OBJECTIVETo study the role of arginase-1 (Arg-1) expression in differential diagnosis of hepatocellular carcinoma (HCC), Arg-1 staining pattern in clear cell neoplasm (HCC and non-HCC) and Arg-1 expression in non-hepatocellular tumors.

METHODSSeventy-eight cases of HCC (including 8 cases of clear cell type and 70 cases of non- clear cell type) and 246 cases of non-hepatocellular neoplasms (including 29 cases of metastatic tumors such as breast cancer, nasopharyngeal carcinoma and neuroendocrine carcinoma, 77 cases of tumors with clear cell changes such as malignant melanoma, clear cell renal cell carcinoma and alveolar soft part sarcoma, and 140 cases of other types of tumors such as ovarian endometrioid adenocarcinoma, pituitary tumor and thyroid papillary carcinoma) were studied.Immunohistochemical study for Arg-1 was performed on the paraffin-embedded tumor tissue.

RESULTSIn HCC, Arg-1 demonstrated both cytoplasmic and nuclear staining, with an overall sensitivity of 96.2% (75/78).In well, moderately and poorly differentiated HCC, the sensitivity was 15/15, 100% (41/41) and 86.4% (19/22), respectively. That was in contrast to negative staining for Arg-1 in all the 29 cases of metastatic tumors studied. The sensitivity, specificity, positive predictive value and negative predictive value of Arg-1 in distinguishing HCC from metastatic tumors was 96.2%, 100%, 100% and 90.6%, respectively. Cytoplasmic and membranous staining was observed in clear cell type of HCC. The overall sensitivity of Arg-1 expression in the 77 cases of tumors with clear cell changes was 14.3% (11/77), including 8/15 for malignant melanoma, 2/4 for ovarian clear cell carcinoma and 1/1 gall bladder adenocarcinoma with clear cell component.In malignant melanoma and ovarian clear cell carcinoma, only cytoplasmic staining was demonstrated. There was no expression of Arg-1 in the 140 cases of other tumor types studied.

CONCLUSIONSArg-1 is a sensitive and specific marker for HCC.It is a potentially useful immunohistochemical marker in distinguishing HCC from metastatic tumors. Though also expressed in malignant melanoma and ovarian clear cell carcinoma, Arg-1 shows a different staining pattern as compared with that in HCC.

Adenocarcinoma ; enzymology ; Adult ; Aged ; Arginase ; metabolism ; Carcinoma, Hepatocellular ; enzymology ; pathology ; secondary ; Cell Differentiation ; Diagnosis, Differential ; Female ; Gallbladder Neoplasms ; enzymology ; Humans ; Liver Neoplasms ; enzymology ; pathology ; secondary ; Male ; Melanoma ; enzymology ; Middle Aged ; Ovarian Neoplasms ; enzymology ; Stomach Neoplasms ; enzymology ; pathology

To investigate the expression of cyclooxygenase-2 (COX-2) in ovarian cancer cell lines, RT-PCR and immunocytochemistry were used to detect the expression of COX-2 in 5 ovarian cancer cell lines. The expression of COX-2 mRNA and protein was detected in all 5 cell lines. It is suggested that COX-2 is expressed in ovarian cancer cell lines, which provides a basis for the chemoprevention of ovarian cancer.
Cell Line, Tumor ; Cyclooxygenase 2/genetics ; Cyclooxygenase 2/*metabolism ; Ovarian Neoplasms/*enzymology ; Ovarian Neoplasms/*pathology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism

OBJECTIVETo study the relation between cyclooxygenase-2 (COX-2) protein expression and biologic behavior of ovarian carcinoma.

METHODSThe level of COX-2 protein expression was detected by Western Blot assay in 54 biopsy specimens from ovarian serous tumor patients and 10 normal ovarian samples.

RESULTSThe level of COX-2 protein expression and relative quantity in ovarian serous carcinoma (81.8%, 20.08 +/- 3.53) were statistically higher than those in the benign ovarian serous tumor (0, 15.04 +/- 0.12) and in the normal ovary (0, 15.33 +/- 0.60) (P < 0.05). The level of COX-2 protein expression and relative quantity in borderline ovarian serous tumor (90.0%, 20.61 +/- 3.03) were statistically higher than those in benign ovarian serous tumor and the normal ovary (P < 0.05). The level of COX-2 protein expression and relative quantity were not significantly different from ovarian serous carcinoma and borderline ovarian serous tumor (P > 0.05); as they were between the benign ovarian serous tumor and the normal ovary (P > 0.05). The level of COX-2 protein expression and relative quantity were not significantly different among different clinical stages (I + II and III + IV), different histological grades, with or without ascites or lymphatic metastasis either.

