[Expression and promotor methylation of p73 gene in ovarian epithelial tumors].

OBJECTIVE: To investigate the expression and promoter methylation status of p73 gene in ovarian epithelial tumors and their clinicopathological correlations. METHODS: Tissue microarrays (TMA) consisting of 68 ovarian cancers, 37 ovarian borderline tumors and 21 ovarian benign tumors were constructed. p73 expression was detected by immunohistochemistry (EnVision method). Fresh-frozen tissue samples from 13 cases of ovarian carcinomas and 5 cases of borderline tumors were evaluated for the presence of p73 promoter methylation using bisulfite sequencing. RESULTS: Overall, 92.6% (63/68) ovarian carcinomas expressed p73, with a mean value of 32% (percentage of p73 positive cells in the tumor). The mean value of p73 expression rate (40%) in serous carcinoma (26/26) was higher than those of other cancer types (P = 0.006). The mean value of p73 expression rate (40%) in type II ovarian carcinoma was significantly higher than that in type I ovarian carcinoma (24%, P = 0.010). The expression of p73 was not associated with FIGO stage and histological grade (both P > 0.05). The mean values of p73 expression in ovarian borderline tumor (30/37) and benign tumor (12/21) were 16% and 15%, respectively. Of the two groups, the mean value of p73 expression rate in serous type was higher than that in mucous type (P = 0.003, P = 0.026). Ovarian carcinomas had a higher level of p73 expression than borderline tumors and benign tumors (both P < 0.05), while that between ovarian borderline tumors and benign tumors had no statistical difference (P > 0.05). Among serous tumors (49/53), the mean value of p73 expression in the carcinoma group (26/26) was significantly higher than those in the borderline tumor group (12/14) and benign tumor group (11/13; P = 0.024 and P = 0.002, respectively), while that between borderline tumor group and benign tumor group had no statistical difference (P = 0.428). Among mucous tumors (15/27), the mean value of p73 expression in carcinoma group (6/7) was higher than that in benign tumor group (1/8; P = 0.032). No statistical difference of p73 expression was seen between the carcinoma group and ovarian borderline tumor group (8/12) and between the borderline tumor group and benign tumor group (P = 0.234, P = 0.201, respectively). p73 promotor methylation was found in 8 of 13 cases of carcinomas but at different methylation levels with a mean value of 8.0%. Two of 5 ovarian borderline tumors showed detectable p73 promotor methylation with a mean value of 9.0%. Compared with the borderline tumors, ovarian carcinomas showed a similar p73 methylation level (P > 0.05). The p73 methylation level in ovarian carcinomas was not associated with histological type, pathogenetic type, histological grade and FIGO stage (all P > 0.05). CONCLUSIONS: Most of ovarian epithelial tumors express p73 protein with mean values higher in ovarian carcinomas than those in the borderline and benign tumors. Ovarian serous carcinomas have the highest expression level of p73. A simple linear correlation does not exist between the promoter methylation and protein expression of p73.

Overexpression and immunosuppressive functions of transforming growth factor 1, vascular endothelial growth factor and interleukin-10 in epithelial ovarian cancer.

OBJECTIVE: Transforming growth factor-1 (TGF-ß1), vascular endothelial growth factor (VEGF), and interleukin-10 (IL-10) may be critical cytokines in the microenvironment of a tumor, playing roles in immune suppression. This study was conducted to elucidate the roles and immunosuppressive functions of these cytokines in epithelial ovarian cancer (EOC). METHODS: The expression levels of TGF-ß1, VEGF and IL-10 in malignant tissue were evaluated by immune- histochemistry and compared with corresponding borderline, benign, and tumor-free tissues. Moreover, relationships among the levels of these cytokines and correlations between expression and the prognosis of EOC were analyzed by Pearson rank correlations and multi-factor Logistic regression. The roles of TGF-ß1, VEGF, and IL-10 in the immunosuppressive microenvironment of ovarian cancer were studied through dendritic cell (DC) maturation and CD4+CD25+FoxP3+ Treg generation in vitro experiments. RESULTS: TGF-ß1, VEGF, and IL-10 were expressed in 100%, 74.69%, and 54.96% of EOC patients, respectively. TGF-ß1 was an independent prognostic factor for EOC. IL-10 was significantly co-expressed with VEGF. In vitro, VEGF and TGF-ß1 strongly interfered with DC maturation and consequently led to immature DCs, which secreted high levels of IL-10 that accumulated around the tumor site. TGF-ß1 and IL-10 induced Treg generation without antigen presentation in DCs. CONCLUSIONS: TGF-ß1, VEGF and IL-10 play important roles in EOC and can lead to frequent immune evasion events.

