Weakened spindle checkpoint with reduced BubR1 expression in paclitaxel-resistant ovarian carcinoma cell line SKOV3-TR30.

Objective: Paclitaxel sensitivity has recently been associated with the spindle checkpoint. The aim of our study is to investigate the status of spindle checkpoint and the alteration of its major components in phenotype with acquired paclitaxel resistance in ovarian carcinoma.

Methods: A paclitaxel-resistant ovarian carcinoma cell line, SKOV3-TR30, with the resistant ability of 27-fold greater than its parental cell line, was derived from SKOV3 cell line.

[Deletion of OPCML gene and promoter methylation in ovarian epithelial carcinoma].

Objective: To investigate the expression of OPCML gene in ovarian epithelial carcinoma and determine the relationship between mRNA expression and methylation of their promoters.

Method: Twenty normal ovarian tissues and 89 ovarian epithelial tumor specimens (72 malignant, 17 benign), as well as 3 ovarian carcinoma cell lines (SKOV-3, CAOV3, and 3AO), were collected for detection of OPCML gene expression by reverse transcription-polymerase chain reaction and for detection of promoter methylation by restriction enzyme cut analysis from 7. 1999 to 7.

Proportion of CD4+CD25+ regulatory T cell is increased in the patients with ovarian carcinoma.

Objective: To study the frequency of the CD4+CD25+ regulatory T cells (Tregs) in the patients with ovarian carcinoma and its possible mechanism.

Methods: The percentages of CD4+CD25+ Tregs in the peripheral blood lymphocytes (PBLs), tumor infiltrating lymphocytes (TILs) and tumor associated lymphocytes (TALs) from 13 patients with ovarian carcinoma and in the PBLs from 14 healthy women were determined by flow cytometry. The expression of CD69 on CD4+PBLs from the patients was detected.

Ovarian carcinoma cells effectively inhibit differentiation and maturation of dendritic cells derived from hematopoietic progenitor cells in vitro.

Loco-regional dissemination of ovarian carcinoma is associated with immunosuppression of the peritoneal cavity. One marked characteristic of the peritoneal immunity in this disease is the defective function of dendritic cells (DCs). In this study, the affect of ovarian carcinoma cells on DCs derived from hematopoetic progenitor cells was observed.

[Tumor-infiltrating dendritic cells in epithelial ovarian carcinoma and correlation with the expression of vascular endothelial growth factor].

Objective: To investigate the density and activation status of tumor infiltrating dendritic cells (TIDC) in epithelial ovarian carcinoma (EOC) and correlation with the expression of vascular endothelial growth factor (VEGF).

Methods: Streptavidin-peroxidase (SP) and Picture two-step immunohistochemistry methods were used to detect S-100(+), CD(83)(+) TIDC and the expression of VEGF in 57 primary EOCs, 32 benign ovarian tumors (benign control) and 16 normal ovarian tissues (normal control).

Results: (1) Two types of heterogeneous distribution pattern of TIDC in EOC were observed under the microscope.

[Possibility of PTEN expression on predicting pathologic risk factors of endometrioid adenocarcinoma before operation].

Background & Objective: Fully estimating pathologic risk factors is important for selecting operation and predicting prognosis for endometrioid adenocarcinoma. Phosphatase and tension homology deleted on chromosome ten (PTEN), taken as the housekeeping gene of endometrium, has the highest mutation rate in endometrioid adenocarcinoma. This study was to investigate the effect of PTEN on predicting pathologic risk factors of endometrioid adenocarcinoma before operation.

PTEN promoter methylation and protein expression in normal early placentas and hydatidiform moles.

Objective: To investigate the relationship between PTEN promoter methylation and protein expression, and the possible involvement of the PTEN gene in development of gestational trophoblasts and the pathogenesis of hydatidiform moles.

Methods: DNA was extracted from choria of normal early placentas, partial hydatidiform moles, complete hydatidiform moles, and invasive moles, and overdigested by methylation-sensitive endonuclease HpaII. The PTEN promoter was amplificated by polymerase chain reaction.

[Expressions of VEGF/VEGFRs and activation of STATs in ovarian carcinoma].

Objective: To study the expressions of VEGF/VEGFRs and activation of STATs in ovarian epithelial carcinoma, and to elucidate direct effect of VEGF on ovarian carcinoma cells.

Methods: Tissue samples from 42 women with primary ovarian epithelial carcinoma (OVCA), 29 with begnin ovarian tumor (OVBT) and 11 with normal ovarian tissue (NOV) were collected. LSAB immunohistochemical staining was used to determine the expression of VEGF, VEGFR1, VEGFR2 and activated STATS (P-STAT1, P-STAT3, P-STAT5, P-STAT6) proteins.

[Effects of vascular endothelial growth factor on differentiation and function of dendritic cells generated from CD34+ hematopoietic progenitor cells in vitro].

Objective: To investigate the effects of vascular endothelial growth factor (VEGF) on differentiation and function of dendritic cells derived from CD34+ hematopoietic progenitor cells.