CONCLUSIONCOX-2 overexpression may be significantly related to the oncogenesis and development of ovarian serous carcinoma, which may be an early diagnostic parameter and, hence, an attractive target for chemopreventive strategy in the treatment of ovarian serous carcinoma.

Adult ; Aged ; Blotting, Western ; Cyclooxygenase 2 ; analysis ; genetics ; physiology ; Female ; Humans ; Middle Aged ; Ovarian Neoplasms ; enzymology ; etiology

BACKGROUNDSeprase plays an important role in malignant cell invasion and metastasis by degrading the extracellular matrix. However, its clinical significance remains largely unknown. The objective of the current study was to evaluate the expression of seprase in effusions from patients with epithelial ovarian carcinoma and its clinical values.

METHODSImmunohistochemistry was used to examine the expression of seprase protein in a series of 74 malignant peritoneal (n = 64) and pleural (n = 10) effusions from Norwegian patients with epithelial ovarian carcinoma. Additionally, 34 effusions were evaluated using the Western blotting. Nine reactive effusions, obtained from patients with benign lesions, served as a control group. Statistical analyses were carried out by Chi-square test and Kaplan-Meier method.

RESULTSIn the 74 malignant effusion specimens, 57 (77.02%) were positive for seprase, while only 2 (22.22%) of the control group were positively stained (P = 0.001). In the malignant effusions, 17 (22.97%), 22 (29.73%), 22 (29.73%), 13 (17.57%) had negative, weak, moderate and strong seprase protein expression, respectively. The expression of seprase protein was predominant in cytoplasm of carcinoma cells. Increased seprase protein was negatively associated with the overall survival rate of the patients (P = 0.03). However, there was no significant correlation between protein expression and FIGO stage, age, histology, and histological grade. By Western blotting, 27 of the 34 effusions showed the presence of both 170-kD dimeric form and 97-KD monomeric form of seprase while only 1 of the 34 had 170-KD dimeric form, which was consistent with the results of immunohistochemistry (P = 0.05).

CONCLUSIONSSeprase may be involved in the development of ovarian cancer, and is a potential predictive marker for the disease.

Ascitic Fluid ; enzymology ; pathology ; Blotting, Western ; Epithelial Cells ; pathology ; Female ; Gelatinases ; metabolism ; Humans ; Immunohistochemistry ; Membrane Proteins ; metabolism ; Middle Aged ; Neoplasm Staging ; Norway ; Ovarian Neoplasms ; enzymology ; pathology ; Pleural Effusion, Malignant ; enzymology ; pathology ; Serine Endopeptidases ; metabolism

OBJECTIVETo investigate expression and significance of hTERT, telomerase associated-regulation protein (TRAP) and proliferating cell nuclear antigen (PCNA) in ovarian epithelial tumors.

METHODS106 specimens of ovarian epithelial tumors and their clinical history were collected, including 54 cases of malignancy, 33 borderline cases and 19 benign tumor cases. Immunohistochemical staining for hTERT, TRAP and PCNA were performed. Follow-up information was obtained for 45 of 87 cases (malignancy in 54 and borderline malignancy in 33).

RESULTSThe expression of hTERT was significantly different between benign (4/19) and borderline (90.9%, 30/33) cases, benign and malignant (94.4%, 51/54) cases (P < 0.001), as was the expression of TRAP between benign (4/15) and malignant (77.8%, 28/36) cases (P < 0.001). The expression of hTERT and TRAP was not higher in stage III, IV ovarian cancer patients than in stage I and II (P > 0.05, P > 0.3). The expression of PCNA between benign (6.9 +/- 5.9)% and borderline (26.4 +/- 17.8)% cases, benign and malignant (51.8 +/- 22.1)% cases, and borderline and malignant cases were different, and were statistically significant (P < 0.01, P < 0.001, P < 0.05). 33 cases of borderline malignancy are all survive. In 54 cases of malignancy, 35 of them have metastasis (64.8%), including 5 cases of lymph nodes metastasis. 4 of them died (7.4%).

CONCLUSIONSThe expression of hTERT and TRAP is associated with the malignant degree of ovarian cancer, but does not correlate with stage. The expression of TRAP resembles hTERT, which may be a new tumor-associated gene. Telomerase activity is positively associated with PCNA.