[Expression and significance of microRNAs in the p53 pathway in ovarian cancer cells and serous ovarian cancer tissues].

OBJECTIVE: The aim of this study was to investigate whether miR-449a, miR-449b and miR-192 family microRNAs play the same roles in p53 pathway as miR-34 family in ovarian cancer. METHODS: Wild-type p53 ovarian carcinoma cell line A2780 cells were treated with genotoxic agent adriamycin. The reactivation of p53 was detected by Western blot. The expression of miR-449a/b, miR-34a, miR-34b, miR-34c, miR-192 and miR-194 were detected by real-time quantitative PCR. Mutant p53 ovarian cancer cell line SKOV3.ipl cells were transfected with pre-microRNAs and the cell-cycle changes were detected. The expression level of miR-449a/b, miR-34a, miR-34b, miR-34c, miR-192 and miR-194 in serous ovarian carcinomas of varying grade and stage were compared with real-time PCR. RESULTS: The expressions of miR-449a/b, miR-34b and miR-34c were 19-fold to 21-fold elevated after p53 activation by genotoxic agent. Ectopic expression of miR-449b, as well as miR-34c, resulted in cell-cycle arrest in SKOV3.ipl cells. The expression of miR-449a/b was parallel with that of miR-34b, miR-34c, and were significantly lower in late stage and high-grade serous carcinomas than in the normal fallopian tube, early stage and low-grade serous carcinomas. The expression of miR-192, miR-194 and miR-34a did not show evident features in serous ovarian carcinomas and were much lower than miR-449a/b, miR-34b and miR-34c in normal fallopian tube. CONCLUSIONS: As tumor-suppressor microRNAs, miR-449a/b, miR-34b and miR-34c cooperate and play important roles in p53 pathway. Their inactivation may contribute to the carcinogenesis and progression of serous ovarian carcinomas.

[Measurement of serum human epididymis secretory protein 4 combined with CA125 assay in differential diagnosis of endometriosis cyst and ovarian benign and malignant tumors].

OBJECTIVE: To investigate the value of human epididymis secretory protein 4(HE4) combined with CA125 assay in differential diagnosis of endometriosis cyst and ovarian malignant tumor. METHODS: The level of HE4 and CA125 were measured by enzyme-linked immunosorbent assay (ELISA) in the serum specimens of 46 cases in endometriosis cyst group, 36 cases in malignant ovarian tumor group, 60 cases in benign ovarian diseases and 50 women in healthy women group. Those results were shown with median level. The normal range were 0-150 pmol/L in HE4 and 0-35 kU/L, which either one was more than the threshold value defined as positive index. The sensitivity of assay was evaluated by receiver operating characteristic (ROC) curve, the relation and value of HE4 or CA125 alone and combination assay in diagnosis of endometriosis was analyzed by Mann-Whitney U test and correlation analysis. RESULTS: (1) HE4: the median levels of HE4 were 52.4, 51.0, 50.0 pmol/L in group of endometriosis, normal control and benign ovarian tumor, which didn't show statistical difference. However, HE4 was 507.5 pmol/L in ovarian cancer group, which was significantly higher than those of 3 groups (P<0.05). (2) CA125: there were significant different in median level of CA125 was observed as 743.0 kU/L in ovarian cancer, 84.9 kU/L in endoemtriosis, 15.4 kU/L in benign ovarian disease, and 11.5 kU/L in healthy women (P<0.05). (3) The single assay: when compared with that in endometriosis group, receiver operating characteristic area under the curve (ROC-AUC) were 0.933 in HE4 alone and 0.821 in CA125 alone assay in ovarian cancer group. The specificity was 95% and the sensitivity was 79.6% and 49.0%. (4) The combination assay: when compared with those in endometriosis group, the ROC-AUC was 0.936, the specificity was 95% and the sensitivity was 81.0% in ovarian cancer. CONCLUSIONS: Measurement of HE4 could be used in differential diagnosis of endometriosis cyst. And the combination of HE4 and CA(125) assay could discriminate ovarian endometriosis cysts from ovarian malignant tumors effectively.

