Metronomic irinotecan chemotherapy combined with ultrasound irradiation for a human uterine sarcoma xenograft.

Metronomic chemotherapy is the frequent administration of low doses of chemotherapeutic agents targeting tumor-associated endothelial cells. We examined the efficacy of metronomic irinotecan combined with low-intensity ultrasound (US) in human uterine sarcoma and evaluated its antiangiogenesis mechanism by measuring the circulating endothelial progenitor cells (CEP), a surrogate marker of angiogenesis. A human uterine sarcoma cell line, FU-MMT-3, was used in the present study because this tumor is one of the most malignant neoplasms of human solid tumors and it also has a high angiogenesis property. The combination of low-dose irinotecan and US irradiation significantly inhibited the tube formation of HUVEC and vascular endothelial growth factor expression of tumor cells in vitro. The FU-MMT-3 xenografts in nude mice were treated using US at a low intensity (2.0 w/cm(2), 1 MHz) for 4 min three times per week each after the intraperitoneal administration of irinotecan; this treatment was continued for 5 weeks. The tumor vascularity was assessed by contrast-enhanced color Doppler US in real time. The combination treatment significantly inhibited the mobilization of CEP and intratumoral vascularity compared with the control. This combination therapy showed a significant reduction in tumor volume, resulting in a significant prolongation of survival, in comparison with each treatment alone. These results suggest that the effect of metronomic chemotherapy for human uterine sarcoma was accelerated by US irradiation in vivo and this combination might therefore be potentially effective for new cancer therapy.

Metronomic doxifluridine chemotherapy combined with the anti-angiogenic agent TNP-470 inhibits the growth of human uterine carcinosarcoma xenografts.

Uterine carcinosarcoma is a highly aggressive gynecological neoplasm that responds poorly to conventional chemotherapy and radiotherapy. Metronomic chemotherapy is accepted as a new approach for cancer treatment, and its underlying mechanism seems to involve the suppression of angiogenesis. However, the efficacy of metronomic and anti-angiogenic therapies against uterine carcinosarcoma is unknown. The anti-angiogenic effect of doxifluridine was assessed in vitro using human umbilical vein endothelial cells (HUVEC) co-cultured with FU-MMT-1 human uterine carcinosarcoma cells. The antitumor and anti-angiogenic effects of metronomic doxifluridine (delivered via oral gavage) in combination with TNP-470 were evaluated in vivo. Tumor vascularity was assessed by contrast-enhanced color Doppler ultrasound, laser Doppler and microvessel density staining. Doxifluridine suppressed tube formation of HUVEC and vascular endothelial growth factor production by FU-MMT-1 cells. Metronomic doxifluridine alone significantly suppressed tumor growth compared with the untreated (control) group, while metronomic doxifluridine in combination with TNP-470 significantly inhibited tumor growth compared with each treatment alone. A significant reduction of intratumoral vascularity was observed in FU-MMT-1 xenografts following treatment with metronomic doxifluridine in combination with TNP-470, as compared with each treatment alone. Intestinal bleeding was only observed when the maximum tolerated dose of doxifluridine was administered in combination with TNP-470. Metronomic doxifluridine chemotherapy in combination with TNP-470 might be effective for uterine carcinosarcoma without marked toxicity, possibly acting via its potent anti-angiogenic effects. Clinical studies are needed to evaluate the safety and efficacy of this treatment in humans.

Clinical usefulness of contrast-enhanced color Doppler ultrasonography in invasive and noninvasive gestational trophoblastic diseases: a preliminary study.

