The cellular inhibitor of apoptosis protein-2 is a possible target of novel treatment for endometriosis.

PROBLEM: How is the tumor necrosis factor (TNF) α-induced inhibitor of apoptosis (IAP) protein expression in endometriotic stromal cells (ESCs) involved in cell viability and signaling pathways? METHOD OF STUDY: Endometriotic stromal cells were isolated from ovarian chocolate cysts in 20 patients who underwent laparoscopic surgery. IAP protein expression and IκB phosphorylation were evaluated by Western blot analysis. Interleukin (IL)-8 protein expression and cell proliferation were assessed by ELISA. RESULTS: Cellular IAP (cIAP)-2 protein expression in endometriotic tissue was higher than that of endometrium. TNFα markedly enhanced cIAP-2 protein expression in ESCs. Pretreatment with a nuclear factor (NF)-κB inhibitor attenuated TNFα-induced cIAP-2 expression. An antagonist of IAPs abrogated TNFα-induced cIAP-2 protein expression and showed a decrease in TNFα-induced IL-8 protein expression and BrdU incorporation in ESCs. CONCLUSIONS: TNFα and its downstream NFκB pathway have proven to be critical regulators of highly expressed cIAP-2 in ESCs. cIAP-2 may be a novel therapeutic target for endometriosis.

Ultrasound for evaluation of adnexal malignancy: from 2D to 3D ultrasound.

Conventional two-dimensional (2D) ultrasound has been widely used for the evaluation of adnexal malignancy in gynecologic fields. This 2D ultrasound evaluation includes a morphological assessment, color/power and pulsed Doppler sonographic assessment, scoring system, and contrast agent assessment of adnexal masses. The introduction of three-dimensional (3D) ultrasound would facilitate the novel assessment of adnexal masses. With the recent advance in 3D power Doppler (3DPD) ultrasound as well as quantitative 3DPD histogram analysis, quantitative and qualitative assessments of the vascularization and blood flow of adnexal masses have become feasible. These novel techniques may assist in the evaluation of adnexal malignancy, and offer potential advantages relative to conventional 2D sonographic assessments. 3D ultrasound may be an important modality in future gynecologic oncology research and in the evaluation of adnexal malignancy in clinical practice, although some limitations regarding the assessment of adnexal malignancy employing 3D ultrasound remain unresolved.

Expression of the vascular endothelial growth factor (VEGF) gene in epithelial ovarian cancer: an approach to anti-VEGF therapy.

AIM: A monoclonal antibody that targeted vascular endothelial growth factor (VEGF) resulted in a dramatic suppression of tumor growth in vivo, which led to the development of bevacizumab, a humanized variant of anti-VEGF antibody, as an anticancer agent. The aims of this study were to clarify the significance of VEGF gene expression in relation to clinicopathological parameters and to identify potential candidates for anti-VEGF therapy with bevacizumab. PATIENTS AND METHODS: VEGF gene expression was analyzed by real-time quantitative reverse transcription-polymerase chain reaction in 178 surgical epithelial ovarian cancer specimens. This gene expression was correlated with clinicopathological parameters and patient survival. RESULTS: The median VEGF gene expression level and range relative to GAPDH were 0.147 and 0.016-2.44, respectively. Patients were dichotomized into two groups with low and high expression levels by using the median value as the cutoff. VEGF gene expression did not affect prognosis of patients overall (p = 0.541). Although statistical significance was not noted, we found the prognosis of patients with high VEGF gene expression tended to be worse than that of those with low VEGF gene expression by univariate Cox regression analysis (p = 0.085) in patients with stage III-IV cancer. Macroscopic residual disease (positive; p = 0.012) was significantly associated with poor prognosis in univariate Cox regression analysis in patients with stage III-IV cancer. Moreover, presence of macroscopic residual disease was positively associated with VEGF gene expression (p = 0.030) in patients with stage III-IV cancer. CONCLUSION: Patients with epithelial ovarian cancer with tumors with positive macroscopic residual disease and high VEGF gene expression could be potential candidates for anti-VEGF therapy with bevacizumab.

