OBJECTIVETo explore the role of Ca(2+) in nanosecond steep pulse (NSP)-induced apoptosis of human ovarian carcinoma cell line SKOV3 in vitro.

METHODSThe early apoptotic rate of SKOV3 cells treated with NSP was detected by Annexin V/PI double staining and flow cytometry. MTT assay was used to detect the viability of the cells pretreated with BAPTA-AM (0, 25, 50 and 100 µmol/L) chelation for 1 h to increase the intracellular free Ca(2+) prior to NSP exposure, and the cell morphological changes and caspase 12 expression were detected using Hoechst 33342 staining and Western blotting, respectively.

RESULTSFlow cytometry showed that NSP induced early apoptosis of SKOV3 cells, and the optimal effect was achieved with the treatment parameter configuration of field strength of 90 kV/cm, pulse width of 100 ns, frequency of 1 Hz, and exposure time of 30 s. The highest early apoptotic rate and necrosis rate was (60.31∓5.67)% and (1.35∓0.39)%, respectively. Pretreatment with BAPTA-AM chelation prior to NSP exposure significantly increased the cell viability (P<0.05), and resulted also in lowered apoptosis rate and decreased expression of caspase 12 (P<0.05).

CONCLUSIONNSP can induce apoptosis in SKOV3 cells. Increased intracellular free Ca(2+) functions as an important mediator in NSP-induced cell apoptosis, which may also involve Ca(2+)-mediated endo- plasmic reticulum pathway.

Apoptosis ; Calcium ; chemistry ; pharmacology ; Cell Line, Tumor ; Female ; Humans ; Nanoparticles ; Ovarian Neoplasms ; metabolism ; pathology

Mucinous neoplasms occur rarely in association with cystic teratoma, Sertoli-Leydig cell tumor, granulosa cell tumor or carcinoid tumor. Several cases of an ovarian stromal tumor with minor sex-cord elements have been reported in the literatures. However, there has been no report about an ovarian mucinous neoplasm coexisting with a stromal tumor with sex-cord elements yet. We report a case of an ovarian neoplasm composed of both mucinous cystadenoma and stromal tumor with minor sex-cord elements in a 58-yr-old female. The ovary including the mass measured 5 cm in size. On section, it revealed an unilocular cyst (4.5 cm in diameter) filled with mucinous fluid. There was a round, yellow, solid nodule, 1.5 cm in diameter within the wall. Microscopically, the cyst was lined by a single layer of endocervical mucinous epithelium and the nodule was composed of spindle cells showing an intersecting and whorled arrangement. There were cell nests showing polygonal shape with abundant cytoplasm among the spindle cells. They showed immunoreactivity for inhibin and did not have any connection with the adjacent mucinous epithelium. Therefore, we interpret the mucinous cystadenoma as having arisen de novo.
Cystadenoma, Mucinous/chemistry/*pathology ; Female ; Human ; Inhibin/analysis ; Middle Age ; Ovarian Neoplasms/chemistry/*pathology ; Sertoli-Leydig Cell Tumor/pathology ; Stromal Cells/*pathology

OBJECTIVETo determine the apoptotic and proliferative activities in various ovarian epithelial tumors.

METHODSFormalin-fixed, paraffin-embedded tissues of 86 ovarian epithelial tumors, including 52 adenocarcinomas, 23 borderline tumors and 11 cystadenoma, were retrieved. Apoptotic (AI) and proliferative (PI) index were estimated using the monoclonal antibodies: M30, Ki-67 and Ki-S1 in these tumors. Quantitative assessment of AI and PI was estimated by calculating the percentage of positive cells among no less than 1000 tumor cells.

RESULTSStatistically significant difference in AI was found between benign and borderline tumors or carcinomas (P = 0.028, 0.001, respectively). Significant differences in PI, as assessed by both Ki-67 and topo IIalpha, were demonstrated between carcinomas and benign or borderline tumors (both P < 0.001). Benign tumors had both low PI and AI; borderline tumors had lower PI but higher AI, while adenocarcinomas had both high proliferative and high apoptotic rates. Among borderline tumors, serious tumors had significantly lower AI and higher PI than mucinous ones.

CONCLUSIONThe results suggest that apoptotic and proliferative activities play important roles in the pathogenesis and development of ovarian borderline and malignant tumors. The high apoptotic rate in borderline tumor may explain its relatively indolent behavior while the high proliferative rate in carcinomas tends to explain its aggressive behavior.

