竹井 裕二

Vasohibin-1 (VASH1) is a negative feedback regulator of angiogenesis, the first to be discovered, and was identified in vascular endothelial growth factor (VEGF)-stimulated vascular endothelial cells. Vasohibin-1 inhibits abnormal vascularization induced by various angiogenic factors including fibroblast growth factor and platelet-derived growth factor (PDGF), in addition to VEGF. By focusing on this characteristic of VASH1, we investigated the antitumor effects of VASH1 expression on ovarian cancer cells that produce different angiogenic factors. By using a high VEGF-producing ovarian cancer cell line, SHIN-3, and a high PDGF-producing ovarian cancer cell line, KOC-2S, the cells were transfected with either a VEGF antagonist, soluble VEGF receptor-1 (sVEGFR-1, or sFlt-1), or VASH1 genes to establish their respective cellular expression. The characteristics of these transfectants were compared with controls. We previously reported that the expression of sFlt-1 inhibited tumor vascularization and growth of high VEGF-producing ovarian cancer cells, reduced peritoneal dissemination and ascites development, and prolonged the survival time of the host. However, in the current study, the expression of sFlt-1 had no such effect on the high PDGF-producing ovarian cancer cells used here, whereas VASH1 expression inhibited tumor vascularization and growth, not only in high VEGF-producing cells, but also in high PDGF-producing cells, reduced their peritoneal dissemination and ascites, and prolonged the survival time of the host. These results suggest that VASH1 is an effective treatment for ovarian cancer cells that produce different angiogenic factors.

術前にadenocarcinoma in situ(AIS)の診断で単純子宮全摘出術をしたところ、術後の病理診断で子宮頸部腺癌IB1期であった2症例を経験した。1例目は42歳で、子宮頸部狙い組織診でAISと診断した。一期的な治療の希望が強く、単純子宮全摘出術をしたところ子宮頸部腺癌IB1期であった。2例目は53歳で、子宮頸部狙い組織診と円錐切除術の病理組織結果からAIS(断端陽性)と診断した。追加治療として単純子宮全摘出術をしたところ子宮頸部腺癌IB1期であった。生検や円錐切除術で診断したAISは過少診断となる可能性があることを再認識した。(著者抄録)

子宮結核は特徴的な症状がなく、検診などで偶発的に発見されることがある。今回、子宮留膿症患者に施行した子宮内膜細胞診が、子宮結核の診断の契機となった症例を経験した。結核性子宮内膜炎の細胞像は、ラングハンス型巨細胞、類上皮細胞の出現を特徴とし、これらの所見から結核性病変を強く疑うことができた。子宮内膜細胞診は悪性疾患の除外のみならず、子宮結核の診断の契機として有用であった。子宮内膜細胞診所見で結核を疑うことは早期の治療介入につながると思われる。(著者抄録)

Uterine clear cell adenocarcinoma (UCCA) is rare and resistant to treatment. We report a UCCA patient who responded to radiotherapy on each relapse. The first relapse was detected in the vaginal wall after the first course of postoperative adjuvant chemotherapy. Radiotherapy was conducted. Recurrent tumors were detected in the left lung after 5 months and in the right lung after 8 months. Partial resection of the lungs was performed. After 5 months, relapse was detected in the left pulmonary apex. Stereotactic radiotherapy was conducted. After 7 months, relapse was detected in the left pulmonary apex outside the irradiation field, and stereotactic radiotherapy was performed. During the subsequent 36-month follow-up, there has been no relapse. Although UCCA is resistant to treatment, radiotherapy is effective in some cases, as demonstrated in this patient. Even when relapse is repeated, radiotherapy may be considered as a treatment option if the recurrent focus is localized.

Vasohibin-1 (VASH1) is expressed in vascular endothelial cells stimulated by several angiogenic growth factors and displays autocrine activity to regulate angiogenesis via a negative feedback mechanism. In this study, we investigated the effect of VASH1 on ovarian cancer progression using VASH1-expressing ovarian cancer cells in vitro and in vivo. The growth ability of ovarian cancer cells engineered to express the VASH1 gene remained unchanged in vitro. However, we showed that VASH1 secretion by tumor cells inhibited the growth of human umbilical vein endothelial cells. Further, animal experiments showed that VASH1 expression inhibited tumor angiogenesis and growth. In a murine model of peritoneal dissemination of ovarian cancer cells, VASH1 inhibited peritoneal dissemination and ascites, resulting in significantly prolonged survival in mice. This indicates that VASH1 exerts an antitumor effect on ovarian cancer by inhibiting angiogenesis in the tumor environment. These findings suggest that a novel therapy based on VASH1 could be a useful therapeutic strategy for ovarian cancer.

PURPOSE: We previously reported that the concept of "platinum sensitivity" could be applied to recurrent endometrial cancer. We conducted an ancillary analysis to determine an appropriate second-line regimen for patients who received a platinum agent as first-line chemotherapy. METHODS: We extracted and reanalyzed data of patients treated with doxorubicin and cisplatin (AP), paclitaxel and carboplatin (TC), or docetaxel and carboplatin (DC) as first- and second-line chemotherapies from the SGSG012/GOTIC004/Intergroup study. RESULTS: We identified 216 patients: 38 received AP as first-line chemotherapy, of which 36 received TC or DC (Tax-C) as second-line chemotherapy; and 178 received Tax-C as first-line chemotherapy, of which 51 received AP and 127 received Tax-C as second-line chemotherapy. Median progression-free survival (PFS) and overall survival (OS) after second-line chemotherapy decreased in the order of Tax-C followed by Tax-C (10 and 48 months, respectively), AP followed by Tax-C (9 and 23 months, respectively), and Tax-C followed by AP (3 and 12 months, respectively). Median PFS and OS after second-line chemotherapy for platinum-resistant patients receiving Tax-C as first-line chemotherapy were longer in Tax-C than in AP (7 and 23 vs. 3 and 10 months, respectively) as second-line chemotherapy [hazard ratio (HR) 3.255, 95 % confidence interval (CI) 1.908-5.555, p < 0.0001; HR 3.179, 95 % CI 1.835-5.507, p < 0.0001, respectively]. Median PFS and OS after second-line chemotherapy for platinum-sensitive patients receiving Tax-C as first-line chemotherapy were almost equivalent to those receiving Tax-C or AP as second-line chemotherapy. CONCLUSIONS: For platinum-resistant recurrent endometrial cancer patients, Tax-C may be preferred over AP as second-line chemotherapy.

