No abstract available.
Blood Platelets* ; Hypertension, Pulmonary* ; Lung Diseases, Obstructive* ; Platelet Activation*

PURPOSE: Both genetic and epigenetic alterations can lead to abnormal expression of metastasis-regulating genes in tumor cells. Recent studies suggest that aberrant epigenetic alterations, followed by differential gene expression, leads to an aggressive cancer cell phenotype. We examined epigenetically regulated genes that are involved in ovarian cancer metastasis. MATERIALS AND METHODS: We developed SK-OV-3 human ovarian carcinoma cell xenografts in mice. We compared the mRNA expression and DNA methylation profiles of metastatic tissues to those of the original SK-OV-3 cell line. RESULTS: Metastatic implants showed increased mRNA expression of the carbonic anhydrase 9 (CA9) gene and hypomethylation at CpG sites in the CA9 promoter. Treatment of wild-type SK-OV-3 cells with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine reduced methylation of the CA9 promoter and increased CA9 mRNA expression. Eight CpGs, which were located at positions -197, -74, -19, -6, +4, +13, +40, and +86, relative to the transcription start site, were hypomethylated in metastatic tumor implants, compared to that of wild-type SK-OV-3. Overexpression of CA9 induced an aggressive phenotype, including increased invasiveness and migration, in SK-OV-3 cells. CONCLUSION: Alterations in the DNA methylation profile of the CA9 promoter were correlated with a more aggressive phenotype in ovarian cancer cells.
Animals ; Azacitidine/*analogs & derivatives/pharmacology ; Carbonic Anhydrases/metabolism ; *DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic/*drug effects ; Humans ; Mice ; Neoplasm Invasiveness/genetics ; Neoplasm Metastasis/genetics/*pathology ; Neoplasms, Experimental ; Neoplasms, Glandular and Epithelial/genetics/*metabolism/pathology ; Ovarian Neoplasms/genetics/*metabolism/pathology ; Phenotype ; Promoter Regions, Genetic ; RNA, Messenger/metabolism

PURPOSE: Glioblastoma multiforme (GBM) is the most common adult primary intracranial tumor. The remarkable features of GBM include central necrosis. MicroRNAs (miRNAs) have been considered as diagnostic/prognostic biomarkers for many cancers, including glioblastoma. However, the effect of necrosis on the miRNA expression profile and predicted miRNA-mRNA regulatory information remain unclear. The purpose of this study is to examine the effect of necrotic cells on the modulation of miRNA and mRNA expression profiles and miRNA-mRNA network in CRT-MG cells. MATERIALS AND METHODS: We used human astroglioma cells, CRT-MG, treated with necrotic CRT-MG cells to examine the effect of necrosis on the modulation of miRNA and mRNA by next-generation sequencing. For preparation of necrotic cells, CRT-MG cells were frozen and thawed through cycle of liquid nitrogen–water bath. The putative miRNA-mRNA regulatory relationship was inferred through target information, using miRDB. RESULTS: The necrotic cells induced dysregulation of 106 miRNAs and 887 mRNAs. Among them, 11 miRNAs that had a negative correlation value of p < 0.05 by the hypergeometric test were screened, and their target mRNAs were analyzed by Gene Ontology enrichment analysis. Using the Kyoto Encyclopedia of Genes and Genomes database, we also found several necrotic cell treatment-activated pathways that were modulated by relevant gene targets of differentially expressed miRNAs. CONCLUSION: Our result demonstrated that dysregulation of miRNA and mRNA expression profiles occurs when GBM cells are exposed to necrotic cells, suggesting that several miRNAs may have the potential to be used as biomarkers for predicting GBM progression and pathogenesis.
Adult ; Astrocytoma* ; Baths ; Biomarkers ; Gene Ontology ; Genome ; Glioblastoma ; Humans* ; MicroRNAs ; Necrosis* ; RNA, Messenger