Methods: After isolation from umbilical cord blood with a high-gradient magnetic cell sorting system (MACS), the CD34+ cells were cultured with a cocktail cytokines for differentiating into dendritic cells (DC). The cells were stimulated by VEGF (25 ng/ml) either at the beginning or at day 9 of culture.

[Development of HPV16 positive cervical cancer model in the hu-PBL-SCID mouse and its immunological features].

Objective: To develop a HPV16 positive cervical cancer model in the hu-PBL-SCID mouse and investigate its immunological features.

Methods: Thirty-two CB17SCID mice were randomly divided into 4 groups: group A (5 mice) subcutaneously injected with phosphate-buffered saline, group B (5 mice) intraperitoneally injected with human peripheral blood lymphocyte (PBL) for immune reconstruction, group C (11 mice) subcutaneously injected with human cervical carcinoma cell line SiHa, and group D (11 mice) intraperitoneally injected with PBL and subcutaneously injected with SiHa cells after 24 hours of PBL transplantation. The tumor growth, behaviors and status of xenogeneic graft versus host disease (XGVHD) were observed.

VEGF, VEGFRs expressions and activated STATs in ovarian epithelial carcinoma.

Objective: To investigate the expressions of vascular endothelial growth factor (VEGF), VEGFRs and activation of signal transducers and activators of transcription (STATs) in ovarian epithelial carcinoma and the relationships among them.

Methods: The tissue samples of 42 primary ovarian epithelial carcinoma, 29 benign ovarian tumor and 11 normal ovarian tissue were used to determine the expression of VEGF, VEGFR1, VEGFR2, P-STAT1, P-STAT3, P-STAT5 and P-STAT6 proteins by immunohistochemical staining.

Results: VEGF in ovarian carcinomas was significantly higher than that in benign and normal ovarian tissues.

Mismatch repair gene promoter methylation and expression in hydatidiform moles.

Methods: In this study, to investigate the significance of mismatch repair genes (MMR) promoter methylation and expression in the pathogenesis and malignant transformation of hydatidiform moles, we assayed promoter methylation and protein expression of the MMR genes hMLH1 and hMSH2 in gestational trophoblastic diseases (GTDs). DNA was extracted from normal placentas, partial hydatidiform moles, complete hydatidiform moles, and invasive moles, over-digested by methylation-sensitive endonuclease Hpa II, and then the promoters were amplificated by polymerase chain reaction. The protein expression was detected by immunohistochemistry.

[Phosphorylation of signal transducer and activators of transcription 3 induced by vascular endothelial growth factor in ovarian carcinoma cell line in vitro].

Objective: To observe phosphorylation of signal transducer and activators of transcription 3 (STAT3) in Caov-3 induced by vascular endothelial growth factor (VEGF), and to investigate molecular mechanisms of the effect of VEGF on ovarian carcinoma cells.

Methods: The expressions of phosphorylated STAT3 in Caov-3 induced by VEGF were detected by immunocytochemistry and Western blot methods. Furthermore, the relationship between STAT3 phosphorylation and VEGF stimulation in ovarian carcinoma cells was investigated using a peptide which could specifically bind VEGFR2 and thus block the binding of VEGF to its receptors.

Vascular endothelial growth factor (VEGF) and ovarian carcinoma cell supernatant activate signal transducers and activators of transcription (STATs) via VEGF receptor-2 (KDR) in human hemopoietic progenitor cells.

Objective: To investigate the STATs signaling pathway activated by VEGF in human hemopoietic progenitor cells.

Methods: CD34(+) hemopoietic progenitor cells, which isolated from umbilical cord blood, were treated with VEGF or culture supernatant of ovarian carcinoma cell line which could secrete large amount of VEGF, phosphorylation and nuclear translocation of STAT3 and STAT5 were then detected by Western Blot and immunocytochemistry. Expression of VEGFR2/KDR on CD34(+) cells was studied by immunocytochemistry.

[Relationship between interleukin-10 level in ascites of patients with primary ovarian epithelial carcinoma and immune defect in peritoneal cavity].

Background & Objective: As a multifunctional Th2-cytokine, interleukin-10 (IL-10)plays a major role in the immune response. It is well known that IL-10 is an immunosuppressive cytokine, and participates in the development and progression of various tumors. In this study, we investigated the relationship between the IL-10 level in the ascites of the patients with primary ovarian epithelial carcinoma (POEC) and immune defect in the peritoneal cavity.

[Analysis of the risk factors for ovarian metastasis in patients with endometrial carcinoma].

Objective: To study the risk factors for ovarian metastasis in patients with endometrial carcinoma.

Method: The pathological and clinical features and outcomes of endometrial carcinoma patients who were diagnosed and treated in our hospital from Jan 1996 to Dec 2002 were retrospectively reviewed and analyzed.

Results: Of the 321 cases reviewed, 15 (4.

[Role of three-dimensional transvaginal sonography in evaluation of the depth of myometrial invasion of endometrial carcinoma].

Objective: To investigate the value of three-dimensional transvaginal sonography (3-DTVS) in diagnosing depth of myometrial invasion (MI) and analyze factors that may influence 3-DTVS diagnosis.