Antigens, Neoplasm ; Apoptosis ; Carcinoma, Endometrioid ; chemistry ; pathology ; Cell Division ; Cystadenocarcinoma, Serous ; chemistry ; pathology ; Cystadenoma, Mucinous ; chemistry ; pathology ; Cystadenoma, Serous ; chemistry ; pathology ; DNA Topoisomerases, Type II ; analysis ; DNA-Binding Proteins ; Female ; Humans ; Ki-67 Antigen ; analysis ; Ovarian Neoplasms ; chemistry ; pathology

OBJECTIVETo observe the protective effects of follicle stimulating hormone (FSH) on human epithelial ovarian cancer cell apoptosis induced by cisplatin (DDP).

METHODSOVCAR3-FSHR cell were treated with DDP and FSH at serials of concentrations, MTT assay was used to examine the growth inhibition of OVCAR3-FSHR cell after treatment with DDP and FSH. Flow cytometry was used to analyze the change of cell cycle and percentage of apoptosis.

RESULTSIt was revealed that FSH decreased the growth inhibition induced by DDP. We also demonstrated that FSH reduced the S-phase percentage compared with the DDP only groups after treatment for 24 hours and reduced apoptosis percentage after 48 hours treatment with DDP.

CONCLUSIONIt is suggested that FSH can protect the apoptosis induced by DDP. It also suggests that FSH may be an important chemoresistent reason for the chemotherapy of ovarian cancer.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cisplatin ; pharmacology ; Female ; Flow Cytometry ; Follicle Stimulating Hormone ; pharmacology ; Humans ; Ovarian Neoplasms ; chemistry ; pathology ; Receptors, FSH ; analysis ; Tumor Cells, Cultured

OBJECTIVETo analyze the clinicopathological features of ovarian tumors with endocrine function.

METHODSTwenty-four cases of ovarian tumor with endocrine manifestation were collected from the hospital. Their clinical presentation and histopathologic features were reviewed, along with a panel immunohistochemistry stainings (EnVision method). The antibodies were AE1/AE3, epithelial membrane antibody (EMA), alpha-inhibin, calretini and smooth myoglobin (SMA).

RESULTSThe main clinical endocrinological manifestations were related to an excess production of sex steroids. Histologically, the principle histological subtype of these tumors was ovarian sex cord-stroma tumors, including 13 cases ovarian type (8 granulosa cell tumors, 2 thecofibromas, 3 sclerosing stromal tumors), 7 cases testicular type (1 sertoli cell tumors, 5 sertoli-Leydig cell tumors, 1 Leydig cell tumor, and 2 cases of steroid cell tumor (NOS). Another 2 cases were ovarian epithelial tumors. Grossly, 50% (11/22) ovarian sex cord-stromal tumors were less than 5 cm in diameter. However, 4 tumors were quite larger, up to 18 cm in diameter. Most of these tumors were solid or solid-cystic and their cut surfaces were brown, pink, yellow or grey in color. The 2 primary ovarian epithelial tumors were larger, being 12 cm and 14 cm in diameter, respectively. Immunohistochemically, ovarian sex cord-stromal tumors showed positive staining for alpha-inhibin in all cases (22/22) and for calretinin in majority cases (18/22), and that the intensity of reactivity correlated with the degree of tumor differentiation. The non-neoplastic, luteinized stromal cells in 2 ovarian primary epithelial tumors also showed positive staining. Five cases of fibrothecomas and sclerosing stroma tumors were all positive for SMA. Although 6 of the 22 ovarian sex cord-stromal tumors were AE1/AE3 positive, all were EMA negative.

CONCLUSIONSMost endocrinological syndromes in ovarian neoplasia reflect an overproduction of sex steroids, but the clinical manifestations do not correlate with the tumor histological subtypes. Most functional ovarian tumors are sex cord-stromal tumors and are usually of small to medium in size, but few are larger or giant. The size of the tumor does not correlate with the duration and the degree of clinical manifestations. Ovarian tumors of non-sex cord-stromal type may also be clinically functional. The immunohistochemical results suggests that alpha-inhibin and/or calretinin expression are useful markers in support of a diagnosis of sex cord-stromal tumor of the ovary. Although some of these tumors are AE1/AE3 positive, EMA negativity may be useful for the differential diagnosis with epithelial ovarian tumors.

Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; analysis ; Calbindin 2 ; Female ; Gonadal Steroid Hormones ; secretion ; Granulosa Cell Tumor ; chemistry ; pathology ; secretion ; Humans ; Immunohistochemistry ; Inhibins ; analysis ; Middle Aged ; Ovarian Neoplasms ; chemistry ; pathology ; secretion ; S100 Calcium Binding Protein G ; analysis ; Sertoli-Leydig Cell Tumor ; chemistry ; pathology ; secretion ; Sex Cord-Gonadal Stromal Tumors ; chemistry ; pathology ; secretion

Coexpression of Kit ligand and c-kit has been reported in some gynecologic tumors. To determine whether imatinib mesylate is useful in ovarian epithelial tumors, we performed immunohistochemical and mutational analysis. The cases consisted of 33 cases, which included 13 serous cystadenocarcinomas, 1 borderline serous tumor, 8 mucinous cystadenocarcinomas, 6 borderline mucinous tumors and 5 clear cell carcinomas. Five cases of serous cystadenoma and 5 cases of mucinous cystadenoma were also included. In the immunohistochemical study, 3 cases (3/6, 50%) of borderline mucinous cystic tumor and two cases (2/8, 25%) of mucinous cystadenocarcinoma show positive staining for KIT protein. Only one case (1/13, 7.7%) of serous cystadenocarcinoma had positive staining. On mutational analysis, no mutation was identified at exon 11. However, two cases of borderline mucinous tumors and one case of mucinous cystadenocarcinoma had mutations at exon 17. In these cases, the immunohistochemistry also shows focal positive staining at epithelial component. Although, KIT protein expression showed higher incidence in mucinous tumors than serous tumors, they lack KIT-activating mutations in exon 11. Thus, ovarian surface epithelial tumors are unlikely to respond to imatinib mesylate.
Adult ; Aged ; Cystadenocarcinoma, Mucinous/genetics/metabolism/pathology ; Cystadenoma, Mucinous/genetics/metabolism/pathology ; Cystadenoma, Serous/genetics/metabolism/pathology ; DNA Mutational Analysis ; DNA, Neoplasm/chemistry/genetics ; Epithelial Cells/chemistry/metabolism/pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Middle Aged ; Mutation ; Ovarian Neoplasms/genetics/metabolism/*pathology ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Proto-Oncogene Proteins c-kit/biosynthesis/*genetics

OBJECTIVETo investigate the effects of PsL5F (ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid, an extract from Pteris semipinnata L) on the expression of nuclear receptor subfamily 1, group D, member 1 (Nr1d1) in highly metastatic ovarian carcinoma HO-8910PM cells, and its mechanisms.

METHODMicroarray Chip was used to examine the level of Nr1d1 mRNA expression on HO-8910PM cells treated with PsL5F. Fluorescent quantitative real-time PCR assay and Western blot were performed to verify the effects of PsL5F on Nr1d1 mRNA and protein expression.

RESULTAfter 24 h treatment of 100 micromol x L(-1) PsL5F, the mRNA and protein levels of Nr1d1 in HO-8910PM cells were 35.34 +/- 1.07 and 7.71 +/- 0.43 times compared to those of control group (P < 0.01, P < 0.01), respectively.

CONCLUSIONPsL5F can up-regulate significantly the expression of Nr1d1 in HO-8910PM cells. Antitumor effects and its mechanisms of PsL5F in HO-8910PM cells may be involved in the up-regulation of Nr1d1 expression.

Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Female ; Gene Expression ; drug effects ; Humans ; Neoplasm Invasiveness ; Ovarian Neoplasms ; pathology ; Piperidones ; pharmacology ; Pteris ; chemistry ; RNA, Messenger ; drug effects ; metabolism

To prepare polyrotaxane-camptothecin conjugates and evaluate its anti-tumor effect, polyrotaxane-camptothecin conjugates were successfully synthesized, and the release behavior was performed; MTT assay and cell morphology were used to examine the inhibition of cells' proliferation effect in vitro. The experimental study of the antitumor effect on S180 mice in vivo was also performed to further evaluate the anti-tumor effect of conjugate. The result showed polyrotaxane-camptothecin conjugates can effectively inhibit the proliferation in a dose dependent effect. In vivo study and cell morphology observation of S180 mice showed significant decrease in growth of tumor, degree of tumor infiltration and blood vessel number. The result indicated anti-tumor mechanism may be through affect the angiogenesis and reduced blood supply to tumor cells and then leading to necrosis.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Camptothecin ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclodextrins ; chemistry ; Drug Carriers ; Drug Compounding ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Ovarian Neoplasms ; pathology ; Poloxamer ; chemistry ; Rotaxanes ; chemistry ; Sarcoma 180 ; pathology ; Tumor Burden ; drug effects