Primary gestational choriocarcinoma is commonly present in the uterus in cases of atypical genital bleeding. Symptoms similar to those of an ectopic pregnancy develop when an extra-uterine lesion is present in the abdominal cavity, and lesions have been detected in the ovaries and fallopian tubes in a number of cases. In the present study, we describe a patient with choriocarcinoma that metastasized to the uterine serosa and caused symptoms similar to those of an ectopic pregnancy. The patient was a 30-year-old female who presented to our hospital with atypical genital bleeding and a positive pregnancy test 3 months after missed abortion at 10 weeks of gestation. Transvaginal ultrasonography revealed the absence of a gestational sac in or outside the uterus, and intra-abdominal bleeding was noted. An ectopic pregnancy was suspected based on these findings, and emergency laparotomy was performed. A hemorrhagic mass was present on the uterine serosa, and was subsequently resected. Trophoblastic disease was suspected following histopathological examination, for which intra-uterine curettage was performed and choriocarcinoma was diagnosed. Lung metastasis was detected on computed tomography, and a high serum human chorionic gonadotropin (hCG) level persisted following surgery. The lesion disappeared following five cycles of methotrexate+etoposide+actinomycin D therapy, which was performed as postoperative chemotherapy, and the patient's serum hCG level decreased to below the detection limit. In this case of choriocarcinoma, the primary lesion was present in the uterus and had metastasized to the uterine serosa, which is a very rare metastatic site. This uterine serosal metastatic lesion bled and caused symptoms similar to those of an ectopic pregnancy. Certain patients who undergo surgery for a suspected peritoneal pregnancy may have gestational choriocarcinoma, similar to this case.

卵巣成熟嚢胞性奇形腫の悪性転化はほとんどが扁平上皮癌であり、腺癌への悪性転化は稀である。また、腹膜偽粘液腫の多くは虫垂由来で、卵巣原発例は少数であるとされている。今回、我々は卵巣成熟嚢胞性奇形腫から腹膜偽粘液腫を伴う粘液性腺癌へ悪性転化を示した1例を経験した。本症例から、腹膜偽粘液腫は卵巣成熟嚢胞性奇形腫から発生し得ることを学んだ。腹膜偽粘液腫の原発臓器の特定には、虫垂切除および卵巣成熟嚢胞性奇形腫の有無についての詳細な病理学的検索が重要である。(著者抄録)

Lobular endocervical glandular hyperplasia (LEGH) is histologically similar to minimal deviation adenocarcinoma (MDA), but classified as a benign disease. Although MDA often develops in Peutz-Jeghers syndrome (PJS) patients, there have been only a few reports on PJS with LEGH. We report a PJS patient who was diagnosed with LEGH by conization and delivered a baby 42 months later. She was referred to our department for multicystic lesions in the uterine cervix at 26 years old. Diagnostic conization was performed, and the histopathological diagnosis was LEGH. As the possibility of MDA could not be ruled out because of concomitant PJS, hysterectomy was considered. However, course observation was selected because the patient strongly wished to preserve fertility. She delivered a baby at 30 years old. The finding that PJS patients may be complicated by LEGH is very important. Loss of fertility by over-treatment should be avoided if patients desire its preservation.

BACKGROUND: In Japan, the cervical cancer screening rate is extremely low. Towards improving the cervical cancer screening rate, encouraging eligible people to make an informed choice, which is a decision-making process that relies on beliefs informed by adequate information about the possible benefits and risks of screening, has attracted increased attention in the public health domain. However, there is concern that providing information on possible risks of screening might prevent deter from participating. METHODS: In total, 1,912 women aged 20-39 years who had not participated in screening in the fiscal year were selected from a Japanese urban community setting. Participants were randomly divided into 3 groups. Group A received a printed reminder with information about the possible benefits of screening, group B received a printed reminder with information about possible benefits and risks, and group C received a printed reminder with simple information only (control group). RESULTS: Out of 1,912 participants, 169 (8.8%) participated in cervical cancer screening. In the intervention groups, 137 (10.9%) participated in cervical cancer screening, compared to only 32 (4.9%) of the control group (p < 0.001). In addition, logistic regression analysis revealed that there was no significant difference in screening rate between group A and group B (p = 0.372). CONCLUSIONS: Providing information on the possible risks of screening may not prevent people from taking part in cervical cancer screening among a Japanese non-adherent population.

BACKGROUND: The aim of this study was to investigate the impact of the histological findings on the treatment of malignant ovarian tumors in pregnant women. METHODS: This is a retrospective study of 41 patients diagnosed and treated for ovarian malignancy during pregnancy between 1985 and 2010. RESULTS: The median age of the study group was 30 years old, ranging from 20 to 41. Thirty-eight (92 %) patients were diagnosed with stage I, and one (2 %) with each of stages II, III, and IV. Twenty-five (61 %) patients had borderline malignancy, 8 (20 %) were diagnosed with epithelial ovarian cancer, 7 (17 %) with germ cell tumor, and one with sex cord stromal tumor. All patients received primary surgery; 7 (17 %) patients had cystectomy, 32 (78 %) had unilateral salpingo-oophorectomy, and 3 (7 %) underwent hysterectomy with bilateral salpingo-oophorectomy. Thirty-one (76 %) patients delivered live newborns; 21 had borderline tumor (84 %), 2 had ovarian cancers (25 %), and 8 had non-epithelial tumor (100 %). Six cases were terminated in order to perform the standard treatment for ovarian malignancy and 2 cases aborted spontaneously. CONCLUSION: In pregnant women, ovarian cancer is exceptionally less frequent compared with non-pregnant women, i.e. age-matched, statistically-corrected controls based on the Japanese annual report [8/33 (24 %) vs. control (60 %); ovarian cancer/(ovarian cancer + borderline tumor), P = 0.001]. The pregnant women with ovarian cancer chose to prioritize treatment of ovarian cancer at the sacrifice of their babies while those with borderline tumor or non-epithelial tumor were able to successfully deliver live newborns.

Small cell carcinoma of the cervix (SCCC) is a rare histological entity of uterine cervical cancer. Compared with other common histological types, squamous cell carcinoma or adenocarcinoma, the outcome of SCCC is poor because of the high incidence of nodal or distant metastasis even with early stage. In this review, current consensus of epidemiology, pathology, and initial treatment for SCCC will be discussed.