BACKGROUND: Post-pneumonectomy empyema(PPE) is an uncommon but a serious complication. The management remains as challenge for general thoracic surgeons. MATERIAL AND METHOD: During the period of January 1990 to December 1996, we evaluated the results of 20 patients with post-pneumonectomy empyema. RESULT: Sex ratio were 15 male and 5 female patients with mean age of 41.5+/-21.5 yrs. The occurrence ratio of left to right side was 8:12. The most common disease for prior pneumonectomy was pulmonary tuberculosis. The duration between pneumonectomy and PPE was variable in 1 month to 6yrs. Fever was the most frequent symptom and S. aureus was the most frequent pathogen. In 13 cases, there were combined with BPF. Four patients underwent trans-sternal closure, and Clagett procedure was performed. There was one recurrence that later underwent muscle plombage and omentopexy later. Nine patients underwent omentopexy, muscle plombage and thoracoplasty. There were 7 cases that were not combined with BPF. All 7 patients underwent thoracoplasty, and two of them were combined with muscle plombage. Mean follow-up duration is 40+/-32.3 months. There were no late deaths nor recurrences of PPE. CONCLUSION: We conclude that early diagnosis and proper drainage in PPE patients are important in its initial stage of management, and also management is completely achieved in thoracoplasty with muscle plombage or omentopexy.
Drainage ; Early Diagnosis ; Empyema* ; Female ; Fever ; Follow-Up Studies ; Humans ; Male ; Pneumonectomy ; Recurrence ; Sex Ratio ; Thoracoplasty ; Tuberculosis, Pulmonary

PURPOSE: Ovarian cancer (OC) is the most fatal of gynecological malignancies with a high rate of recurrence. We aimed to evaluate the expression of solute carrier family 6, member 12 (SLC6A12) and methylation of its promoter CpG sites in a xenograft mouse model of metastatic OC, and to investigate the regulatory mechanisms that promote aggressive properties during OC progression. MATERIALS AND METHODS: Expression of SLC6A12 mRNA was determined by reverse-transcription quantitative polymerase chain reaction (RT-qPCR), and DNA methylation status of its promoter CpGs was detected by quantitative methylation-specific PCR. The metastatic potential of SLC6A12 was evaluated by in vitro migration/invasion transwell assays. Gene expression and DNA methylation of SLC6A12 and clinical outcomes were further investigated from publicly available databases from curatedOvarianData and The Cancer Genome Atlas. RESULTS: SLC6A12 expression was 8.1–14.0-fold upregulated and its DNA methylation of promoter CpG sites was 41–62% decreased in tumor metastases. After treatment with DNA methyltransferase inhibitor and/or histone deacetylase inhibitor, the expression of SLC6A12 was profoundly enhanced (~8.0-fold), strongly supporting DNA methylation-dependent epigenetic regulation of SLC6A12. Overexpression of SLC6A12 led to increased migration and invasion of ovarian carcinoma cells in vitro, approximately 2.0-fold and 3.3-fold, respectively. The meta-analysis showed that high expression of SLC6A12 was significantly associated with poor overall survival [hazard ratio (HR)=1.07, p value=0.016] and that low DNA methylation levels of SLC6A12 at specific promoter CpG site negatively affected patient survival. CONCLUSION: Our findings provide novel evidence for the biological and clinical significance of SLC6A12 as a metastasis-promoting gene.
Animals ; Carrier Proteins/genetics/*metabolism ; Cell Line, Tumor ; Cell Migration Assays ; *CpG Islands ; *DNA Methylation ; Disease Progression ; Epigenesis, Genetic ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Ovarian Neoplasms/genetics/*metabolism/mortality/pathology ; Polymerase Chain Reaction ; Prognosis ; *Promoter Regions, Genetic ; RNA, Messenger/*metabolism ; Up-Regulation

Intravenous leiomyomatosis (IVL) is a rare neoplasm that is characterized by a histologically benigh-looking smooth muscle cell tumor growing within the uterine and the extrauterine venous system. We report here on patient had a uterine and extrauterine leiomyoma that extended into the right atrium through the inferior vena cava. A 19-year old female patient was suffering from dyspnea, swelling of the lower extremity, abdominal pain and abdominal distension. She had total abdominal hysterectomy performed at a private clinic due to uterine leiomyoma 4 months previously. 4 months after the first operation, we again completely excised the recurred intraperitoneal tumor mass. At 6 months after the re-operation, the tumor mass recurred intraperitoneally. A preoperative abdominal CT scan and an echocardiogram revealed multiple tumor masses that were located intraperitoneally and they extended to the right atrium. We performed intraperitoneal tumor excision and removal of the intravenous tumor mass via the right iliac vein by the one-stage approach. At present, the patient has shown a clinically favorable outcome except for local recurrence of tumor mass in the pelvic cavity. The cure of this complex disease emphasizes the need for a planned systemic approach by a multidisciplinary surgical team. We present here an unusual case of intravenous leiomyomatosis that originated from the uterus and it extended to the inferior vena cava and right atrium.
Abdominal Pain ; Dyspnea ; Female ; Heart Atria ; Humans ; Hysterectomy ; Iliac Vein ; Leiomyoma ; Leiomyomatosis* ; Lower Extremity ; Myocytes, Smooth Muscle ; Recurrence ; Tomography, X-Ray Computed ; Uterus ; Vena Cava, Inferior ; Young Adult