Association of metastin/a G-protein-coupled receptor signaling and Down syndrome critical region 1 in epithelial ovarian cancer.

UNLABELLED: It has been revealed that metastin/a G-protein-coupled receptor (AXOR12) signaling enhances the expression of Down syndrome critical region 1 (DSCR1), known to be duplicated in Down syndrome, and suppresses tumor metastasis in in vitro study. The aim of this study was to evaluate whether gene expression of metastin/AXOR12 signaling system is correlated with that of DSCR1 and consequently affect prognosis of patients with epithelial ovarian cancer. PATIENTS AND METHODS: The expression levels of metastin, AXOR12, DSCR1 isoform 1 (DSCR1-1), DSCR1 isoform 4 (DSCR1-4), calcineurin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression were analyzed by real-time quantitative reverse transcription-polymerase chain reaction in 102 epithelial ovarian cancer surgical specimens. RESULTS: Patients were dichotomized into two groups having low and high expressions by using the median value as the cut-off A good agreement was noticed between metastin and AXOR12 gene expression levels (kappa coefficient; 0.73), however, the gene expression of metastin/AXOR12 signaling system was not significantly correlated with that of DSCR1-4. By univariate Cox regression analysis, the prognosis of the patients with low metastin and low AXOR12 gene expression was significantly worse than that of those with high metastin and high AXOR12 gene expression, respectively (p = 0.04 and 0.018). Combination of metastin and AXOR12 gene expression also had significant impact on patient prognosis (p = 0.045). The DSCR1-1, DSCR1-4 and calcineurin gene expressions did not significantly affect the prognosis. CONCLUSION: The precise mechanism of metastin/ AXOR12 signaling for suppression of the invasive phenotype in vivo, especially in epithelial ovarian cancer, is still uncertain. Genes such as DSCR1 that are duplicated in Down syndrome might not play an important role in tumorigenesis of epithelial ovarian cancer.

Positron emission tomography with the glucose analog [F]-fluoro-2-deoxy-D-glucose for evaluating pelvic lymph node metastasis in uterine corpus cancer: comparison with CT and MRI findings.

OBJECTIVE: The aims of this study were to investigate the ability of positron emission tomography (PET) with the glucose analog [(18)F]-fluoro-2-deoxy-D-glucose (FDG) to detect pelvic lymph node metastasis of uterine corpus cancer and to perform a retrospective comparison with computed tomography (CT) and magnetic resonance imaging (MRI) findings. METHODS: Forty-six patients with uterine corpus cancer scheduled for surgery, including bilateral pelvic and/or para-aortic lymphadenectomy, were eligible for this study. CT and MRI of the pelvis and abdomen were performed in all patients within 2 weeks preceding whole-body FDG-PET. FDG-PET images were analyzed visually for objective assessment of regional tracer uptake. The sensitivity and specificity of each imaging modality for detecting pelvic lymph node metastasis were determined, respectively. RESULTS: Eleven (7 with myometrial invasion less than 1/2, 4 with myometrial invasion over 1/2) of the 46 patients (23.9%) were revealed to have pelvic lymph node metastasis. The sensitivity and specificity for detecting pelvic lymph node metastasis in patients with uterine corpus cancer by FDG-PET were 31.3% and 96.1% by lymph node region, respectively, and 36.4% and 91.4% by patient, respectively. No significant difference was noted among each imaging modality with sensitivity or specificity. Moreover, the sensitivity and specificity for detecting pelvic lymph node metastasis in 29 patients with endometrioid adenocarcinomas by FDG-PET were 28.6% and 96.1% by lymph node region, respectively, and 50% and 92% by patient, respectively. No significant difference was noted among each imaging modality in terms of sensitivity or specificity. Among 11 patients with pathologically positive pelvic lymph node metastasis, three out of four patients with myometrial invasion over 1/2 were diagnosed as pelvic lymph node metastasis positive by all diagnostic modalities, however, only one of seven patients with myometrial invasion less than 1/2 was diagnosed as pelvic lymph node metastasis positive by PDF-PET and CT. MRI could not detect pelvic lymph node metastasis in patients with myometrial invasion less than 1/2. CONCLUSIONS: We suggest that current imaging modalities including FDG-PET cannot change medical management of patients with uterine corpus cancer before surgery.