OBJECTIVE: To examine the usefulness of contrast-enhanced color Doppler ultrasonography (CDU) in differentiating between invasive and noninvasive gestational trophoblastic disease (GTD). STUDY DESIGN: In 23 patients with findings suggestive of GTD by transvaginal gray-scale ultrasonography, the presence or absence of blood flow within uterine lesions was assessed by contrast-enhanced CDU using Levovist (Schering, Berlin, Germany) microbubble contrast agent. Intratumoral blood flow waveforms were analyzed using resistance indices. Tumor size in each invasive or malignant GTD was assessed by magnetic resonance imaging. RESULTS: Intratumoral blood flow was detected in all invasive or malignant GTDs (7/7: 5 invasive moles, 1 choriocarcinoma and 1 placental site trophoblastic tumor), whereas it was not seen in any noninvasive GTD (0/16:10 complete moles, 5 partial moles and 1 exaggerated placental site) (p <0.0001). A marked increase in uterine vascularity was thus shown in all invasive or malignant GTDs following enhancement. In small invasive moles (<2 cm) in the uterine myometrium, color flow was remarkably increased by contrast-enhanced CDU. Intratumoral blood flow waveforms showed low resistance indices in all invasive and malignant GTDs. CONCLUSION: Contrast-enhanced CDU may be useful in differentiating invasive or malignant GTDs from noninvasive GTDs. By enhancing color flow, this minimally invasive approach may be helpful for detecting small invasive GTD lesions within the uterine myometrium.

Novel chemoembolization using calcium-phosphate ceramic microsphere incorporating TNP-470, an anti-angiogenic agent.

The purpose of the present study was to develop a new method of chemoembolization to improve the therapeutic effectiveness and safety profile of cancer treatment. A chemoembolization approach was designed for human solid tumors using resorbable calcium-phosphate ceramic microspheres loaded with an agent anti-angiogenic to tumor vasculature in vivo. The human uterine sarcoma cell line FU-MMT-3 was used in this study because this tumor is aggressive and also exhibits a poor response to radiotherapy or any chemotherapy currently used. The calcium-phosphate ceramic microspheres loaded with TNP-470, an anti-angiogenic agent, were injected into FU-MMT-3 xenografts in nude mice three times per week for 8 weeks. The treatment using TNP-470-loaded microspheres suppressed tumor growth, compared to treatment with TNP-470 alone, microspheres alone, and the control. The mean tumor weight after treatment using TNP-470-loaded microspheres was significantly lower than that after treatment with microspheres alone. These ceramic microspheres were remarkably embolized in tumor microvessels as well as in the feeding arteries and a significant reduction of intratumoral vascularity was also demonstrated following treatment with TNP-470-loaded microspheres. Severe loss of body weight was not observed in any mice treated with the TNP-470-loaded microspheres, compared to treatment with TNP-470 alone. These results suggest that targeting tumor vasculature in human uterine sarcoma using calcium-phosphate microspheres might be more effective and safer than the treatment that employs anti-angiogenic agent alone. This new chemoembolization method incorporating an anti-angiogenic agent may contribute to the effective treatment of locally advanced or recurrent solid tumors.

Primary leiomyosarcoma of the fallopian tube.

Primary sarcoma of the fallopian tube is a very rare neoplasm. We report the case of a 69-year-old woman affected with leiomyosarcoma of the left fallopian tube. Her chief complaint was lower abdominal pain. The preoperative diagnosis was a left adnexal malignant tumor based on pelvic examination, abdominal computed tomography, and magnetic resonance imaging. Following a laparotomy, she was ultimately diagnosed with a FIGO IIc fallopian tube leiomyosarcoma. She was treated with total abdominal hysterectomy, bilateral salpingo-oophorectomy, pelvic lymph node dissection, partial omentectomy, and low anterior resection for rectal invasion. The patient subsequently received adjuvant chemotherapy with pirarubicin and ifosfamide. Thirty months after the first therapy, a computed tomography scan revealed metastasis of the liver, lung, and supraclavicular lymph node. The patient died of the disease 39 months after the initial treatment.

Longitudinal changes in canal length at 16-35 weeks in normal twin pregnancies and twin pregnancies with preterm labor and delivery.