Methods: Fifty-three patients with endometrial carcinoma proven by histological diagnosis postoperatively in Women's Hospital, Zhejiang University from 2002 to 2003 were included in the study. All patients underwent primary surgery and 2-DTVS and 3-DTVS examinations within 7 days before operation.

[Activation of signal transducers and activators of transcription induced by vascular endothelial growth factor in CD34+ hematopoietic progenitor cells in vitro].

Objective: To investigate the activation pattern of signal transducers and activators of transcription (STAT) induced by vascular endothelial growth factor (VEGF) in CD34+ hematopoietic progenitor cells, and gain an insight into the molecular mechanism and signal transduction pathway of VEGF that has an effect on CD34+ hematopoietic progenitor cells.

Methods: After isolated from umbilical cord blood by using a high-gradient magnetically activated cell sorting system (MACS), CD34+ cells were stimulated by VEGF (50 ng/ml) for different time (0, 15, 30, 45, 60, 90 min) to detect the tyrosine phosphorylation and nuclear translocation of STAT-3 and STAT-5 with Western blot and immunocytochemistry methods. The expression of VEGF receptor-2 (VEGFR2) on the membrane of CD34+ progenitor cells was examined by immunocytochemistry.

Effect of interleukin-7 gene transfection into ovarian carcinoma cell line SKOV3 in vitro and in vivo.

Objective: To investigate the effect of interleukin-7 (IL-7) gene transfection into an established ovarian carcinoma cell line (SKOV3) in vitro and evaluate the tumorigenicity of SKOV3-IL-7 in severe combined immunodeficient (SCID) mice.

Methods: IL-7 gene was transfected into SKOV3 cells by liposome. IL-7 mRNA and protein of SKOV3-IL-7 and their parental control cells were detected by reverse transcriptive-polymerase chain reaction (RT-PCR) and Western blot, respectively.

Ovarian carcinoma cells inhibit T cell proliferation: suppression of IL-2 receptor beta and gamma expression and their JAK-STAT signaling pathway.

Deficient T cell immune function and intracellular signaling in cancer patients may result from effects of tumors or their products on lymphocytes. Recently, it was demonstrated that several ovarian carcinoma cell lines could produce soluble factors that inhibited T cell proliferation. The aim of this study is to assess the effect of supernatants from 3 ovarian carcinoma cell lines (OVCAR3, CAOV3, SKOV3) on signal transduction elements that are linked to the IL-2R and its JAK-STAT pathway.

[Related-factor analysis on efficacy of re-platinum chemotherapy in platinum-sensitive recurrent ovarian carcinoma].

Objective: To analysis the related factors of efficacy of re-platinum chemotherapy in platinum-sensitive recurrent ovarian carcinoma.

Methods: Forty-one patients with platinum-sensitive recurrent ovarian cancer from Jan. 1998 to Oct.

[Experimental study on apoptosis in T cells induced by ovarian carcinoma cells].

Objective: To investigate apoptosis in T cells induced by ovarian carcinoma cells and analyze the role of nitric oxide and intracellular free calcium in this process.

Methods: Apoptosis induced by ovarian carcinoma cell lines supernatants was investigated by electron microscopy and flow cytometry. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of inducible nitric oxide synthase (iNOS) mRNA.

[Biologic characteristics of intraperitoneal transplantation model of human ovarian carcinoma in severe combined immunodeficiency mice].

Objective: To develop a human ovarian carcinoma SKOV3 model in severe combined immunodeficiency (SCID) mouse and to study its biologic characteristics.

Methods: Human ovarian carcinoma SKOV3 cells were injected intraperitoneally into female SCID mouse to establish a transplantation model of human ovarian carcinoma. The biological characteristics, metastasis and morphology of transplanted tumors were studied.

[Hypermethylation of hMLH1 and microsatellite instability in ovarian mucinous tumors].

Objectives: To investigate the role of hMLH1 promoter hypermethylation and microsatellite instability (MSI) in the development of ovarian mucinous tumors.

Methods: One hundred and seven of paraffin-embedded specimens of ovarian mucinous tumors (malignant 49, borderline 35, and benign 23) were collected from Women's Hospital, School of Medicine, Zhejiang University from 1995 to 2001. The assessment of MSI was based on the use of a panel of six microsatellite markers (BAT-25, BAT-26, BAT-40, D5S346, D17S250, and D2S123) by polymerase chain reaction (PCR).

[Mismatch repair gene promoter methylation and expression in hydatidiform moles and the malignant transformation].

Objective: In this study, we assayed promoter hypermethylation and protein expression of the mismatch repair gene (MMR) hMLH1 and hMSH2 in gestational trophoblastic diseases to understand the significance of MMR promoter methylation and expression in the pathogenesis and malignant transformation of hydatidiform mole.

Methods: DNA was extracted from chorion of early pregnancies, partial hydatidiform moles, complete hydatidiform moles, and invasive moles were over digested by methylation sensitive endonuclease Hpa II. Then the promoters were amplificated by polymerase chain reaction.