Does three-dimensional power Doppler ultrasound improve the diagnostic accuracy for the prediction of adnexal malignancy?

OBJECTIVE: The aim of the present study was to investigate the diagnostic accuracy of three-dimensional power Doppler ultrasound (3DPD) in the differentiation between benign and malignant adnexal masses and evaluate 3DPD for assessing malignancy in comparison with two-dimensional transvaginal gray-scale sonography (2DTVS), magnetic resonance imaging (MRI) and positron emission tomography (PET). SUBJECTS AND METHODS: Thirty-six patients with suspicious adnexal masses were included in this study. 2DTVS and 3DPD were performed preoperatively. MRI and PET examinations were also carried on within two weeks of initial sonography. All the results of diagnostic imaging techniques were evaluated separately. Final diagnosis was confirmed by the postoperative histopathology. The mature cystic teratomas and endometriomas with typical gray-scale sonographic appearance were excluded from the study. RESULTS: Of the 36 patients, 25 had a malignancy, 5 had a borderline tumor, and 6 had a benign mass. Sensitivity of 2DTVS, 3DPD, MRI and PET were 96.7%, 76.7%, 96.7% and 83.3%, respectively. The sensitivity of 2DTVS was as high as that of MRI, and both of them were significantly higher than those of 3DPD and PET, respectively. There were no significant differences both in specificity and accuracy among these 4 techniques. CONCLUSION: 3DPD did not improve the diagnostic accuracy for the prediction of malignancy in adnexal masses. 2DTVS may still remain an important modality for the prediction of adnexal malignancy.

Expression of metastin and a G-protein-coupled receptor (AXOR12) in epithelial ovarian cancer.

BACKGROUND: Metastin, a product of the KiSS-1 gene, is a ligand for a G-protein-coupled receptor (AXOR12) and is a strong suppressant of metastasis. The aim of this study was to evaluate whether metastin and AXOR12 gene expressions affect prognosis of patients with epithelial ovarian cancer. METHODS: The expression levels of metastin, AXOR12 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression were analysed by the real-time quantitative reverse transcription-polymerase chain reaction in 76 epithelial ovarian cancer surgical specimens. Their expression (metastin/GAPDH and AXOR12/GAPDH ratios) was correlated with the clinical findings. Furthermore, cellular distribution of metastin and AXOR12 mRNA was examined by in situ hybridisation on tissue sections. RESULTS: The median and range of mRNA expression for metastin and AXOR12 were 0.047 and 0.01-13.57, and 4.00 and 0.011-135.13, respectively. Patients were dichotomised into two groups having low and high expressions by using the median value as the cutoff. A good agreement was noticed between metastin and AXOR12 gene expression levels (kappa coefficient; 0.74). The presence of residual tumour following resection was negatively associated with metastin (P=0.0084) and AXOR12 (P=0.0148) gene expressions indicating an association of low expression of these genes in more aggressive, and advanced tumours. By univariate Cox regression analysis, the prognosis of the patients with low AXOR12 gene expression was significantly worse than those with high AXOR12 gene expression (P=0.030). The combination of metastin and AXOR12 gene expression level was also significantly associated with the prognosis (P=0.049). Transcripts for both metastin and AXOR12 were detected in the epithelial ovarian carcinoma cells. CONCLUSIONS: These results present a new insight into the understanding of the biological behaviour of epithelial ovarian cancer. Metastin/AXOR12 signalling may suppress the invasive phenotype of epithelial ovarian cancer.

Differential expression of RUNX genes in human esophageal squamous cell carcinoma: downregulation of RUNX3 worsens patient prognosis.