OBJECTIVE: Endometrial cancer often coexists with uterine adenomyosis. However, little is known about the clinical characteristics of these cases. Thus, cases of endometrial cancer occurring with and without uterine adenomyosis were compared, and the influences of uterine adenomyosis on the clinical progress of endometrial cancer were examined. MATERIALS AND METHODS: Of endometrial cancer patients who underwent hysterectomies in our facility from 2002 to 2011, we included only endometrioid adenocarcinoma patients in our study. The patients were divided into 2 groups, adenomyosis group and nonadenomyosis group, according to the presence/absence of uterine adenomyosis. Patient characteristics, stage, histopathological grade, muscle invasion, recurrence, and mortality were retrospectively compared and examined. RESULTS: There were 362 cases of endometrioid adenocarcinoma of the uterine body, of which 121 (33.4%) and 241 cases (66.6%) were in the adenomyosis and nonadenomyosis group, respectively. There were no significant differences with respect to the disease stages or ratios of the histopathological grade between the 2 groups. In the adenomyosis group/nonadenomyosis group, 5-year progression-free survival for International Federation of Gynecology and Obstetrics (FIGO) stages I and II was 89.9%/93.7% and that for stages III and IV was 70.6%/62.0%; the 5-year overall survival was 100%/95.9% for FIGO stages I and II, and 88.0%/73.5% for stages III and IV. There were no significant between-group differences for either progression-free survival or overall survival. When limiting the results to only FIGO stage I endometrioid adenocarcinoma, despite no grade variance between the 2 groups, a significant difference was observed in the ratios of outer-half muscle invasion between the adenomyosis and nonadenomyosis groups (19.5% [17/87] vs 10.1% [16/158], P < 0.05); however, the prognosis was similar in the 2 groups. CONCLUSIONS: Uterine adenomyosis is associated with deep myometrial invasion in stage I endometrioid adenocarcinoma; however, it did not affect the recurrence or mortality rates.

2011-2012年の2年間に自治医科大学附属病院婦人科にて確定診断された婦人科悪性腫瘍(卵巣がん77例、子宮頸がん93例、子宮体がん113例)の発見契機を後方視的に検討した。各々の婦人科がんには特徴的な発見契機があり、これらを詳細に検討することは、早期発見の手掛かりとなると考えられる。特に卵巣がんにおいては、約半数の症例が婦人科以外の科を初診しており、他科との連携が重要であることが浮き彫りとなった。このような情報は婦人科から他科へ発信すべきものと考えられた。(著者抄録)

OBJECTIVES: While radiation therapy is administered as a palliative treatment for recurrent ovarian cancer, it remains unclear whether it improves the prognosis. METHODS: The effects and adverse events of radiation therapy for patients with recurrent epithelial ovarian cancer were investigated using medical records. RESULTS: Herein, 46 subjects comprising 33 patients whose recurrent lesions were contained within the irradiation field (therapeutic radiation group; TRG) and 13 patients with some recurrent lesions outside the irradiation field (palliative radiation group; PRG) were included. The TRG achieved a response rate (RR) of 66%, a disease control rate (DCR) of 100%, a progression-free survival (PFS) of 10 months, and an overall survival (OS) of 20 months. The PFS after radiation therapy was significantly longer than that following chemotherapy received just before radiation therapy. The PFS of patients with recurrent intrapelvic lesions was longer than that of patients with some extrapelvic recurrence. There was no significant association between PFS after radiation therapy and the duration from the previous chemotherapy or histological type. The RR, DCR, PFS, and OS of the PRG were 30 and 90% and 2 and 6 months, respectively. Serious adverse events were rare. CONCLUSIONS: Radiation therapy is a potential option for chemotherapy-resistant, localized recurrent ovarian cancer. © 2014 S. Karger AG, Basel.

A leiomyoma rarely causes disseminated intravascular coagulopathy (DIC). In the present report, we describe a case of DIC caused by leiomyoma. A 36-year-old nulliparous woman presented with hypermenorrhea and a lower abdominal mass. On magnetic resonance imaging, we detected a 14 cm uterine tumor, which was suspected to be a sarcoma. Blood tests at the preoperative examination indicated platelet count of 9.6 × 10(4)/μL, fibrin degradation product level of 107.1 μg/mL (normal value, 0-5.0 μg/mL), and fibrinogen level of 54 mg/dL (normal value, 129-271 mg/dL). Based on these findings, we diagnosed the patient with DIC. The patient was treated with nafamostat mesilate and fresh frozen plasma, but the DIC did not show any improvement. Subsequently, a hysterectomy was performed, after which the DIC improved. Clinicopathological findings indicated the presence of a leiomyoma with multiple vessels containing thromboemboli, and suggested that the DIC was caused by the leiomyoma. Therefore, it is essential to consider that that a benign leiomyoma may be a cause of DIC.

The purpose of this study was to examine the effect of various community-based interventions in support of HPV vaccination implemented by cities and towns within Tochigi prefecture, Japan with a view to identifying useful indicators which might guide future interventions to improve HPV vaccination coverage in the prefecture. A postal questionnaire survey of all 27 local governments in Tochigi Prefecture was conducted in December 2010. All 27 responded, and 22 provided the exact numbers of the targeted and vaccinated populations of 13- to 15-year-old girls from April to December 2010. The local governments also answered questions on the type of interventions implemented including public subsidies, school-based programs, direct mail, free tickets and recalls. Local governments that conducted a school-based vaccination program reported 96.8% coverage for the 1st dose, 96.2% for the 2nd dose, and 91.2% for the 3rd dose. Those that provided subsidies without school-based programs reported a wide range of vaccination rates: 45.7%-95.0% for the 1st dose, 41.1%-93.7% for the 2nd dose and 3.1%-90.1% for the 3rd dose. Among this group, the combination of a free ticket, direct mail and recall was most effective, with 95.0% coverage for the 1st dose, 93.7% for the 2nd dose, and 90.1% for the 3rd dose. The governments that did not offer a subsidy had the lowest vaccination coverage, with 0.8%-1.4% for the 1st dose, 0.0%-0.8% for the 2nd dose, and 0.1%-0.1% for the 3rd dose. The results of this survey indicate that school-based vaccinations and public subsidies are the most effective method to improve HPV vaccination coverage; however, the combination of a free ticket, direct mail, and recalls with public subsidies are also important measures in increasing the vaccination rate. These data may afford important indicators for the successful implementation of future HPV vaccination programs.