AIM: To describe the longitudinal changes in canal length at 16-35 weeks' gestation in cases of twin pregnancy with preterm labor and delivery. METHODS: We studied 22 cases of twin pregnancy that were delivered at < 36 weeks and/or that underwent preterm labor requiring tocolysis. We also studied 44 cases of twin pregnancy delivered at > or = 36 weeks without tocolysis (non-tocolysis twin pregnancy). Controls were 82 cases of normal singleton pregnancy. Canal length was longitudinally measured using transvaginal ultrasonography. The observational period of 16-35 weeks was divided into 4-week periods for analysis. RESULTS: From 28 to 31 weeks onwards the canal length of non-tocolysis twin pregnancies was shorter than that of normal singleton pregnancies (P < 0.05). The canal length of twin pregnancies with preterm labor and delivery was shorter than that of non-tocolysis twin pregnancies at 16-19 weeks and decreased rapidly until 24-27 weeks (P < 0.01). CONCLUSIONS: A short canal length at 16-19 weeks followed by rapid canal length shortening in the second trimester are specific characteristics in preterm labor and delivery of twin pregnancies. Sequential measurements of canal length in the second trimester starting at < 20 weeks may be a suitable parameter to predict preterm labor and delivery in twin pregnancies.

Monitoring of endometrial K-ras mutation in tamoxifen-treated patients with breast cancer.

INTRODUCTION: A high incidence of endometrial K-ras mutations has been reported in tamoxifen (TAM)-treated patients with breast cancer. We examined the changes in the frequency of the endometrial K-ras mutations after the cessation of TAM treatment. METHODS: DNA was extracted from fresh cytological or polypectomy samples of the endometrium in 28 patients who had undergone TAM treatment of breast cancer. Mutations were detected by an enriched polymerase chain reaction-enzyme-linked minisequence assay (Sumitomo Metal Industry, Inc, Tokyo, Japan). K-ras codon 12 mutations were monitored in these 28 patients. RESULTS: An initial examination detected endometrial K-ras mutations in 13 of the 28 patients. However, repeated examinations performed after cessation of TAM treatment did not detect endometrial K-ras mutations in any of these 13 patients. No endometrial K-ras mutation has been detected in the repeated examinations performed for these patients for more than 2 years since the cessation of TAM treatment. In addition, the 15 patients who did not have endometrial K-ras mutations in the initial examination did not demonstrate them in repeat examinations. CONCLUSIONS: The cessation of TAM treatment may reduce the risk of developing endometrial cancers through K-ras mutations.

Endometrial cytologic findings in tamoxifen-treated breast cancer patients.

OBJECTIVE: To describe the endometrial cytologic findings in tamoxifen (TAM)-treated patients and evaluate the clinical significance of these findings. STUDY DESIGN: A total of 1,739 endometrial cytologic samples were obtained from 203 breast cancer patients with TAM treatment using an endocyte device. Histologic examinations were recommended for the 23 endometrial cytologic samples (18 initial and 5 follow-up endometrial cytologic samples). These 23 samples were cytopathologically reviewed. RESULTS: A large nuclear dimension and anisokaryosis of the endometrial glandular cells were found in the 23 endometrial cytologic samples. All patients were amenorrheic and postmenopausal except for 3 premenopausal women with endometrial cancer. The subsequent histologic examinations showed 3 atrophic endometria, 3 cystic atrophies, 11 endometrial polyps and 6 endometrial cancers. The tumor diathesis and atypical features of cell aggregates, such as a loss of polarity and severe piling up of nuclei, were seen in the cytologic samples of the endometrial cancers but were not found in those with benign conditions. CONCLUSION: A large nuclear dimension and anisokaryosis of the glandular cells were sometimes found in endometrial cytologic samples from the TAM-treated patients. The atypical characteristics of the cell aggregates with these types of cellular atypia are useful for the cytologic assessment of malignancy.

Validation of HB-EGF and amphiregulin as targets for human cancer therapy.