[Identification of T cell epitopes from ovarian cancer associated anti-idiotype antibody].

OBJECTIVE: To identify the T cell epitopes from ovarian cancer associated anti-idiotypic antibody 6B11 in order to explore the molecular basis of 6B11 induced cellular immune responses against ovarian cancer. METHODS: Potential human leukocyte antigen (HLA) A0201 ligands were predicted by using SYFPEITHI algorithm and tested by the T2 binding assay for screening of HLA-A2 binding peptides from 6B11 complimentary determining region (CDR). Cytotoxic T lymphocytes (CTL) to 6B11 or peptides were generated by 3 rounds of in vitro stimulation with 6B11 or peptide-pulsed dendritic cells (DC), and then tested by (51)Cr-release assay to ascertain the CTL epitope of 6B11. Cell proliferation assay was performed by using 6B11 specific CTL as responder cells and peptide-pulsed DC as stimulators to ascertain the helper T lymphocyte (Th) epitope of 6B11. Cytokine assay and interferon-gamma ELISPOT assay were performed to verify the CTL and Th epitope of 6B11 further. RESULTS: Light chain CDR3 peptide (VL CDR3) of 6B11 induced specific CTL to kill HLA-A2 and target antigen positive ovarian cancer cells, which could be blocked by anti human major histocompatibility complex (MHC) I antibody. Heavy chain CDR3 peptide (VH CDR3) of 6B11 stimulated the proliferation of 6B11-primed CTL, which could be blocked mainly by anti human MHC-II antibody, and further experiments showed that part of the VH CDR3 peptide-primed CTL killed K562 cells. Peptides in VL CDR3 and VH CDR3 were the CTL and Th epitope mimicking the original antigen of 6B11 respectively. Collaboration of 6B11 CTL and Th epitope, or 6B11 CTL epitope and keyhole limpet hemocyanin (KLH), the former was more powerful in inducing specific cellular immune responses against ovarian cancer. 6B11 or corresponding CTL and Th epitope specific CTL secreted high levels of interleukin-2 (1630, 1503 ng/L) and interferon-gamma (5620, 5421 ng/L), while basal level of interleukin-4 was detected (253, 274 ng/L). ELISPOT assay confirmed the existence of specific interferon-gamma secreting cells in 6B11 or epitopes specific CTL (196/1 x 10(6) T cell, 184/1 x 10(6) T cell). CONCLUSIONS: VL CDR3 and VH CDR3 peptides of 6B11 are the CTL and Th epitopes mimicking original antigen which could duplicate the activity of intact 6B11 to induce the cellular immune responses against ovarian cancer. The results have significant theoretical and practical value in application of anti-idiotypic antibody as anti tumor vaccine. The acquired CTL and Th epitopes as constituents of future multiple peptides against ovarian cancer could be used in peptide vaccine based ovarian cancer therapy.

[Effect of reduced expression of Her2 by RNA interference on the biological characters of ovarian carcinoma cells].