Vasohibin-2 (VASH2) is a homolog of vasohibin-1 and exhibits pro-angiogenic activity. We recently reported that VASH2 is expressed in certain ovarian cancers and promotes tumor growth through angiogenesis. To further demonstrate the effectiveness of molecular targeting of VASH2 for anticancer treatment, we applied in vivo delivery of siRNA targeting VASH2 (siVASH2) using atelocollagen to a xenograft model of ovarian cancer. We inoculated mice s.c. with DISS and SKOV-3, two representative human ovarian serous adenocarcinoma cell lines. When tumors were measurable, we initiated treatment with control or siVASH2 mixed with atelocollagen, which enveloped the whole tumor. Treatment with siVASH2 significantly inhibited s.c. tumor growth by abrogating tumor angiogenesis. We confirmed that expression of VASH2 mRNA in the tumor was downregulated by siVASH2 treatment. In addition, the siVASH2-treated tumor contained more blood vessels covered with pericytes, indicating that knockdown of VASH2 contributes to the normalization of tumor blood vessels. Based on these results, VASH2 may be a promising molecular target for ovarian cancer treatment.

Lymph node metastasis is the most important prognostic factor of endometrial cancer. However, effective therapy has not been established against lymph node metastasis. In this study, we explored the efficacy of gene therapy targeting lymph node metastasis of endometrial cancer by suppressing the action of vascular endothelial growth factor (VEGF)-C through soluble VEGF receptor-3 (sVEGFR-3) expression. For this purpose, we first conducted a model experiment by introducing sVEGFR-3 cDNA into an endometrial cancer cell line HEC1A and established HEC1A/sVEGFR-3 cell line with high sVEGFR-3 expression. The conditioned medium of HEC1A/sVEGFR-3 cells inhibited lymphatic endothelial cell growth in vitro, and sVEGFR-3 expression in HEC1A cells suppressed in vivo lymph node and lung metastases without inhibiting the growth of a subcutaneously inoculated tumor. To validate the therapeutic efficacy, adeno-associated virus vectors encoding sVEGFR-3 were injected into the skeletal muscle of mice with lymph node metastasis. Lymph node and lung metastases of HEC1A cells were completely suppressed by the muscle-mediated expression of sVEGFR-3 using adeno-associated virus vectors. These results suggest the possibility of gene therapy against lymph node and lung metastases of endometrial cancer by using muscle-mediated expression of sVEGFR-3.

This study examined the role of the immunosuppressive enzyme indoleamine-2,3-dioxygenase (IDO) in cervical cancer progression and the possible use of this enzyme for cervical cancer therapy. We analyzed IDO protein expression in 9 cervical cancer cell lines (SKG-I, -II, -IIIa, -IIIb, SiHa, CaSki, BOKU, HCS-2 and ME-180) stimulated with interferon-γ. IDO expression was observed in all cell lines except for SKG-IIIb. We transfected the human cervical cancer cell line CaSki that constitutively expresses IDO with a short hairpin RNA vector targeting IDO, and established an IDO-downregulated cell line to determine whether inhibition of IDO mediates cervical cancer progression. IDO downregulation suppressed tumor growth in vivo, without influencing cancer cell growth in vitro. Moreover, IDO downregulation enhanced the sensitivity of cervical cancer cells to natural killer (NK) cells in vitro and promoted NK cell accumulation in the tumor stroma in vivo. These findings indicate that downregulation of IDO controls cervical cancer progression by activating NK cells, suggesting IDO as a potential therapy for cervical cancer.

This study retrospectively compared nedaplatin and irinotecan hydrochloride (NDP/CPT) combination therapy with cisplatin and irinotecan hydrochloride therapy (CDDP/CPT) for efficacy and adverse events in the treatment of clear cell adenocarcinoma of the ovary (CCC) and recurrent ovarian carcinoma. A total of 115 patients were included in the present study. NDP/CPT was administered intravenously every 4 weeks (NDP, 60 mg/m(2) on day 1; CPT, 50 mg/m(2) on days 1, 8 and 15). CDDP/CPT was also administered intravenously (CDDP, 60 mg/m(2) on day 1; CPT, 60 mg/m(2) on days 1, 8 and 15). Patients with primary CCC were treated with NDP/CPT in 29 cases and CDDP/CPT in 20 cases. Patients with recurrent ovarian carcinoma were treated with NDP/CPT and CDDP/CPT in 33 cases each. No significant difference was observed in the 5-year overall survival (OS)/progression-free survival (PFS) of patients with primary CCC, with the exception of those patients with stages Ia and Ic(b) who underwent NDP/CPT and CDDP/CPT treatments (OS: 58%, PFS: 40% and OS: 53% and PFS: 47%, respectively). No significant differences were found in the response rates to NDP/CPT and CDDP/CPT in patients with recurrent ovarian carcinoma (27 and 18%, respectively). Similarly, there were no significant differences in the 5-year OS and PFS of patients with recurrent ovarian carcinoma treated with NDP/CPT or CDDP/CPT (OS: 15%, PFS: 3% and OS: 18%, PFS: 6%, respectively). In terms of the hematological toxicity of grade 3 or above and non-hematological toxicity of grade 2 or above in patients treated with NDP/CPT and CDDP/CPT, respectively, neutropenia was 23 and 56%; anemia, 1, and 20%; thrombocytopenia, 0 and 5%; nausea, 20 and 52%; diarrhea, 14 and 25%; and fever, 2 and 11%. Accordingly, NDP/CPT indicated mild toxicity, and was therefore equally effective and less toxic than CDDP/CPT in the treatment of primary CCC and recurrent ovarian carcinoma.

PURPOSE: Although the pharmacokinetic mechanism of nedaplatin (NDP) is similar to carboplatin, the dose of NDP is typically determined by body surface area and not by the area under the curve (AUC). We conducted a phase I study to determine the AUC-calculated optimal dosage of NDP used in combination chemotherapy with irinotecan (CPT-11) for gynecologic malignancies. METHODS: A total of 15 patients who were to undergo combination chemotherapy consisting of NDP and CPT-11 were enrolled in this study. The dose of CPT-11 was administered at a fixed dose of 60 mg/m(2) and that of NDP was gradually increased from 8 to 12 μg h/mL (AUC). The individual dose of NDP was calculated based on creatinine clearance of the patient according following formula: Dose(NDP) = AUC × CL(NDP), where CL(NDP) = 0.0738 × creatinine clearance + 4.47 (Ishibashi's formula). RESULTS: One patient had dose-limiting toxicity (DLT) at level 1, and two patients suffered DLT at level 3. The dosage of NDP at AUC 12 was determined to be the maximum tolerated dose in combination chemotherapy with CPT-11, even though only two of the six patients showed DLT at level 3. CONCLUSIONS: The recommended dosage of NDP calculated by AUC with Ishibashi's formula was set to AUC 10 in combination chemotherapy with CPT-11.