BACKGROUND: The RUNX proteins are a family of transcriptional factors that have essential functions during embryogenesis and development, whereas deregulation in expression of RUNXs is often linked to tumor formation. To date, there has been no study describing the precise expression, prognostic impact and methylation status of RUNXs in esophageal squamous cell carcinoma. METHODS: Resected specimens from 61 patients with esophageal SCC were used to identify the expression of RUNX1, RUNX2 and RUNX3 by real-time RT-PCR. Localization of mRNA was done by in situ hybridization, expression of Smad4 was evaluated by immunohistochemistry, and the methylation status of RUNX3 was analyzed by methylation-specific PCR. RESULTS: RUNX3 had a significant impact on patient prognosis with worse survival in the RUNX3-negative group. In early tumors (T1/T2), the prevalence of lymph vessel invasion and the number of metastatic lymph nodes were significantly higher in RUNX3-negative tumors. Furthermore, RUNX3 became a strong prognostic factor only in Smad4-positive tumors. Also, the methylation status of the RUNX3 promoter had a significant correlation with the loss of RUNX3 expression. CONCLUSION: Downregulation of RUNX3 may play a role in disease progression of esophageal SCC, and hypermethylation of the promoter region might be one of the crucial pathways to silence RUNX3 gene.

Adenocarcinoma in situ coexisting with carcinoma in situ of the cervix during pregnancy.

CASE REPORT: We report a pregnant patient with adenocarcinoma in situ (AIS) coexisting with carcinoma in situ (CIS) of the cervix diagnosed by conization at 16 weeks' gestation. Apoptotic activity was higher in the CIS lesion than in the AIS lesion in the cone biopsy specimen. Postpartum evaluation confirmed the disappearance of CIS lesion with positive cone margins, however, multifocal AIS with negative cone margins was found. CONCLUSION: Clinical course and biological features of AIS associated with pregnancy may be different from those of CIS.

Expression of the angopoietin-1, angopoietin-2, Tie2, and vascular endothelial growth factor gene in epithelial ovarian cancer.

OBJECTIVES: Angiopoietin/Tie2 system with vascular endothelial growth factor (VEFG) is known to be important for the initiation of angiogenesis in tumors. The aim was to evaluate whether angiopoietin/Tie2 system with VEFG affects prognosis in patients of epithelial ovarian cancer. METHODS: Angiopoietin-1 (Ang-1), Angiopoietin-2 (Ang-2), Tie2, and VEGF gene expression were analyzed by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) in 85 epithelial ovarian cancer surgical specimens. These gene expressions were correlated with clinical-pathological parameters, microvessel density (MVD), and patients' survival. RESULTS: Ang-1/Ang-2 gene expression ratio, VEGF, and Tie2 gene expression significantly associated with MVD, respectively (P < 0.0001, P = 0.024, P = 0.005). The patients with low Ang-1/Ang-2 gene expression ratio and high VEGF gene expression were found to have a significantly higher MVD when compared to others (P = 0.0003). Moreover, there was a significant difference between the values of MVD in patients with low Ang-1/Ang-2 gene expression ratio and high VEGF and high Tie2 gene expression and those in others (P = 0.0025). FIGO stage (P = 0.014), residual disease (P = 0.042), histological grade (P = 0.028), Ang-1/Ang-2 gene expression ratio (P = 0.010), and combination of Ang-1/Ang-2 gene expression ratio and VEGF gene expression (P = 0.019), were found to be significantly associated with a poor prognosis in univariate Cox regression analysis. Multivariate Cox regression analysis revealed that FIGO stage is an independent prognostic factor (P = 0.035). Low Ang-1/Ang-2 gene expression ratio had a tendency to be an independent prognostic factor (P = 0.061). CONCLUSIONS: Angiogenesis occurred by angiopoietin/Tie2 system in concert with VEGF in epithelial ovarian cancer did not affect patients' survival. However, gene expression of Ang-1 and Ang-2 might present a pertinent diagnostic tool to select a high-risk group of patients independent of clinical-pathological parameters and a new insight to understand the biology of epithelial ovarian cancer.

Evaluation of the antiangiogenic effect of Taxol in a human epithelial ovarian carcinoma cell line.