OBJECTIVE: To observe the effects of short hairpin RNA (shRNA) targeting Her2 on its gene expression when the shRNA was stably transfected into human ovarian cell lines, SKOV3 and SKOV3.ip1, which have different extent of malignancy and investigate the changes of the biological characters of the two cell lines after the stable transfection. METHODS: The plasmids expressing shRNA targeting Her2 gene were transfected into SKOV3 and SKOV3.ip1 cells. The stably transfected cells were gained by antibiotic screening. The expression of Her2 before and after the transfection was detected by RT-PCR and western blot. The transwell experiment was used to observe the invasion ability of the cancer cells before and after the transfection, and the parent and the transfected cells were injected into nude mice to observe the tumor growth. RESULTS: After the stable transfection with Her2 shRNA, mRNA and protein levels of Her2 gene in SKOV3 and SKOV3.ip1 cells were remarkably reduced. The expression of mRNA were (68.0 +/- 3.1)%, (40.8 +/- 2.0)%, (99.9 +/- 1.3)%, (42.4 +/- 2.5)%. The expression of protein were (72.1 +/- 3.4)%, (36.4 +/- 1.5)%, (98.2 +/- 1.7)%, (40.7 +/- 2.1)%. The invasion ability into basilar membrane of the transfected cells was greatly reduced compared with the parent cells. The invasion cell numbers were 7.6 +/- 1.1, 1.8 +/- 0.8, 36.2 +/- 9.7, 15.7 +/- 7.2. The growth rate of the planted tumors was lower in transfected groups than that of the parent groups. CONCLUSIONS: (1) The expression of Her2 gene in SKOV3 and SKOV3.ip1 cells was remarkably reduced by RNA interference targeting Her2. (2) The biological characters of SKOV3 and SKOV3.ip1 cells are changed when the expression of Her2 gene is reduced by RNA interference.

[The values of serum human epididymis secretory protein 4 and CA(125) assay in the diagnosis of ovarian malignancy].

OBJECTIVE: To evaluate the value of human epididymis secretory protein 4 (HE4) and CA(125) in the diagnosis of ovarian malignancy. METHODS: HE4 and CA(125) in the serum specimens of malignant ovarian tumor group (30 cases), benign ovarian diseases (110 cases; 45 benign ovarian tumor, 57 endometriotic diseases and 8 pelvic inflammation were included) and healthy women group (137 cases) were assayed double blindly. The levels and the diagnosis efficiency of the HE4 and CA(125) were analyzed. RESULTS: (1) The median levels of HE4 and CA(125) were significantly higher in malignant ovarian tumor group (244 pmol/L and 601 kU/L respectively) than those of the benign ovarian diseases group (32 pmol/L and 22 kU/L respectively) and healthy women group (32 pmol/L and 11 kU/L respectively) (P = 0.000 - 0.029). The median levels of CA(125) were also higher in endometriotic diseases and pelvic inflammation groups (53 and 41 kU/L respectively) than those of benign ovarian tumor group and healthy women group (12 and 11 kU/L respectively; P = 0.000 - 0.031). (2) The positive rate of HE4 was lower than that of CA(125) in malignant ovarian tumor group (P = 0.036). HE4 was negative in benign diseases and healthy women groups. But the positive rates of CA(125) were 56.1% and 5/8 respectively in endometriotic diseases and pelvic inflammation groups and there were significant differences compared with HE4 (P = 0.000). (3) The HE4 assay had advantage over the CA(125) assay in receiver operating characteristic-area under the curve (ROC-AUC) and sensitivity with a specificity of 100% when ovarian malignancy was compared with controls having benign diseases and healthy women, benign tumor or benign diseases groups respectively. The CA(125) assay had advantage over the HE4 assay in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having healthy women group. (4) Combined assay of HE4 and CA(125) was better than CA(125) alone when ovarian malignancy was compared with controls having any group. (5) Combined assay was better than HE4 alone in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having benign diseases and healthy women or healthy women groups. And combined assay was lower in the ROC-AUC and the sensitivity with specificity of 100% than HE4 when ovarian cancers were compared with controls having benign tumors or benign diseases groups respectively. (6) The diagnosis efficiency of the HE4 assay at the level 86 pmol/L determined in ROC curve with controls having benign diseases and healthy women group and at the 95% reference level 50 pmol/L of healthy women or 150 pmol/L recommended by the kit respectively was compared. The sensitivity of 50 pmol/L was 73% higher than 150 pmol/L and 86 pmol/L, while the specificity and positive predictive value were lower (P = 0.002, P = 0.000). The specificity, accuracy and positive predictive value of HE4 assay at the set point of 150 pmol/L and 86 pmol/L were 100%, 96% and 96%. The set point of 86 pmol/L had advantage over 150 pmol/L at the sensitivity of diagnosis, 70% and 63% respectively. But the positive predictive value was 95% lower than 150 pmol/L, being 100%. There was no significant difference (P = 0.883, P = 0.883). CONCLUSIONS: The specificity of HE4 assay is higher than CA(125) assay in the diagnosis of ovarian cancer and HE4 combined with CA(125) assay can improve the diagnoses. The set point of 150 pmol/L is advantageous for the accurate diagnosis, while the set point of 86 pmol/L is advantageous for the screening of malignant ovarian cancer.