The purpose of this study was to explore the possibility of targeted molecular therapy with anti-epidermal growth factor receptor (anti-EGFR) antibody (cetuximab) for the treatment of mucinous ovarian carcinoma. We analyzed EGFR protein expression and KRAS gene mutations in 5 mucinous ovarian carcinoma cell lines RMUG-L, RMUG-S, MN-1, OMC-1 and MCAS and evaluated the in vitro and in vivo effects of cetuximab on each. EGFR expression was observed in all cell lines except for MN-1 cells, and a KRAS gene mutation at codon 12 was detected only in the MCAS cell line. Cetuximab inhibited RMUG-L and OMC-1 cell growth in vitro and completely blocked RMUG-L tumor growth in vivo. On the other hand, cetuximab did not affect MCAS cell growth in vitro and only partially reduced the MCAS tumor growth in vivo. These results suggest the possibility of targeted molecular therapy with cetuximab for mucinous ovarian carcinoma cells lacking a KRAS gene mutation.

The purpose of this study was to present the results of fertility-sparing treatment using medroxyprogesterone acetate (MPA) for endometrial carcinoma (EC), and to clarify patient characteristics by investigating patient background factors. A total of 59 patients with EC, who received MPA as fertility-sparing therapy at two institutions over a 21-year period between 1987 and 2008, were studied retrospectively. Patients were administered oral MPA at 400-600 mg/day for 16-24 weeks as long as they responded. Endometrial tissue was assessed twice, at 8-12 weeks (during treatment) and shortly after treatment. The overall complete response (CR) rate was 71%. A total of 22 (52%) of 42 responders later developed relapse. A total of 19 cases became pregnant, and 25 infants were born. Eighty percent of recurrences occurred within 2 years. For stages la and Ib-IIa (FIGO, 1988), initial CR rates were 80.0 and 42.9%, respectively (p&lt;0.01), demonstrating a significant difference. Total hysterectomy was performed for 26 patients (44%) due to recurrence or failure to respond to the initial treatment. Among these 26 patients, postoperative stages were more advanced in 10 patients (38%). The grade advanced (became more poorly differentiated) postoperatively in 2 patients (8%). Premenopausal females with EC can be treated successfully with MPA, however patients should be informed of the risks and limitations of this conservative treatment.

This study examined the role of the immuno-suppressive enzyme indoleamine-2,3-dioxygenase (IDO) in ovarian cancer progression, and the possible application of this enzyme as a target for ovarian cancer therapy. We transfected a short hairpin RNA vector targeting IDO into the human ovarian cancer cell line SKOV-3, that constitutively expresses IDO and established an IDO downregulated cell line (SKOV-3/shIDO) to determine whether inhibition of IDO mediates the progression of ovarian cancer. IDO downregulation suppressed tumor growth and peritoneal dissemination in vivo, without influencing cancer cell growth. Moreover, IDO downregulation enhanced the sensitivity of cancer cells to natural killer (NK) cells in vitro, and promoted NK cell accumulation in the tumor stroma in vivo. These findings indicate that downregulation of IDO controls ovarian cancer progression by activating NK cells, suggesting IDO targeting as a potential therapy for ovarian cancer.

PURPOSE: Nedaplatin (NDP), a platinum derivative, has been developed to reduce nephrotoxicity and gastrointestinal toxicity of cisplatin. The pharmacokinetic profile of NDP is similar to that of carboplatin (CBDCA). The optimal dosing for CBDCA is determined by the area under the curve (AUC) using Calvert's formula. However, the administration dose of nedaplatin (NDP) is determined based on the body surface area in clinical treatment. Ishibashi et al. reported a formula for predicting NDP clearance based on renal function like Calvert's formula for CBDCA. We conducted the present study to evaluate the Ishibashi's formula. METHODS: A total of 22 patients with cervical or ovarian cancer, who underwent chemotherapy consisting of NDP and irinotecan (CPT-11), were examined in this study. Blood samples were collected at 0, 1, 2, 4, and 6 h after the end of infusion of NDP (48-80 mg/m(2)), and free platinum concentrations were measured. Observed AUCs were compared with predicted AUCs, which were calculated by the Ishibashi's formula. In addition, the relative reduction in platelets (PLTs) was assessed as a parameter of adverse effects. RESULTS: The observed AUC of NDP ranged from 4 to 14 (μg h(-1) ml(-1)) with large variation. The predicted AUC based on renal function was correlated with the observed AUC. There was a relationship between observed AUC and the decrease in PLTs. CONCLUSIONS: Ishibashi's formula would be predictable and useful for estimating the individual dose of NDP.

Controlling lymph node metastasis is currently a key issue in cancer therapy. Lymph node metastasis is one of the most important prognostic factors in various types of cancers, including endometrial cancer. Vascular endothelial growth factor-C (VEGF-C) plays a crucial role in lymphangiogenesis, and is implicated to play an important role in lymph node metastasis. To evaluate the role of VEGF-C in lymph node metastasis, we developed an animal model by using an endometrial cancer cell line, HEC1A. This cell line is not invasive by nature and secretes moderate amounts of VEGF-C; intrauterine injection of HEC1A cells into Balb/c nude mice resulted in uterine cancer with lymph node metastasis after 8 weeks. To analyze the contribution of VEGF-C to lymph node metastasis, its corresponding gene was stably introduced into HEC1A cells (HEC1A/VEGF-C), which then produced more than 10 times the amount of VEGF-C. The number of lymph node metastases was significantly higher in HEC1A/VEGF-C cells than in HEC1A cells (3.2 vs 1.1 nodes/animal, respectively). Augmented lymphangiogenesis was observed within tumors when HEC1A/VEGF-C cells were inoculated. These results indicate that VEGF-C plays a critical role in lymph node metastasis, in addition to serving as a platform to test the efficacy of various therapeutic modalities against lymph node metastasis.

The purpose of this study was to clarify the relationship between ovarian cancer peritoneal dissemination and indoleamine 2,3-dioxygenase (IDO) expression, and to explore the possibility of IDO-targeting molecular therapy for ovarian cancer. We transfected an IDO expression vector into the IDO-non-expressing human ovarian cancer cell line OMC-1, and established an IDO-expressing cell line (OMC-1/IDO) to examine the relationship between IDO expression and cancer cell growth in vitro and in vivo. IDO expression did not influence cancer cell growth and invasion in vitro, but promoted tumor growth and peritoneal dissemination in vivo. Immunostaining showed that IDO expression inhibited natural killer (NK) cell accumulation in tumors and promoted tumor angiogenesis. In addition, the oral administration of the IDO inhibitor 1-methly-tryptophan inhibited the growth of OMC-1/IDO-derived subcutaneous tumors in mice. These findings indicate that IDO promotes the peritoneal dissemination of ovarian cancer by inhibiting NK cell accumulation in tumors and promoting angiogenesis, supporting the applicability of IDO-targeting molecular therapy in ovarian cancer.