PURPOSE: Angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) are major ligands for the endothelium-specific tyrosine kinase receptor Tie-2 and are important regulators of endothelial cell survival. In the presence of vascular endothelial growth factor (VEGF), vessel destabilization by Ang-2 has been hypothesized to induce an angiogenic response, but in the absence of VEGF, Ang-2 leads to vessel regression. In the present study, a human ovarian cancer cell line was used to investigate the possibility that Taxol might affect the expression of Ang-1, Ang-2, and VEGF. MATERIALS AND METHODS: KF 28, a single-cell clone of a human ovarian epithelial carcinoma cell line, was used. The expression of Ang-1, Ang-2, and VEGF was assessed by quantitative real-time RT-PCR and Western blot analysis or enzyme-linked immunosorbent assay. Conditioned medium was used in the in vitro angiogenesis assay. RESULTS: The concentration of Taxol that inhibited the growth of cells to the level of 50% of control cell growth was 4.65+/-0.35 nM. Quantitative real-time RT-PCR indicated that Ang-1 gene expression was significantly decreased by exposure to 2 nM Taxol for 168 h ( P<0.05 vs control cells). Western blot analysis confirmed that the Ang-1 protein level was decreased by exposure to 2 nM Taxol for 168 h. Ang-2 gene expression did not significantly differ between control cells and those exposed to Taxol for any of the indicated times. The Ang-1/ Ang-2 gene expression ratio was significantly decreased by exposure to Taxol for 168 h ( P<0.05 vs control cells). VEGF gene expression was significantly decreased by exposure to Taxol for 168 h ( P<0.05). The VEGF concentration in the conditioned medium was also significantly reduced by exposure to Taxol for 168 h ( P<0.05). Conditioned medium collected following Taxol treatment for 168 h significantly inhibited endothelial tubule formation ( P<0.05). Cell growth did not significantly differ between control cells and those exposed to Taxol for any of the indicated times. CONCLUSIONS: Our results show that exposure of ovarian cancer cells to a low concentration of Taxol may inhibit the initiating event in angiogenesis, namely, vascular regression. This information might be valuable in the development of new therapeutic interventions for epithelial ovarian cancer.

Color Doppler imaging and angiogenesis in ovarian tumors.

The detection and assessment of ovarian lesions are an important part of gynecological practice. Advance knowledge of whether an overt ovarian mass is malignant may be useful for improving the effectiveness of surgical treatment. Pelvic ultrasonography has been developed continuously, and (in conjunction with biochemical and genetic markers) is being used to achieve these objectives with increasing accuracy. At the same time, knowledge is being accrued about the relationship between ultrasound-derived indices of ovarian tumor vascularity and the molecular aspects of angiogenesis. There is now the exciting prospect that color Doppler imaging with pulsed Doppler spectral analysis could also be used to monitor the development and intermediate effectiveness of novel anticancer treatment.

Dihydropyrimidine dehydrogenase in normal and malignant endometrium: relationship with cell proliferation and thymidine phosphorylase.

Dihydropyrimidine dehydrogenase (DPD) is a pyrimidine salvage enzyme responsible for degradation of thymine, which is produced from thymidine by thymidine phosphorylase (TP). Our purpose was to determine the relationship between DPD, cell proliferation and TP expression in human endometrium. We examined DPD gene expression using reverse transcription-polymerase chain reaction, DPD protein levels using enzyme-linked immunosorbent assay, and DPD protein localization using immunohistochemistry in 58 normal endometria and 28 endometrial cancers. DPD gene expression was then related to the proliferating cell nuclear antigen index and to TP gene expression. DPD gene expression, which was correlated with DPD protein level, was relatively stable throughout various menstrual phases but was significantly elevated in postmenopausal status. It was significantly lower in endometrial cancer than in normal endometrium. Localization analysis revealed that DPD protein was located primarily in epithelial cells, but was also present in stromal cells. DPD gene expression correlated inversely with the PCNA index. TP gene expression pattern contrasted with that of DPD in postmenopausal and malignant endometrium. A high ratio of TP to DPD gene expression was significantly more frequent in endometrial cancer than in normal endometrium in any menstrual phase. DPD may act cooperatively with TP to affect cell function by maintaining the pyrimidine nucleotide pool balance in normal and malignant endometrium.

Induction of thymidine phosphorylase expression and enhancement of Furtulon sensitivity by Taxol in a human ovarian carcinoma cell line.