[Construction and utilization of the prognostic model of serous ovarian adenocarcinoma].

OBJECTIVE: To analyze the related factors with prognosis in patients with serous ovarian adenocarcinoma and to set up a prognostic model of serous ovarian adenocarcinoma. METHODS: The clinical, pathological and follow-up data of 104 cases with serous ovarian adenocarcinoma were retrospectively analyzed. Kaplan-meier univariate analysis was used to screen the prognostic factors; COX univariate and multivariate analyses were used to determine the risk coefficient of each factors and different layers in each factor. Pearson rank correlation was used to reject the influence of different factors with each other. And the prognostic model of serous ovarian adenocarcinoma was set up based on the result of the above study, which could be used to deduce the survival probability of patients with serous ovarian adenocarcinoma. RESULTS: International Federation of Gynecology and Obstetrics (FIGO) stage (P = 0.0029), histological grade (P = 0.0054), residual disease (P = 0.0000), metastasis of lymph nodes (P = 0.0000) and chemotherapy (P = 0.0000) were the related factors of prognosis in patients with serous ovarian adenocarcinoma, of which FIGO stage was the most important one, followed sequentially by histological grade, metastasis of lymph node, residual disease and chemotherapy (the independent risk coefficient of each factor was 1.3392, 0.9206, 0.7071, 0.6004, 0.4985 in sequence). We set up a prognosis model according to the prognostic index of each factors. The effect of chemotherapy and residual disease on prognosis could be quantified by this model, and the higher the score, the lower the survival probability of patients. CONCLUSIONS: FIGO stage, histological grade, residual disease, metastasis of lymph nodes and chemotherapy are important prognostic factors of serous ovarian adenocarcinoma. This model can be used to estimate the prognosis of patients with serous ovarian adenocarcinoma, and the effect of both chemotherapy and residual disease on the prognosis could be quantified by the model.

[In vitro study of the antitumor immune responses induced by anti-idiotypic minibody vaccine of ovarian cancer].

OBJECTIVE: To evaluate whether anti-tumor immune response can be induced in vitro with 6B11 anti-idiotypic minibody. and to explore its probability as ovarian cancer vaccine. METHODS: Separated human peripheral blood mononuclear cells (PBMC) were stimulated and cultured by 6B11 minibody. The proliferations of PBMC and cytotoxin were observed by (3)HTdR and (51)Cr release test respectively. ELISA(Enzyme-Linked Immunosorbent Assay) test and Immune Flow Cytometry were used to analyze IFN-gamma in supernatant of the cultured cdlls and the change of T lymphocyte phenotype of PBMC with 6B11 minibody stimulated. RESULTS: 6B11 minibody could stimulate PBMC to proliferate, the best dose was 20 mg/L; it performed cytotoxin function to ovarian carcinoma cell line expressing OC166-9. IFN-gamma maintained at high level after stimulation. It stimulated proliferation of CD3(+) T cell and CD4(+) from PBMC after stimulation respectively. CD8(+) T cell proliferation was not clear. There was significant difference between stimulation and unstimulation in CD4(+)/CD8(+) ratio. CONCLUSION: 6B11 anti-idiotypic minibody can induce both humoral and cellular immunity against ovarian carcinoma in vitro. This paper has provided strong experimental evidence for clinical use of 6B11 minibody as anti-idiotype vaccines against ovarian carcinoma.