The first aim of our study was to develop a pregnant mouse model for preeclampsia using adenoviral vector containing mouse full-length soluble fms-like tyrosine kinase 1 (sFlt-1) but not truncated sFlt-1. The second aim was to evaluate effects of recombinant mouse (rm) vascular endothelial growth factor (VEGF) and rm placental growth factor (PlGF) on a preeclampsia model induced by adenoviral vector containing mouse full-length sFlt-1. We injected adenoviral vector containing mouse full-length sFlt-1 on day 8.5 or 9.5 of gestation into pregnant Institute of Cancer Research mice, resulting in hypertension, proteinuria, and similar glomerular histological changes as those seen in human preeclamptic women with glomerular endotheliosis on day 16.5 or 17.5 of gestation. The preeclampsia models were treated with 100 microg/kg of rmVEGF164 (n=5), 100 microg/kg of rmPlGF-2 (n=5), or vehicle (n=7) twice a day for 2 days IP. The rmVEGF164 treatment significantly decreased the mean blood pressure on day 16.5 or 17.5 of gestation compared with the vehicle treatment (85+/-4 versus 97+/-2 mm Hg; P=0.018). The rmPlGF-2 treatment also significantly decreased the mean blood pressure on day 16.5 or 17.5 of gestation compared with the vehicle treatment (86+/-3 versus 97+/-2 mm Hg; P=0.018). However, proteinuria was not affected by either rmVEGF164 or rmPlGF-2. In conclusion, we, for the first time, created a mouse preeclampsia model using mouse full-length sFlt-1. VEGF and PlGF may be promising for ameliorating hypertension in women with preeclampsia. Additional study of PlGF as a potential drug for preeclampsia is warranted.

The purpose of this study was to explore the possibility of molecular-targeted therapy with anti-epidermal growth factor receptor (EGFR) antibody (cetuximab) for endometrial cancer to develop a new treatment for advanced endometrial cancer. We analyzed EGFR protein expression and gene mutations in the human endometrial cancer cell line HEC1A, and evaluated the in vitro and in vivo effects of cetuximab on HEC1A. EGFR expression was observed in HEC1A cells, but no mutations in the EGFR gene were detected. Cetuximab inhibited HEC1A cell growth and invasion and VEGF-A production in vitro, and HECIA cell tumor growth, its peritoneal dissemination with ascites, and lymph node and lung metastasis in vivo. In addition, the antibody prolonged the survival of a mouse model of systemic metastasis. These results suggest the possibility of molecular-targeted therapy using cetuximab for endometrial cancer.

The main mode of progression of ovarian cancer is peritoneal dissemination, and its inhibition may lead to improved outcome. Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) reportedly inhibits the proliferation, migration, and invasion of cancer cells. The purpose of this study is to explore the possibility of PTEN gene therapy for ovarian cancer. We transfected the ovarian cancer cell line SHIN-3 [vascular endothelial growth factor (VEGF)-hypersecretory cell line] with PTEN or luciferase (LUC)-expressing plasmid. After selection, PTEN-overexpressing cells (SHIN-3/PTEN) and control cells (SHIN-3/LUC) were obtained. SHIN-3/PTEN implanted s.c. into nude mice was examined for the change in tumor diameter and the number of new blood vessels. Mice with peritoneally disseminated tumors created by i.p. inoculation of the same cells were examined for changes in body weight and abdominal circumference and for survival time. The growth of s.c. SHIN-3/PTEN was significantly lower than that of control (P < 0.001). Compared with controls, mice with i.p. inoculated SHIN-3/PTEN showed significantly smaller increases in the body weight and abdominal circumference (P < 0.01) and a significantly longer survival time (P < 0.05). VEGF concentration in the supernatant of SHIN-3/PTEN was about half that of controls (P < 0.05). The number of new blood vessels in SHIN-3/PTEN was significantly smaller than that in controls (P < 0.001). Overexpression of PTEN suppressed tumor growth and peritoneal dissemination of VEGF-hypersecretory ovarian cancer cells and prolonged the survival time of the mice with peritoneal disseminated tumor. PTEN gene therapy could have therapeutic potential for ovarian cancer and exerts some of this effect by inhibiting angiogenesis.

TSU-68 is a small-molecular-weight synthetic inhibitor of the tyrosine kinase receptors Flk-1/KDR, PDGFRβ and FGFR1, which are involved in angiogenesis. Using a mouse model in which endometrial cancer was subcutaneously implanted, we investigated the effects of TSU-68 alone or in combination with paclitaxel. We subcutaneously implanted a cell strain of endometrial cancer, HEC1A, into BALB/c nude mice. TSU-68 was orally administered every day, while paclitaxel was intraperitoneally injected once a week, and the rates of subcutaneous tumor proliferation were compared. In a group treated with high-dose (200 mg/kg/day) TSU-68 alone, subcutaneous tumor proliferation was significantly inhibited in comparison with a vehicle-treated control group (p<0.05). In groups treated with low-dose TSU-68 or paclitaxel alone (100 and 10 mg/kg/day, respectively), tumor proliferation was not significantly inhibited. In a low-dose combination therapy group (100 mg/kg/day of TSU-68 + 10 mg/kg/day of paclitaxel), tumor proliferation was significantly inhibited in comparison with the control and low-dose TSU-68 or paclitaxel therapy groups (p<0.01). High-dose monotherapy with TSU-68 inhibited the proliferation of the subcutaneously implanted tumor. Furthermore, a combination of TSU-68 and paclitaxel at a low dose, one at which respective monotherapy was not effective, inhibited tumor proliferation. Combination therapy with the two agents may therefore be useful for treating endometrial cancer.