OBJECTIVE: Thymidine phosphorylase (TP) is one of the enzymes involved in the salvage pathways of nucleotide synthesis. The enzyme converts thymidine to thymine and 2'-deoxyribose-1-phosphate and can also metabolize the prodrug 5'-deoxy-5-fluorouridine (Furtulon) to 5-fluorouracil and 5'-deoxy-D-ribose-1-phosphate. The aim of this study was to determine whether Paclitaxel (Taxol) induces TP expression and whether increased TP expression is further sensitized to Furtulon, using a human ovarian carcinoma cell line. MATERIAL AND METHODS: KF 28, a single-cell clone of a human ovarian carcinoma cell line, was used. TP expression was assessed by RT-PCR and Western blot analysis. Cell growth was evaluated by MTT assay. RESULTS: The concentration of Taxol that inhibited the growth of cells to the level of 50% of the control growth was 4.65 +/- 0.35 nM. TP gene expression was significantly increased at 72 hours 1 nM Taxol exposure compared to the control by RT-PCR (p = 0.020). Western blot analysis confirmed that the TP protein level was elevated compared to the control at 72 hours 1 nM Taxol exposure. Cell growth did not significantly differ between the control and 72 hours 1 nM Taxol exposure groups (p = 0.917). After 48 hours treatment with Furtulon followed by 72 hours 1 nM Taxol exposure, cell growth was dose-dependently inhibited in Taxol-treated cells (p = 0.022), but not in non-Taxol-treated cells (p = 0.082). CONCLUSION: Our results show that a low concentration of Taxol is a candidate for increasing TP expression in a human ovarian carcinoma cell line, and that cells with an elevated level of TP expression can be further sensitized to Furtulon. This information might be valuable in the development of new therapeutic interventions for epithelial ovarian cancer.

Serum endostatin levels in patients with epithelial ovarian cancer.

OBJECTIVE: Endostatin, a 20-kDa C-terminal fragment of collagen XVIII, specifically inhibits endothelial proliferation and potently inhibits angiogenesis and tumor growth. The aim of this study was to assess the prognostic potentials of serum endostatin levels in epithelial ovarian cancer. MATERIALS AND METHODS: The preoperative serum levels of endostatin in 61 cases of epithelial ovarian cancer (29 serous, 13 mucinous, 13 endometrioid, 5 clear cell and 1 Brenner cell) were analyzed using a competitive enzyme immunoassay. With regard to staging, 23 cases had stage I disease, 6 had stage II disease, 28 had stage III disease and 4 had stage IV disease. RESULTS: Serum levels were compared with levels from 22 age-matched healthy volunteer blood donors. The median serum levels were 18.5 ng/ml (range, 6.3-50.3 ng/ml) in patients with epithelial ovarian cancer and 18.4 ng/ml (range, 8.4-27.0 ng/ml) in controls. No significant difference was noted between the two groups. No clinicopathological features (e.g., patients' age at diagnosis, stage of disease, histological subtype and grade) were significantly associated with serum endostatin levels. Survival data were available for all patients. Multivariate Cox regression analysis revealed that FIGO stage III-IV (p = 0.015) and serum endostatin levels (> 2 standard deviations above the control mean; 27.7 ng/ml) (p = 0.035) are independent prognostic factors. CONCLUSION: Elevated serum levels of this endogenous inhibitor of angiogenesis may be a pertinent prognostic indicator for patients with epithelial ovarian cancer. It is anticipated that this finding may aid the development of a new therapeutic strategy for epithelial ovarian cancer.

The role of thymidine phosphorylase (TP) mRNA expression in angiogenesis of prostate cancer.

BACKGROUND: Thymidine phosphorylase (TP) is associated with tumor angiogenesis. The aim of this study was to investigate the role of TP mRNA expression in the angiogenesis of prostate cancer (PC). MATERIALS AND METHODS: Prostatic tissues were obtained from 37 patients who underwent radical prostatectomy. TP and TP mRNA expression was assessed by immunohistochemistry and RT-PCR, respectively. Vascular endothelial growth factor (VEGF) expression, microvessel density (MVD) and blood flow were studied as markers of angiogenesis. RESULTS: The level of TP mRNA expression significantly correlated with the TP expression level, MVD and Gleason score in a group where TP expression was predominantly localized to stromal cells rather than malignant cells within the tumor. It also correlated with VEGF expression and blood flow, but not with clinical findings including T category and serum PSA level. CONCLUSION: This study suggests that the overexpression of TP mRNA by stromal cells within tumors plays an important role in tumor angiogenesis of PC.