[Monitoring novel ovarian carcinoma associated genes using cDNA expression microarray].

OBJECTIVE: To explore the gene expression pattern of sample of human ovarian carcinoma. METHOD: The difference in gene expression between normal and neoplastic human ovarian tissues were investigated, we described the assembly and utilization of a 512 member cDNA microarray. RESULT: Thirty-seven genes expressed in ovarian cancer were screened out, 14 genes were up-regulated, 23 genes were down-regulated. CONCLUSION: cDNA microarray for analysis of gene expression pattern is an effective method to identify novel ovarian cancer associated genes.

Effect of human epididymis protein 4 gene silencing on the malignant phenotype in ovarian cancer.

BACKGROUND: Human epididymis secretory protein 4 (HE4) has been proved to be a promising novel biomarker for the detection of epithelial ovarian carcinomas. Compared with CA125, HE4 assay demonstrated an improved ability to discriminate between pelvic mass with malignant and benign disease. Though it is well known that HE4 is overexpressed in ovarian cancer, however, the role of HE4 in the carcinogenesis and progression of ovarian cancer remains unkown. METHODS: In this study, we explored the role of HE4 in the carcinogenesis and progression of ovarian cancer. We screened nine ovarian cancer cell lines for HE4 expression, and using RNA interference (RNAi), we silenced HE4 gene expression in CaoV3 and SKOV3.ip1 ovarian cancer cell lines. We assessed the effect of HE4 gene silencing on the transformed phenotype by examining the cell cycle, apoptosis, proliferation and transwell migration/invasion in vitro. RESULTS: HE4 gene silencing induces G0/G1 arrest and blocks the progression from the G1 to S phase in CaoV3 and SKOV3.ip1 cells. HE4 knockdown also inhibited cell proliferation, migration and invasion in SKOV3.ip1 cells in vitro. CONCLUSION: HE4 may be involved in the regulation of the cell cycle and promote ovarian cancer migration and invasion.

Screening for genes associated with ovarian cancer prognosis.

BACKGROUND: Human epithelial ovarian cancer cell line SKOV3.ip1 is more invasive and metastatic compared with its parental line SKOV3. A total of 17 000 human genome complementary DNA microarrays were used to compare the gene expression patterns of the two cell lines. Based on this, the gene expression profiles of 22 patients with ovarian cancer were analyzed by cDNA microarray, and screened the 2-fold differentially expressed genes compared with the normal ones. We screened genes relevant to clinical prognosis of serous ovarian cancer by determining the expression profiles of ovarian cancer genes to investigate cell receptor and immunity-associated genes, and as groundwork, identify ovarian cancer-associated antigens at the gene level. METHODS: Total RNA was extracted from 22 patients with ovarian cancer and DNA microarrays were prepared. After scanning, hybridization signals were collected and the genes that were differentially expressed twice as compared with the normal ones were screened. RESULTS: We screened 236 genes relevant to the prognosis of ovarian cancer from the 17 000 human genome cDNA microarrays. According to gene classification, 48 of the 236 genes were cell receptor or immunity-associated genes, including 2 genes related to the International Federation of Gynecology and Obstetrics (FIGO) stage, 4 genes to histological grade, 18 genes to lymph node metastasis, 11 genes to residual disease, and 13 genes to the reactivity to chemotherapy. Several functionally important genes including fibronectin 1, pericentriolar material 1, beta-2-microglobulin, PPAR binding protein were identified through review of the literature. CONCLUSIONS: The cDNA microarray of ovarian cancer genes developed in this study was effective and high throughput in screening the ovarian cancer-associated genes differentially expressed. Through the studies of the cell receptor and immunity-associated genes we expect to identify the molecular biology index of ovarian cancer-associated antigens.

[Specific immune cell therapy against ovarian cancer in vivo and in vitro].