Aberrant expression levels of epidermal growth factor receptor (EGFR) and its cognate ligands have been recognized as one of the causes of cancer progression. To investigate the validity of EGFR ligands as targets for cancer therapy, we examined the expression of EGFR ligands and in vitro anti-tumor effects of small interference RNA (siRNA) for EGFR ligands in various cancer cells. HB-EGF expression was dominantly elevated in ovarian, gastric, and breast cancer, melanoma and glioblastoma cells, whereas amphiregulin was primarily expressed in pancreatic, colon, and prostate cancer, renal cell carcinoma and cholangiocarcinoma cells. Transfection of siRNAs for HB-EGF or amphiregulin into these cells significantly increased the numbers of apoptotic cells with attenuation of EGFR and ERK activation. In lung cancer cells, any EGFR ligand was not recognized as a validated target for cancer therapy. These results suggest that HB-EGF and amphiregulin are promising targets for cancer therapy.

Changes in lipid metabolism in postmenopausal Japanese women using hormone therapy evaluated by capillary isotachophoresis: a pilot study.

OBJECTIVE: The aim of this study was to clarify the changes in lipid metabolism in postmenopausal Japanese women during continuous combined hormone therapy (HT) in detail by using capillary isotachophoresis (cITP). DESIGN: Twenty-three postmenopausal Japanese women with climacteric symptoms were recruited. Blood samples were collected from all participants before HT and after 3 and 6 months of HT, and changes in total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, and apolipoprotein (apo) A-I, apoB, and lipoprotein(a) were assessed. The same blood samples were analyzed for charge-based lipoprotein subfractions using cITP. RESULTS: Total cholesterol (-8.5%), LDL-cholesterol (-16.2%), and lipoprotein(a) (-25.5%) decreased and apoA-I (9.5%) increased significantly from the baseline level at 6 months after starting continuous combined HT. HDL-cholesterol increased nonsignificantly (+2.4%). cITP revealed that the fast HDL subfraction (+24.7%), which contains apoA-I and may be antiatherogenic, significantly increased. Conversely, the fast LDL subfraction (+14.4%), which may possess atherogenic potential, increased during HT. CONCLUSIONS: The changes in lipid metabolism in postmenopausal Japanese women receiving HT are inconclusive regarding atherogenicity, ie, it increased fast HDL and simultaneously increased fast LDL. cITP is useful for clarifying the changes in lipid metabolism during HT.

Modification of the endoscopic management of congenital duodenal stenosis.

This report documents a new endoscopic management modality for congenital membranous stenosis in the third portion of the duodenum. Standard approaches to duodenal stenosis in newborns include a laparotomy with an enteroenterostomy, bypassing the obstruction, or a duodenoduodenostomy with excision. We successfully developed a modification of the endoscopic treatment modality for congenital duodenal diaphragm.

[The efficacy and safety of indisetron hydrochloride for the management of nausea/vomiting caused by chemotherapy for gynecologic cancer].

This clinical trial was designed to evaluate the efficacy and safety of indisetron hydrochloride an oral 5-HT3 receptor antagonist, for the management of nausea/vomiting caused by chemotherapy for gynecologic cancer with paclitaxel/ carboplatin or docetaxel/carboplatin. Indisetron hydrochloride(8 mg)was administered orally to 46 gynecologic cancer patients at 0.5 hours before administration of the above chemotherapy agents. Number of patients who showed nausea or vomiting for 24 hours was counted. The complete vomiting inhibition rate at 24 hours after chemotherapy was 89.1%(41/46), and nausea inhibition rate was 71.7%(33/46). No serious adverse events were observed. These findings indicate that prophylactic administration of indisetron hydrochloride is safe and useful for inhibition of nausea/vomiting caused by cancer chemotherapy.

New approach to cancer therapy: heparin binding-epidermal growth factor-like growth factor as a novel targeting molecule.