Induction of thymidine phosphorylase expression by Taxol does not enhance Furtulon sensitivity in a cisplatin-resistant human ovarian carcinoma cell line.

OBJECTIVE: Thymidine phosphorylase (TP) can metabolize the prodrug 5'-deoxy-5-fluorouridine (Furtulon) to 5-fluorouracil (5-FU) and 5'-deoxy-D-ribose-1-phosphate. Furthermore, TP may enhance the toxicity of the active drug 5-FU by the transfer of 2'-deoxyribose 1-phosphate, so producing 5-fluoro-2'-deoxyuridine. This product can form 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP) through the action of thymidine kinase, and FdUMP in turn can inhibit thymidylate synthase (TS), leading to reduced thymidylate formation and subsequent inhibition of DNA synthesis. The aim of this study was to determine whether the expression of TP is associated with the sensitivity to Furtulon, using a single-cell clone of the human ovarian carcinoma cell line, KF-28, and KFr13, a cisplatin-resistant subline derived from KF28 cells. Next, if lower TP expression correlated with decreased sensitivity to Furtulon, we planned to investigate the possibility that increased TP expression induced by Paclitaxel (Taxol) exposure might increase the sensitivity to Furtulon. MATERIALS AND METHODS: Cell growth was evaluated by MTT assay. TP and TS expression were assessed by RT-PCR and Western blot analysis. RESULTS: Cell growth was significantly inhibited compared to the control at 10 (p < 0.0001), 100 (p < 0.0001) and 1000 microM (p < 0.0001) of Furtulon after 24 hours Furtulon treatment in KF28. However, cell growth was significantly inhibited only at 1000 microM (p < 0.0001), in KFr13. The expression of TP was observed only in KF28 in our PCR condition. Next, Western blot analysis confirmed that TP protein levels in KF28 were markedly elevated compared to those in KFr13. TP gene expression emerged at 72, 96 and 120 hours after 0.5 nM Taxol (about twenty percent of IC50; 2.61 +/- 0.06 nM) exposure to KFr13 in our PCR condition. The level of TP gene expression was the highest at 120 hours Taxol exposure. Similarly, Western blot analysis showed that the TP protein level of KFr13 cells 120 hours after Taxol exposure (KFr13/120 hours Taxol exposure) was elevated compared to the control. Cell growth did not significantly differ between KFr13 and KFr13/120 hours Taxol exposure cells. KFr13 and KFr13/120 hours Taxol exposure cells were incubated with 0-1000 microM Furtulon for 24 hours-168 hours. Furtulon sensitivity of KFr13/120 hours Taxol exposure cells was not found to be significantly enhanced compared to that of KFr13 cells at any of the indicated times. The level of TS gene expression assessed by RT-PCR in KFr13 and KFr13/120 hours Taxol exposure cells was significantly lower than that in KF28 cells (p < 0.001). Moreover, higher protein level expression of TS was noted in KF28 cells compared to KFr13 or KFr13/120 hours Taxol exposure cells. CONCLUSION: Our results suggest that lower expression of TP is not a critical determinant in the development of resistance to Furtulon in the cisplatin-resistant human ovarian carcinoma cell line, KFr13. The clinical relevance of these observation remains to be established.

Clinical significance of interleukin-1 receptor antagonist in patients with cervical carcinoma.