Metastasis is a major cause of therapeutic failure in ovarian cancer. To elucidate molecular mechanisms of ovarian cancer metastasis, we previously established a metastatic xenograft mouse model using human ovarian carcinoma SK-OV-3 cells. Using gene expression profiling, we found that γ-aminobutyric acid (GABA)A receptor π subunit (GABRP) expression was upregulated (>4-fold) in metastatic tissues from our xenograft mice compared with SK-OV-3 cells. Importantly, GABRP knockdown diminished the migration and invasion of SK-OV-3 cells, and reduced extracellular signal-regulated kinase (ERK) activation while overexpression of GABRP exhibited significantly increased cell migration, invasion and ERK activation. Moreover, treatment with the mitogen-activated protein kinase (MAPK)/ERK kinase (MEK) inhibitor U0126 similarly suppressed the migration and invasion of SK-OV-3 cells, implying that GABRP promotes these cellular behaviors by activating the MAPK/ERK pathway. Using genome-wide DNA methylation profiling, we identified hypomethylated CpG sites in the GABRP promoter in metastatic tissues from the xenograft mice compared with SK-OV-3 cells. Treatment with a DNA methyltransferase inhibitor demonstrated that methylation at −963 bp from the GABRP transcription start site (−963 CpG site) was critical for the epigenetic regulation of GABRP. Finally, we analyzed human ovarian cancer patient samples and showed DNA hypomethylation at the GABRP −963 CpG site in advanced stage, but not early-stage, primary tumors compared with their paired normal tissues. These findings suggest that GABRP enhances the aggressive phenotype of ovarian cancer cells, and that the DNA methylation status of the GABRP −963 CpG site may be useful for predicting the metastatic potential in ovarian cancer patients.
Animals ; Cell Movement ; DNA ; DNA Methylation ; Epigenomics* ; Gene Expression Profiling ; Heterografts ; Humans ; Methylation ; Mice ; Neoplasm Metastasis ; Ovarian Neoplasms* ; Phenotype* ; Phosphotransferases ; Protein Kinases ; Transcription Initiation Site

PURPOSE: Idoxifene is currently entering phase II clinical trials for the treatment of advanced breast cancer. The radiolabeled idoxifene using 123I provides an opportunity for clinical pharmacology with single photon emission computed tomography (SPECT). The purpose of this study was to prepare radiolabeled idoxifene using 123I and to determine its cell uptake of breast cancer cell line. MATERIALS AND METHODS: With a view to evaluating new anticancer drugs, we are investigating the novel antiestrogen pyrrolidino- 4-iodotamoxifen (idoxifene). [123I]Idoxifene has been prepared in no-carrier-added form using a tributyl stannylated precursor which has been synthesized by means of (2-chloroethoxy)benzene with (+/-)-2- phenylbutanoic acid on the basis of previously reported standard methods. The biodistribution and dynamic behavior of the compound were investigated using the comparative breast cancer cell line, MCF-7 (estrogen receptor-positive) and MDA-MB-468 (non-estrogen receptor). RESULTS AND CONCLUSION: Acylation of (2-chloroethoxy)benzene with (+/-)-2-phenylbutanoic acid gave the versatile ketone (81%) which reacted with 1,4-diiodobenzene to give triphenylethylene as a mixture of E and Z geometric isomers, which were separated by the recrystallization in ethanol. The E-isomer was treated with pyrrolidine to give idoxifene (67%). In order to incorporate radioactive iodine into the 4-position, the 4-stannylated precursor was prepared (30%). The yield of radioiodination was 90-92% with a high radiochemical purity greater than 98%. The ratio of tumor uptake of the breast cancer cell line between MCF-7 and MDA-MB-468 was about 1.7.
Acylation ; Breast Neoplasms* ; Breast* ; Cell Line ; Estrogen Receptor Modulators ; Ethanol ; Iodine ; Pharmacology, Clinical ; Tomography, Emission-Computed, Single-Photon