Heparin binding-epidermal growth factor-like growth factor (HB-EGF) is one of the EGF receptor ligands and possesses several functional domains. It is involved in diverse biological processes, including wound healing, blast implantation, atherosclerosis and tumor formation, through its interactions with various molecules. We have reported that HB-EGF gene expression is significantly elevated in human ovarian cancer, and further demonstrated that HB-EGF plays key roles in the acquisition of malignant phenotypes, such as cell survival in peritoneal fluid, cell adhesion on extracellular matrices, invasion, angiogenesis, tumorigenicity, and chemoresistance in ovarian cancer. Thus, HB-EGF was considered as a promising target for cancer therapy. In vitro as well as in vivo experiments have revealed that cross-reacting material 197 (CRMI97), a specific inhibitor of HB-EGF, or a small interfering RNA for HB-EGF can block each step involved in peritoneal dissemination. According to these pieces of evidence, the development of targeting tools against HB-EGF, such as CRM197, could allow us to improve the prognosis of cancer patients.

Overexpression and diffuse expression pattern of IQGAP1 at invasion fronts are independent prognostic parameters in ovarian carcinomas.

IQGAP1 is a multifunctional protein involved in actin cytoskeleton assembly and E-cadherin-mediated cell adhesion. To determine the role of IQGAP1 in ovarian tumors, we evaluated IQGAP1 expression by immunohistochemistry in 17 adenomas, 30 borderline tumors and 80 adenocarcinomas and its relation with patient survival. IQGAP1 was overexpressed in adenocarcinomas compared with adenomas and borderline tumors. Enhanced immunostaining in invasive tumor fronts was categorized as focal or diffuse. The diffuse expression pattern correlated with high histological grade and clinicopathological stages. IQGAP1 overexpression and diffuse invasion pattern were significantly associated with poor prognosis by multivariate analysis. Our findings suggest the involvement of IQGAP1 in the progression and spread of ovarian adenocarcinomas. Overexpression and diffuse expression pattern of IQGAP1 are potentially useful independent molecular predictors of highly aggressive ovarian carcinomas.

Sonographic findings of transvaginal color Doppler ultrasound in ectopic pregnancy.

PURPOSE: The objective of this study was to retrospectively assess whether the sonographic findings from transvaginal color Doppler ultrasound (TV-CDU) are helpful in the diagnosis of ectopic pregnancy. METHODS: Thirty-four patients who received surgery for ectopic tubal pregnancies were preoperatively evaluated using TV-CDU. The presence or absence of color vascularity within the ectopic masses was examined. The relationship between the presence or absence of blood flow in the tubal mass and the corpus luteum cyst, or the serum ß-hCG values, was evaluated. RESULTS: Color vascularity within the adnexal mass was detected in 27 of 34 (79.4%) patients with ectopic pregnancies by TV-CDU. Color vascularity within the mass was observed in 18 of 24 (75.0%) patients with a questionable adnexal mass that had no obvious gestational sac in B-mode images. Moreover, color vascularity was seen in all four patients with a serum ß-human chorionic gonadotropin (ß-hCG) value of less than 500 mIU/ml. However, it was difficult to identify the blood flow of the adnexal mass in six of the nine (66.7%) patients with a corpus luteum cyst in the ipsilateral ovary. No relationship was observed between the serum ß-hCG concentrations and the resistance indices, or the peak systolic velocity. CONCLUSIONS: The detection of color vascularity by TV-CDU in patients with an ectopic pregnancy is helpful for diagnosis, especially for patients with either a questionable adnexal mass in B-mode images or lower serum ß-hCG concentrations.

Early human preantral follicles have relaxin and relaxin receptor (LGR7), and relaxin promotes their development.