OBJECTIVE: Our purpose was to determine the clinical significance of interleukin-1 receptor antagonist (IL-1ra), which is an endogenous inhibitor cytokine of IL-1, in patients with cervical carcinoma. METHODS: Tissue IL-1ra expression and serum IL-1ra level were examined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and immunohistochemistry in normal controls and patients with cervical carcinoma. RESULTS: Tissue IL-1ra protein level by ELISA was significantly higher in squamous cell carcinoma (n = 9) than in the normal cervix (n = 7) and adenocarcinoma (n = 3). Western blotting confirmed the main presence of intracellular IL-1ra type 1 in squamous cell carcinoma. Immunohistochemistry demonstrated significant IL-1ra expression only in tumor cells of squamous cell carcinoma. Elevation of serum IL-1ra level was found in patients with squamous cell carcinoma (n = 38) compared to normal women (n = 13), but not in patients with adenocarcinoma (n = 9). Although serum IL-1ra level did not correlate with clinical stage or any other tumor marker, high serum IL-1ra level was associated with pelvic lymph node metastasis and poor prognosis in patients with squamous cell carcinoma. On the other hand, these results were not obtained in patients with cervical adenocarcinoma. CONCLUSION: IL-1ra may play important roles in local and general malignant behaviors in patients with cervical squamous cell carcinoma, and measurement of serum IL-1ra level may be useful in predicting patient survival.

Allelotypes as potential prognostic markers in ovarian carcinoma treated with cisplatin-based chemotherapy.

We analyzed a series of ovarian carcinomas from patients treated with cisplatin-based chemotherapy for loss of heterozygosity (LOH) to determine the relationship between microsatellite alteration and prognosis. Patients with tumors that had lost alleles at loci 3p14-21, 12qter, or 17p13.3 showed significantly reduced survival compared to patients with tumors that retained both alleles at those loci. The 5-year mortality rates for patients exhibiting allele loss and patients with allele retention were 50 and 41%, respectively, for the 3p14-21 locus (P=0.0023); 57 and 24%, respectively, for 12qter (P=0.0256); and 55 and 40%, respectively, for 17p13.3 (P=0.0489). A statistically significant difference was also observed with respect to fractional allelic loss (FAL), which was a significant indicator for disease recurrence (P=0.0227). The prognosis of patients with high FAL values were significantly worse compared to those with low FAL values (P=0.0306). Our results suggested that LOH at loci 3p14-21, 12qter, or 17p13.3 was a significant predictor of poor survival in ovarian carcinomas treated with cisplatin-based chemotherapy. Furthermore, the association of FAL value with therapy response indicated that ovarian carcinomas with high levels of chromosomal alteration may be more resistant to this type of chemotherapy.

Prognostic value of overexpression of p53 in human ovarian carcinoma patients receiving cisplatin.

A major obstacle to the treatment of ovarian carcinoma is intrinsic/acquired resistance to cisplatin-based chemotherapy. The clinical significance of p53 overexpression in patients with ovarian carcinoma is still controversial. The aim of this study was to investigate the independent prognostic significance of p53 overexpression in patients with ovarian carcinoma who are treated with cisplatin. We retrospectively examined the overexpression of p53 in primary ovarian carcinoma, and its association with chemotherapeutic efficacy. One hundred and thirty four ovarian carcinomas were surgically removed from patients who received adjuvant cisplatin-based chemotherapy. Immunohistochemical analysis of p53 was performed using a DO7 antibody against the p53 protein in 134 ovarian carcinomas. The significance of p53 in the prognosis of patients with ovarian carcinomas was also examined by a survival analysis of mortality follow-up data covering the period from 1988 to 2001. Thirty-three tumors (25%) exhibited p53 overexpression. Overexpression of p53 in grade 2/grade 3 tumors was significantly higher than that seen in grade 1 tumors (P=0.0088, 0.0229). Patients with tumors who also showed overexpression of p53 had a significantly inferior response to chemotherapy compared with the patients with p53-negative tumors (P=0.04). Cox regression analysis revealed that p53 overexpression was prognostic for poor disease outcome after adjustment for FIGO stage, grade and residual tumor. These findings suggest that overexpression of p53 in ovarian carcinoma is associated with unfavorable clinical outcome in patients treated with cisplatin-based chemotherapy. Therefore, detection of p53 overexpression using the DO7 antibody may be considered as a predictive marker of chemoresistance for cisplatin in patients with ovarian carcinoma.