DNA-Binding Proteins ; Female ; Humans ; Immunohistochemistry ; Neoplasm Staging ; Ovarian Neoplasms ; enzymology ; pathology ; Proliferating Cell Nuclear Antigen ; analysis ; Telomerase ; analysis ; Transcription Factors ; analysis

BACKGROUNDThe formation and growth of tumors are related to the synthesis of the DNA. The enzyme ribonucleotide reductase (RR) is an enzyme that regulates the total rate of DNA synthesis and thus plays a pivotal role in cell growth. Catalytic subunit M2 (RRM2) is the main unit modulating the ribonucleotide reductase enzymatic activity. This study aimed to investigate the expression of RRM2 mRNA and protein in patients with ovarian cancer and its relevance to diagnosis and clinical outcome of the patients.

METHODSRRM2 mRNA levels and protein expression were detected in 98 ovarian specimens with immunohistochemistry and real-time quantitative polymerase chain reaction (PCR). Expression of the RRM2 protein and correlation of the RRM2 gene expression with clinical pathological features were analyzed. The Kaplan-Meier test was used for evaluating RRM2 expression and time to progression and survival. The Cox proportional model was used to analyze the risk factors in prognosis of patients.

RESULTSPositive RRM2 immunostaining was found in 43 of 62 (69.4%) patients with epithelial ovarian cancer, 10 of 15 (66.7%) patients with borderline neoplasm, 4 of 15 (26.7%) patients with benign growths, and none of the normal group. The RRM2 mRNA levels were significantly over expressed in epithelial ovarian cancer (1.722 ± 0.639) and borderline ovarian neoplasms (1.365 ± 0.615), compared to the normal group (0.678 ± 0.446) and benign group (0.828 ± 0.545). Patients with ovarian caner in clinical FIGO-stages III-IV presented higher RRM2 gene expression than those in clinical FIGO-stages I-II. Furthermore, the survival of patients with low RRM2 mRNA level was significantly better than patients with high levels (P < 0.05). By Cox proportional risk model analysis, the risk of mortality of patients with high level expression of RRM2 mRNA was 2.553 times greater than those with low expression.

CONCLUSIONRRM2 expression closely correlates with the development of ovarian tumor and may serve as a novel predictive marker for diagnosis and prognosis of the disease.

Adolescent ; Adult ; Aged ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Ovarian Neoplasms ; enzymology ; genetics ; Real-Time Polymerase Chain Reaction ; Ribonucleoside Diphosphate Reductase ; genetics ; metabolism ; Young Adult

OBJECTIVETo investigate the signaling pathway through testing the effects of dexamethasone (Dex) on the activation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 kinase (p38) in HO-8910 cells.

METHODSActivation of the ERK1/2 and p38 was detected by Western blotting using the antibodies against the total ERK1/2 and p38 mitogen-activated protein kinases (MAPKs) protein and the phosphorylated forms of them.

RESULTSDex could suppress the activation of ERK1/2, while enhance the activation of p38 rapidly and strongly in a dose- and time- dependent manner. Neither effect could be blocked by RU486, the antagonist of glucocorticoid receptor (GR).

CONCLUSIONDex has rapid effects on the activation of ERK1/2 and p38, and these effects are not mediated by GR.

Cell Division ; physiology ; Dexamethasone ; pharmacology ; Enzyme Activation ; Female ; Glucocorticoids ; pharmacology ; Humans ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; metabolism ; Ovarian Neoplasms ; enzymology ; pathology ; Signal Transduction ; Tumor Cells, Cultured ; p38 Mitogen-Activated Protein Kinases

OBJECTIVETo investigate the heparitinase (HPA) expression and cell invasiveness in human ovarian cancer cell line SKOV3 during hypoxia, and explore their relationship with hypoxia inducible factor-1alpha (HIF-1alpha).

METHODSSKOV3 cells were incubated with normoxia, hypoxia, and hypoxia plus rapamycin, respectively. SKOV3 cells of hypoxia group were incubated in 5% CO2 + 1% O2. Cells in hypoxia plus rapamycin group were incubated with 10 ng/ml of rapamycin before cultured under hypoxic condition. Cells in each group were collected for analysis after incubated with hypoxia for 12, 24, and 36 hours, respectively. Western blotting and RT-PCR were performed to detect the expressions of HIF-1alpha and HPA. Cell invasiveness was measured by matrigel invasion assay.

RESULTSWestern blotting showed that the expression of HIF-1alpha significantly increased compared with normoxic group (P < 0.05). However, hypoxia had no obvious impact on HIF-1alpha mRNA expression. The expressions of HPA protein and mRNA of SKOV3 cells of hypoxia group were significantly higher than normoxic group (P < 0.05). The up-regulation of HPA expression in hypoxic group decreased after the utilization of rapamycin. The cell invasion of hypoxic group was significantly higher than that of normoxic group (P < 0.05); meanwhile, the expression of HPA was positively correlated with the invasiveness of SKOV3 cells (r = 0.9863, P < 0.05).