The regulatory mechanisms of early follicle development are not clearly understood. Although relaxin is a peptide that controls cell proliferation and differentiation in many tissues, its role in human follicular development is unclear. In this study we cultured slices of human ovarian cortical tissue in the presence and absence of recombinant human relaxin. Ovarian tissue was obtained by biopsy during gynecological laparotomy or laparoscopy (14 women; mean age +/- sem, 29.0 +/- 6.1 yr; range, 17-37 yr). A significantly higher proportion of secondary follicles (14.5% vs. 5.0% in the control group; P < 0.01) and a significantly decreased proportion of primordial follicles (30.1% vs. 47.4% in the control group; P < 0.05) were found in tissues cultured with relaxin for 7 d. Immunocytochemical studies with the anti-C-peptide of prorelaxin and antirelaxin antibodies revealed the localization of relaxin in the oocyte and in flat pregranulosa and granulosa cells of primordial, primary, and secondary follicles. The presence of the relaxin receptor LGR7 was observed in flat pregranulosa and granulosa cells of primordial, primary, and secondary follicles by immunocytochemical and in situ hybridization analyses. These results suggest that relaxin plays a role through its receptor during the early stage of follicle development.

The combination of Jun N-terminal kinase inhibitor and TNP-470 blocks carcinosarcoma-induced endothelial cell tube formation in a synergistic manner.

We assessed the usefulness of Jun N-terminal kinase inhibitor (JNK-I) as an anti-angiogenic agent against a human uterine carcinosarcoma cell line (FU-MMT-1). JNK-I blocked FU-MMT-1-induced human arterial endothelial cell (HAEC) tube formation in an in vitro co-culture model. Cell proliferation of FU-MMT-1 or HAEC was inhibited by JNK-I. In addition, JNK-I blocked matrix metalloproteinase production but not vascular endothelial growth factor (VEGF) secretion in HAECs. Although low concentrations of JNK-I or TNP-470, an anti-cancer agent, did not separately block FU-MMT-1-induced tube formation, such tube formation was blocked by the combination of low concentrations of JNK-I and TNP-470 because TNP-470 blocked VEGF production, suggesting that JNK-I and TNP-470 had a synergistic effect and might be effective in patients with carcinosarcoma.

Synchronous endometrial and ovarian carcinomas: analysis of genetic relationship of the tumors.

Simultaneous carcinomas of the endometrium and ovary may represent a diagnostic dilemma and the clinical management of such cases may be problematic. In surgical pathology practice, we classify them either as double primary tumors (DP) or a single primary tumor with metastasis (PM), according to the conventional clinicopathologic criteria. The distinction has important therapeutic and prognostic implications, but it can be sometimes difficult, especially in advanced cases. In this study, in addition to the clinicopatho-logic classification, we assessed tumor cell clonality in 13 cases with synchronous endometrial and ovarian endometrioid adenocarcinomas using PCR-based microsatellite analysis of microdissected archival tissues for loss of heterozygosity (LOH) and microsatellite instability (MSI). All paired endometrial and ovarian tumors demonstrated either MSI or/and LOH except for 1 case, and therefore the microsatellite analysis was informative in 92.3% of the cases. Nine of 26 tumors (34.6%) exhibited MSI-H and 15 of 26 (57.7%) showed LOH. In contrast to 4 DP cases and 9 PM cases classified according to clinicopathologic criteria, microsatellite analysis suggested 10 DP cases and 2 PM cases. The molecular analysis was not informative in 1 case. Thus, analysis of microsatellite abnormality is a helpful adjunct in the assessment of synchronous tumors, especially to differentiate DP from PM cases in advanced tumor cases. Moreover, the combination of conventional clinicopathologic evaluation and molecular analysis is important and helpful in distinction between the two groups of synchronous tumors.

Relaxin-induced angiogenesis in ovary contributes to follicle development.

CD31-positive blood vessels increased in human ovarian cortical tissues cultured with relaxin than in tissues cultured without relaxin. The number of vWF-positive vessels counted in rat ovaries after relaxin injection (40.5 +/- 4.3) significantly increased compared to that in saline-injected rats (24.3 +/- 3.0). The proportion of primordial follicles was significantly decreased, that of primary follicles was significantly increased at 16 h, and that of early antral follicles was significantly increased at 24 h in rats after relaxin injection. These results suggest that relaxin may contribute to follicle development through the mechanism of angiogenesis in the ovary.