OBJECTIVE: The pathologic diagnosis of the astrocytoma has been primarily based on the histologic grading, however, there are some discrepancies among the pathologists on the tumor grading. Met protein, known as the hepatocyte growth factor receptor, is a transmembrane 190 kDa heterodimer with tyrosine kinase activity, encoded by c-met gene. Although c-Met protein is known to be expressed in a variety of tissues and plays important roles in signal transduction, the study on its expression related to clinicopathological prognostic parameters in brain tumor is rare. METHODS: We have evaluated c-Met protein expression in association with p53 and Ki-67 expression in 35 astrocytic tumors (15 diffuse astrocytomas: LGA, 11 anaplastic astrocytomas: AA, 9 Glioblastoma multiforme: GBM) using immunohistochemical method. RESULTS: c-Met immunoreactivity was observed in 2 LGA(13.3%), 5AA(45.5%), 4GBM cases (44.4%), respectively. p53 immunoreactivity was observed in 2 LGA(13.3%), 4AA(36.4%), 4GBM cases (44.4%), respectively. Ki-67 labelling index was 1.7+/-1.0% (LGA), 13.3+/-.2% (AA) and 18.0+/-.1% (GBM), respectively. Each c-Met expression and the Ki-67 labelling index were statistically correlated between low grade and anaplastic astrocytomas. The c-Met and p53 expression rate were not associated with increased Ki-67 labelling index. But, c-Met, p53, Ki-67 expression tended to increase with higher grade of malignancy. CONCLUSION: We conclude that c-Met expression may contribute to the invasiveness and tumor progression of the astrocytoma and c-Met expression is useful in discrimination between low grade astrocytoma and anaplastic astrocytoma.
Astrocytoma ; Brain Neoplasms ; Diagnosis ; Discrimination (Psychology) ; Glioblastoma ; Neoplasm Grading ; Protein-Tyrosine Kinases ; Proto-Oncogene Proteins c-met ; Signal Transduction

PURPOSE: Recent discoveries suggest that aberrant DNA methylation provides cancer cells with advanced metastatic properties. However, the precise regulatory mechanisms controlling metastasis genes and their role in metastatic transformation are largely unknown. To address epigenetically-regulated gene products involved in ovarian cancer metastasis, we examined the mechanisms regulating mucin 13 (MUC13) expression and its influence on aggressive behaviors of ovarian malignancies. MATERIALS AND METHODS: We injected SK-OV-3 ovarian cancer cells peritoneally into nude mice to mimic human ovarian tumor metastasis. Overexpression of MUC13 mRNA was detected in metastatic implants from the xenografts by expression microarray analysis and quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The DNA methylation status within the MUC13 promoter region was determined using bisulfite sequencing PCR and quantitative methylation-specific PCR. We evaluated the effects of exogenous MUC13 on cell invasion and migration using in vitro transwell assays. RESULTS: MUC13 mRNA expression was up-regulated, and methylation of specific CpG sites within the promoter was reduced in the metastatic implants relative to those in wild-type SK-OV-3 cells. Addition of a DNA methyltransferase inhibitor to SK-OV-3 cells induced MUC13 expression, thereby implying epigenetic regulation of MUC13 by promoter methylation. MUC13 overexpression increased migration and invasiveness, compared to control cells, suggesting aberrant up-regulation of MUC13 is strongly associated with progression of aggressive behaviors in ovarian cancer. CONCLUSION: We provide novel evidence for epigenetic regulation of MUC13 in ovarian cancer. We suggest that the DNA methylation status within the MUC13 promoter region may be a potential biomarker of aggressive behavior in ovarian cancer.
Animals ; Cell Line, Tumor ; *DNA Methylation ; Epigenesis, Genetic ; Female ; *Gene Expression Regulation, Neoplastic ; Heterografts/metabolism ; Humans ; Mice ; Mice, Nude ; Mucins/*genetics/metabolism/physiology ; Neoplasm Invasiveness/genetics ; Ovarian Neoplasms/genetics/*metabolism/pathology ; RNA, Messenger/metabolism