary cultures of ovarian cancer, highlight important molecular features of OSPC and may provide a foundation for the development of new type-specific therapies against this disease.
Saraswati Sukumar - One of the best experts on this subject based on the ideXlab platform.
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clostridium perfringens enterotoxin elicits rapid and specific cytolysis of breast carcinoma cells mediated through tight junction proteins Claudin 3 and 4
American Journal of Pathology, 2004Co-Authors: Scott L Kominsky, Theodore G Gabig, Mustafa Vali, Dorian Korz, Sigmund A Weitzman, Pedram Argani, Saraswati SukumarAbstract:Clostridium perfringens enterotoxin (CPE) induces cytolysis very rapidly through binding to its receptors, the tight junction proteins CLDN 3 and 4. In this study, we investigated CLDN 3 and 4 expression in breast cancer and tested the potential of CPE-mediated therapy. CLDN 3 and 4 proteins were detected in all primary breast carcinomas tested (n = 21) and, compared to normal mammary epithelium, were overexpressed in approximately 62% and 26%, respectively. Treatment of breast cancer cell lines in culture with CPE resulted in rapid and dose-dependent cytolysis exclusively in cells that expressed CLDN 3 and 4. Intratumoral CPE treatment of xenografts of T47D breast cancer cells in immunodeficient mice resulted in a significant reduction in tumor volume (P = 0.007), with accompanying necrosis. Necrotic reactions were also seen in three freshly resected primary breast carcinoma samples treated with CPE for 12 hours, while isolated primary breast carcinoma cells underwent rapid and complete cytolysis within 1 hour. Thus, expression of CLDN 3 and 4 sensitizes primary breast carcinomas to CPE-mediated cytolysis and emphasizes the potential of CPE in breast cancer therapy.
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clostridium perfringens enterotoxin elicits rapid and specific cytolysis of breast carcinoma cells mediated through tight junction proteins Claudin 3 and 4
American Journal of Pathology, 2004Co-Authors: Scott L Kominsky, Theodore G Gabig, Mustafa Vali, Dorian Korz, Sigmund A Weitzman, Pedram Argani, Saraswati SukumarAbstract:Clostridium perfringens enterotoxin (CPE) induces cytolysis very rapidly through binding to its receptors, the tight junction proteins CLDN 3 and 4. In this study, we investigated CLDN 3 and 4 expression in breast cancer and tested the potential of CPE-mediated therapy. CLDN 3 and 4 proteins were detected in all primary breast carcinomas tested (n = 21) and, compared to normal mammary epithelium, were overexpressed in approximately 62% and 26%, respectively. Treatment of breast cancer cell lines in culture with CPE resulted in rapid and dose-dependent cytolysis exclusively in cells that expressed CLDN 3 and 4. Intratumoral CPE treatment of xenografts of T47D breast cancer cells in immunodeficient mice resulted in a significant reduction in tumor volume (P = 0.007), with accompanying necrosis. Necrotic reactions were also seen in three freshly resected primary breast carcinoma samples treated with CPE for 12 hours, while isolated primary breast carcinoma cells underwent rapid and complete cytolysis within 1 hour. Thus, expression of CLDN 3 and 4 sensitizes primary breast carcinomas to CPE-mediated cytolysis and emphasizes the potential of CPE in breast cancer therapy.