Vascular endothelial growth factor (VEGF) is known to play a major role in angiogenesis in a variety of tumors. A soluble form of Flt-1 (sFlt-1), a VEGF receptor, is potentially useful as an antagonist of VEGF, and accumulating evidences suggest the applicability of sFlt-1 in tumor suppression by means of anti-angiogenesis. We previously demonstrated the efficacy of sflt-1 gene expression in situ to suppress tumor growth and ascites in ovarian cancer. Here, we demonstrate the therapeutic applicability of muscle-mediated expression of sFlt-1 in tumor-bearing mice. Initially, tumor suppressive action was confirmed by inoculating sFlt-1-expressing ovarian cancer (SHIN-3) cells into mice, both subcutaneously and intraperitoneally. To validate the therapeutic efficacy in a more clinically relevant model, adeno-associated virus vectors encoding sflt-1 were introduced into mouse skeletal muscles and were subsequently inoculated with tumor cells. As a result, high serum sFlt-1 levels were constantly observed, and the growth of both subcutaneously- and intraperitoneally-inoculated tumors was significantly suppressed. No delay in wound healing or adverse events of neuromuscular damage were noted, body weight did not change, and laboratory data, such as those representing liver and renal functions, were not affected. These results indicate that sFlt-1 suppresses growth and peritoneal dissemination of ovarian cancer by the inhibition of angiogenesis, and thus suggest the usefulness of gene therapy for ovarian cancer.

SU6668 (TSU-68) is a small-molecule synthetic inhibitor of the angiogenic related receptor tyrosine kinases Flk-1/KDR, PDGFRbeta, and FGFR1. Using a mouse model of peritoneally disseminated ovarian cancer, we investigated whether SU6668 inhibits peritoneal dissemination and prolongs survival time. BALB/c nude mice were intraperitoneally (i.p.) inoculated with SHIN-3 (VEGF-hypersecretory) or KOC-2S (PDGF-hypersecretory) ovarian serous adenocarcinoma cells with marked peritoneal dissemination ability. From the day after i.p. inoculation of tumor cells, SU6668 was orally administered 6 times weekly at a daily dose of 100 mg/kg or 400 mg/kg. The SU6668-administered group and the vehicle-administered control group were compared for the number of tumor vascular endothelial cells, weight of peritoneally disseminated tumors, amount of ascitic fluid and survival time. As a result, these 3 parameters were significantly smaller in the SHIN-3-inoculated, SU6668-administered mice than in the control group (p = 0.03, p = 0.002, and p = 0.02, respectively). The mean survival time was significantly longer, at 58.1 +/- 11.2 days, in the SU6668-administered mice than that (34.5 +/- 8.8 days) in the control group (p = 0.002). Similarly, in the KOC-2S-inoculated mice, the oral administration of SU6668 significantly reduced these 3 parameters (p = 0.04, p = 0.04, and p = 0.03, respectively), and significantly prolonged survival (16.6 +/- 1.7 days vs. 11.0 +/- 0.7 days, p = 0.008). Thus, the oral administration of SU6668 inhibited angiogenesis and peritoneal dissemination and prolonged survival in mice with peritoneally disseminated ovarian cancer. These effects were observed with both the VEGF- and PDGF-hypersecretory cell lines. Our results suggest that molecular targeting with oral SU6668 will become a new therapeutic strategy targeting peritoneally disseminated ovarian cancer.

A Kunitz-type protease inhibitor, bikunin, is known to suppress the invasion and metastasis of cancer cells. HI8, a carboxyl-terminal domain of bikunin, is an active site of this glycoprotein. To increase its affinity for cancer cells, we constructed a chimeric gene, ATF-HI8, and investigated the anti-invasive and anti-migratory activity of ATF-HI8 on ovarian cancer cells. ATF-HI8-expressing plasmid and ATF-expressing plasmid were introduced into the highly invasive and metastatic ovarian cancer cell line HRA. The properties of the established cell line (HRA/ATF-HI8) were compared to those of the HRA/ATF and the HRA/luciferase (HRA/LUC, control) cell lines in terms of cell proliferation, invasion and migration. As a result, (i) there were no differences in cell proliferation between HRA/ATF-HI8 and HRA/LUC; (ii) the invasion and migration of HRA/ATF-HI8 cells were significantly inhibited compared to those of HRA/LUC cells; (iii) the migration, but not the invasion, of HRA/ATF cells was significantly inhibited compared to that of HRA/LUC. These results indicate that the overexpression of ATF-HI8 inhibits the invasion and migration of ovarian cancer cells without affecting cell proliferation and suggest that HI8 is involved in the anti-invasive and the anti-migratory activities, and the addition of ATF brought about the increase in the anti-migratory activity of HI8. The above findings suggest the applicability of therapeutic strategies targeting the inhibition of peritoneal invasion and dissemination of ovarian cancer by the use of the chimeric gene ATF-HI8.

Peritoneal dissemination is the major progression pathway of ovarian cancer, and its control is important for improvement of the prognosis. PTEN is a tumor suppressor gene, and is known to inhibit cancer cell growth and migration. To investigate the possibility of gene therapy using PTEN for ovarian cancer, we introduced PTEN cDNA into an ovarian cancer cell line HRA carrying wild-type PTEN, and examined the effects in vitro and in vivo. Using PTEN cDNA cloned from a human liver cDNA library, a PTEN expression vector was constructed. This vector was introduced into HRA cells by the standard calcium phosphate precipitation method, and an HRA cell line overexpressing PTEN (HRA/PTEN) was established. On the cell migration test by in vitro scratch wound healing assay, the number of migrating cells was 6.3+/-0.9 cells/mm(2) in HRA/PTEN, which was significantly smaller than that in the control (39.7+/-3.2 cells/mm(2)) (p<0.01). No significant differences were observed in the in vitro cell growth or in vivo tumor growth between HRA/PTEN and the control. The findings described above, show that enhanced expression of PTEN inhibits ovarian cancer cell migration, suggesting that gene therapy approaches using PTEN for control of peritoneal dissemination of ovarian cancer are possible.

The prognosis of cancers of various organs overexpressing thymidylate synthase (TS) has been reported to be poor. It has been suggested that the poor prognosis is partly due to a low sensitivity of TS-overexpressing tumors to TS-targeting 5-fluorouracil (5-FU). To investigate the relationship between TS expression and sensitivity to 5-FU, we used the TS-overexpressing cervical cancer cell line SKG-II/TS and SKG-I/TS that had been established by TS gene transfer. The 50% growth inhibitory concentration (IC(50)) of 5-FU for SKG-II/TS was 24 +/- 6.0 microM, which was 6 times as high as that for the control (4.0 +/- 1.1 microM), showing significantly decreased sensitivity to 5-FU (p < 0.01). The IC(50) of 5-FU for SKG-I/TS was 90 +/- 15 microM, which was over 2 times as high as that for the control (40 +/- 0.6 microM), showing significantly decreased sensitivity to 5-FU (p < 0.05). Thus, TS-overexpressing tumors have decreased sensitivity to 5-FU, which may be one of the factors that determine the prognosis of these tumors.