Drug delivery system (DDS) is a novel approach to overcome multidrug resistance (MDR) in tumors nowadays. This work was designed to investigate a new micellar delivery system for in vitro reversal of resistant ovarian tumor cells, based on a nonionic triblock copolymer Pluronic P105 and paclitaxel (PTX). The PTX-loaded polymeric micelles (P105/PTX) were prepared by thin film-hydration methods. Based on the results of single factor experiments, the P105/PTX micelle formulation was optimized by employing the central composite design-response surface methodology. The physico-chemical properties of the P105/PTX micelles were characterized, including micelle size, drug loading coefficient, in vitro release behavior, etc. The cytotoxicity of the P105/PTX micelles was assessed against human ovarian tumor cell line, SKOV-3/PTX, by a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl (MTT) assay. In order to understand the possible mechanism of Pluronic effects in resistant tumor cells, cellular uptake study of micellar PTX or Rhodamine-123 (R-123) was also carried out. The results showed that the micelle size was about 24 nm with drug loading coefficient of 1.1% and PTX concentration of 700 microg x mL(-1). The cumulative release amount of PTX from the P105/PTX micelles was only 45.4% in 6 h (P < 0.05) and 79.6% in 24 h, whereas Taxol injection in 6 h released 95.2% PTX. The IC50 values of the P105/PTX micelles and Taxol injection against SKOV-3/PTX were 1.14 and 5.11 microg x mL(-1), and resistance reversion index (RRI) was 9.65 and 2.15, respectively. The micellar PTX or R-123 exhibited a significant increase in cellular uptake in resistant SKOV-3/PTX cells compared with free PTX or R-123. These results indicated that PTX could effectively be solubilized by Pluronic P105 block copolymers via thin film-hydration process and formulation optimization, producing nano-scale polymeric micelles with sustained release property in vitro. The P105/PTX micelles were effectively able to reverse resistance to PTX in SKOV-3/PTX tumor cells compared with Taxol injection or free PTX solution, and the enhanced cytotoxicity in the resistant SKOV-3/PTX cell was related to the improved cellular uptake of PTX by Pluronic P105 copolymers.
Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; pharmacology ; Cell Line, Tumor ; Drug Carriers ; Drug Delivery Systems ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Excipients ; chemistry ; Female ; Humans ; Inhibitory Concentration 50 ; Micelles ; Ovarian Neoplasms ; metabolism ; pathology ; Paclitaxel ; administration & dosage ; chemistry ; metabolism ; pharmacology ; Particle Size ; Poloxamer ; chemistry

OBJECTIVETo evaluate the effect of Group A streptococcus preparation (sapylin) combined with cisplatin on the malignant peritoneal effusion in patients with advanced cancer.

METHODSSixty advanced cancer patients with large amount of peritoneal effusion were divided into two groups: sapylin + cisplatin (DDP) group (30 patients) and control group (DDP alone, 30 cases). All the chemotherapeutic agents were injected intraperitoneally through a catheter.

RESULTSIn sapylin + cisplatin(DDP) group, 11 (36.7%) patients showed CR and 16 (53.3%) PR. The overall response rate (CR + PR) was 90.0%. Those of DDP alone group were 16.7% and 46.7%, the overall response rate was 63.3%. The main adverse effects were fever, nausea, and vomiting.

CONCLUSIONCombined Group A streptococcus preparation (sapylin) and cisplatin is more effective than cisplatin alone for the malignant peritoneal effusion in patients with advanced cancer, and the adverse effects are tolerable.

Adjuvants, Immunologic ; administration & dosage ; Adult ; Aged ; Antineoplastic Agents ; administration & dosage ; Ascitic Fluid ; pathology ; Bacterial Proteins ; administration & dosage ; Biological Products ; administration & dosage ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Female ; Humans ; Immunologic Factors ; administration & dosage ; Injections, Intraperitoneal ; Male ; Middle Aged ; Ovarian Neoplasms ; pathology ; therapy ; Peritoneal Neoplasms ; secondary ; therapy ; Stomach Neoplasms ; pathology ; therapy ; Streptococcus pyogenes ; chemistry