BACKGROUND & OBJECTIVE: 6B11minibody (6B11mini), an anti-idiotypic vaccine against human ovarian cancer, has been proven to induce specific humoral and cellular immunity against ovarian cancer in vivo and in vitro. This study was to investigate the safety and efficacy of using 6B11mini as an antigen to treat ovarian cancer. METHODS: After being loaded with purified 6B11mini, dendritic cells (DCs) were co-cultured with peripheral blood mononucleocytes (PBMNC) and stimulated by various cytokines, including CD3 monoclonal antibody,interleukin-2, interferon-gamma, to obtain 6B11mini-ovarian-cytokine-induced-killer cells (6B11-O-CIK). Tumor-forming ability was determined using soft agar colony-forming assay in vitro and nude mice xenografts in vivo. The acute toxicity of 6B11-OCIK at different doses was observed in BALB/c mice. Cytotoxicity of 6B11-OCIK to different target cells was detected using 51Cr release test in vitro. The ovarian tumor model was established using severe combined immune deficiency (SCID) mice transplanted with human ovarian cancer cell line SKOV3. The tumor growth was detected after injection of 6B11-OCIK into SCID mice. Injection of CIK, PBMNC and physiological saline were used as controls. RESULTS: After a cultural period of 14 days in soft agar, SKOV3 cell clones were well formed with a ratio of 50%; while 6B11-OCIK, CIK and PBMNC did not form any clones. All nude mice injected with human cervical carcinoma cell line HeLa (positive control) grew tumors after 14 days, and mice injected with 6B11-OCIK, CIK, PBMNC and normal human fetal lung fibroblast WI-38 cells did not form tumors after 13 weeks. BALB/c mice did not show any abnormal response half an hour after the administration of 6B11-OCIK cells at different doses. Mice were sacrificed 13 days after treatments, but no distinct abnormality of the main organs were found. 6B11-OCIK exerted specific cytotoxicity against tumor cells with positive OC166-9, which was related to the limitation of MHC. The tumor weights of SCID mice transplanted with SKOV3 cells were significantly lighter in 6B11-OCIK treatment group than in the saline group(P=0.023); while the tumor weights were not significantly different between the 6B11-OCIK group with CIK and the PBMNC group(P=0.540; P=0.285). CONCLUSIONS: The application of 6B11-OCIK in vivo has reached the safety standard. 6B11-OCIK has the inhibitory effect on the growth of ovarian cancer cells.

[Human pituitary tumor transforming gene 1 expression in ovarian carcinoma and its clinical significance].

BACKGROUND & OBJECTIVE: Human pituitary tumor transforming gene 1 (hPTTG1) is a newly identified oncogene. We performed a large-scale screening survey to identify related genes expressed in advanced and poorly differentiated serous ovarian carcinoma using cDNA microarray analysis, and found that hPTTG1 is one of highly up-regulated genes in ovarian carcinoma. This study was to examine hPTTG1 mRNA and protein expression in various epithelial ovarian carcinoma,analyze the relationship of its expression level with FIGO stage and histological grade, and explore the functional role of hPTTG1 in ovarian carcinoma pathogenesis. METHODS: HPTTG1 mRNA expression in 27 cases of epithelial human ovarian carcinoma, and 4 cases of normal ovary was assessed with non-competitive reverse transcription polymerase chain reaction (RT-PCR), and hPTTG1 protein expression in these 27 cases of human ovarian carcinoma, and 18 cases of normal ovary was examined by immunohistochemistry. RESULTS: HPTTG1 mRNA expression level in normal ovary was low, while in ovarian carcinoma was significantly higher than that in normal counterparts (P < 0.01). Fold increase of ovarian cancer tissues to normal ovary tissues is 1.1-4.8 (median 2.4).HPTTG1 mRNA high expression level was related to poor tumor differentiation (r = 0.686, P < 0.05), but the correlation with FIGO stage was not detected. HPTTG1 protein was expressed in all cases of ovarian carcinoma, but not in normal ovaries. CONCLUSIONS: Increased hPTTG1 expression may be an important factor involved in early tumorogenesis, and may be associated with poor tumor differentiation. HPTTG1 could be a potential molecular marker of epithelial ovarian carcinoma.