Interleukin-10 (IL-10) is an immunosuppressive cytokine produced by T lymphocytes and drawing attention as an inhibitor of tumor angiogenesis. In this study, we investigated antiangiogenic and tumor suppressive effects of IL-10 in ovarian cancer cells. mIL-10-expressing plasmid was transferred into two ovarian cancer cell lines, SHIN-3 [vascular endothelial growth factor (VEGF) producing] and KOC-2S (non-VEGF producing). After selection, mIL-10-expressing cells were obtained as SHIN-3/mIL-10 and KOC-2S/mIL-10. No significant differences were observed in in vitro growth properties between mIL-10-expressing cells and control (luciferase expressing) cells in either KOC-2S or SHIN-3. The angiogenic activities of mIL-10-expressing cells were measured by dorsal air sac assay, which detected the number of newly formed blood vessels within a chamber in vivo. In addition, tumor formation was evaluated by s.c. tumor transplantation, and survival was monitored after i.p. injection of ovarian cancer cells into BALB/c nude mice. Both in vivo angiogenic activity and tumor growth were significantly inhibited in SHIN-3/mIL-10 cells compared with the control. Moreover, peritoneal dissemination was inhibited, and the survival period was significantly prolonged (mean survival days > 90 versus 36). In contrast, in the case of KOC-2S cells, no significant differences were observed in any of the parameters tested. These results indicate that IL-10 has suppressive effects on angiogenesis, tumor growth, and peritoneal dissemination of VEGF-producing ovarian cancer cells. Although the mechanisms of the antiangiogenic effect of IL-10 are still unclear, the potential usefulness of IL-10-mediated gene therapy of ovarian cancer was suggested.

PROBLEM: Since transvaginal hydrolaparoscopy (THL) was introduced as the first-line procedure in the early stages of the exploration of the adnexal structures in infertile women, it has been shown that THL is a less traumatic and a more suitable outpatient procedure than diagnostic laparoscopy. This study was performed to investigate the relationships between Chlamydia trachomatis antibody titers and tubal pathology assessed using THL in infertile women. METHODS: The C. trachomatis antibody titers (IgG and IgA) were evaluated by ELISA. The posterior of the uterus and the tubo-ovarian structures were carefully observed, and tubal passage using indigocarmine was confirmed using THL. THL was carried out in 32 infertile women having C. trachomatis antibody in their sera between May 1999 and October 2001. Unilateral salpingectomy had been performed on two of the 32 patients. RESULTS: Tubal occlusion was confirmed in 20 (32.3%) of the 62 tubes, while peritubal adhesion was diagnosed in 37 (59.7%) of the 62 tubes. Using receiver operating characteristics curves, the cut-off value of C. trachomatis IgG antibody titer to predict tubal occlusion was determined to be 3.55. Tubal occlusion was observed in 16 (51.6%) of the 31 tubes in patients with the C. trachomatis IgG antibody titer of more than 3.55, which was significantly higher in four (12.9%) of the 31 tubes having the antibody titer less than 3.55 (P = 0.004). However, there was no correlation between C. trachomatis IgG antibody titer and peritubal adhesion. As for C. trachomatis IgA antibody titer, there was no correlation between antibody titer and tubal occlusion or peritubal adhesion. CONCLUSIONS: These results suggest that C. trachomatis infection is significantly associated with tubal pathology. Although the cut-off value of C. trachomatis IgG antibody titer to predict the existence of tubal occlusion was shown to be 3.55, we would suggest that THL or standard laparoscopy is performed to consider appropriate treatments in patients with past C. trachomatis infection because of the high prevalence of peritubal adhesion.

Bikunin, a Kunitz-type protease inhibitor, could potentially suppress tumor cell invasion and metastasis. Our previous study revealed that overexpression of bikunin in a human ovarian cancer cell line, HRA, resulted in a down-regulation in uPA and uPAR gene expression. For identifying the full repertoire of bikunin-regulated genes, a cDNA microarray hybridization screening was conducted using mRNA from bikunin-treated or bikunin-transfected HRA cells. A number of bikunin-regulated genes were identified, and their regulation was confirmed by Northern blot analysis. Our screen identified 11 bikunin-stimulated genes and 29 bikunin-repressed genes. The identified genes can indeed be classified into distinct subsets. These include transcriptional regulators, oncogenes/tumor suppressor genes, signaling molecules, growth/cell cycle, invasion/metastasis, cytokines, apoptosis, ion channels, extracellular matrix proteins, as well as some proteases. This screen identified suppression of several genes such as CDC-like kinase, LIM domain binding, Ets domain transcription factor, Rho GTPase-activating protein, tyrosine phosphorylation-regulated kinase, hyaluronan-binding protein, matriptase, and pregnancy-associated plasma protein-A (PAPP-A), which have previously been implicated in enhancing tumor promotion. Northern blot analysis confirmed that several genes including matriptase and PAPP-A were down-regulated by bikunin by approximately 9-fold. Further, genetic inhibition of matriptase or PAPP-A could lead to diminished invasion. These results show that bikunin alters the pattern of gene expression in HRA cells leading to a block in cell invasion.

Bikunin (bik), a Kunitz-type protease inhibitor, also known as urinary trypsin inhibitor, is proposed as a main participant in the inhibition of tumor cell invasion and metastasis, possibly through the direct inhibition of cell-associated plasmin activity and suppression of urokinase-type plasminogen activator (uPA) mRNA expression. In the present study, we transfected the human ovarian carcinoma cell line HRA, highly invasive cells, with an expression vector harboring a cDNA encoding for human bik. Our study was designed to investigate the effect of bik overexpression and changes in tumor cell phenotype and invasiveness in the stably transfected clones. Bik gene transfection of HRA gave the following results: 1) transfection of HRA with the bik cDNA resulted in 5 variants stably expressing functional bik; 2) bik(+) clones exhibited a significantly reduced uPA mRNA expression as compared to the parental cells; 3) bikunin negatively regulates the ERK1/2 activity; 4) secretion-blocking treatments of bik(+) clones abrogated bik-mediated suppression of ERK1/2 activation and uPA expression; 5) the regulation of invasion seen in the HRA cells is mainly mediated by the uPA-plasmin-MMP-2 system; 6) transfection of HRA with the bik gene significantly reduced invasion, but not proliferation, adhesion, or migration relative to the parental cells; and 7) animals with bik(+) clones induced reduced peritoneal dissemination and long term survival. We conclude that transfection of HRA cells with the bik cDNA constitutively suppresses ERK1/2 activation, which results in inhibition of uPA expression and subsequently reduces dissemination of bik(+) clones.