CONCLUSIONHypoxia may promote the expression of HPA and the invasiveness of SKOV3 cells through the HIF-1alpha pathway, which plays an important role in the pathogenesis of ovarian cancer.

Cell Hypoxia ; Cell Line, Tumor ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Neoplasm Invasiveness ; Ovarian Neoplasms ; enzymology ; genetics ; metabolism ; pathology ; Polysaccharide-Lyases ; genetics ; metabolism ; Signal Transduction ; Up-Regulation

OBJECTIVETo study the anti-tumor immunotherapeutic effect induced by the suicidalcancer vaccine FC/TK, and to evaluate the safety of this vaccine.

METHODSThe suicidal cancer vaccine, named FC/TK, was prepared by fusion of suicide gene (HSVI,-TK gene) -modified ovarian carcinoma NuTu-19 cells with rat bone marrow-derived dendritic cells (DCs). The morphology of FC/TK was evaluated by scanning electron microscopy. The stimulatory effect of FC/TK on T cells was determined by T cell proliferation assay. In immunotherapeutic studies in vivo, Fischer344 rats were injected subcutaneously with NuTu-19 cells, followed by treatment of FC/TK on days 7 and 14, compared to controls treated with irradiated FC/TK, FC or PBS, respectively. Tumor incidence and volume were measured in 90 days after challenge. To determine the killing effect of FC/TK in vivo, TUNEL assays were applied to detect apoptotic cell death in spleen of vaccinated rats with prodrug ganciclovir administration.

RESULTSFC/TK cells were of irregular shape with surface membrane processes. Compared to the control groups, FC/TK significantly promoted T cell proliferation (P <0.01). The rats vaccinated with FC/TK and FC significantly inhibited the tumor growth compared to rats vaccinated with irradiated FC/TK (P <0.05) or with PBS ( P <0.01). The immunotherapeutic effect induced by FC/TK was similar to that using FC. Fluorescence microscopy showed that fluorescein-stained FC/TK cells migrated into spleen also showed to be TUNEL-positive, suggesting that the FC/TK cells were killed by ganciclovir in vivo.

CONCLUSIONOur data indicate that suicidal cancer vaccine is an effective and safe therapy for ovarian carcinoma and may serve as a broadly applicable approach for other cancer vaccines in the future.

Animals ; Apoptosis ; drug effects ; Cancer Vaccines ; immunology ; Cell Fusion ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dendritic Cells ; cytology ; immunology ; Female ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; Herpesvirus 1, Human ; enzymology ; genetics ; Immunotherapy ; methods ; Microscopy, Electron, Scanning ; Microscopy, Fluorescence ; Neoplasms, Experimental ; enzymology ; pathology ; therapy ; Ovarian Neoplasms ; enzymology ; pathology ; therapy ; Rats ; Rats, Inbred F344 ; Survival Analysis ; T-Lymphocytes ; drug effects ; metabolism ; pathology ; Thymidine Kinase ; genetics ; metabolism ; Transfection

OBJECTIVETo investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells.

METHODSCisplatin-induced apoptosis were stained with DAPI and was assessed microscopically in human epithelial adenocarcinoma ovarian cell line SKOV3 cells. ERK activation was determined by Western blotting using an anti-phospho-ERK antibody to detect ERK activity. The effect of PD98059 on ERK activity induced by cisplatin was detected by MTT assay.

RESULTSMarked apoptosis of SKOV3 cells resulted from 48 hours treatment with 20 microg/mL cisplatin. Strong activation of ERK was led to by 15 microg/mL cisplatin. Dose response and time course of cisplatin induced apoptosis in SKOV3 cells. Cisplatin-induced ERK activation occurred at 12 hours and increased to highest induction at 24 hours by Western blotting. The effect of PD 98059 on ERK activity induced by cisplatin at the concentration of 100 micromol/L PD 98059. Statistically significant decreased in cell survival were observed with 100 micromol/L PD 98059 at 15 and 20 microg/mL cisplatin (P < 0.05).

CONCLUSIONSCisplatin activates the ERK signaling pathway in ovarian cancer cell line SKOV3. Inhibition of ERK activity enhances sensitivity to cisplatin cytotoxity in ovarian cancer cell line SKOV3. Evaluation of ERK activity could be useful in predicting which ovarian cancer will response most favorably to cisplatin therapy.

Adenocarcinoma ; enzymology ; pathology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Enzyme Activation ; drug effects ; Female ; Flavonoids ; pharmacology ; Humans ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; metabolism ; Ovarian Neoplasms ; enzymology ; pathology ; Signal Transduction