Association of Serum HB-EGF Value and Response to Chemotherapy in Patients with Recurrent Ovarian Cancer.

BACKGROUND/AIM: Many anticancer agents including molecularly-targeted drugs have been developed for ovarian cancer. However, the prognosis of recurrent ovarian cancer remains extremely poor. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is reported as a rational target for ovarian cancer therapy. Moreover, serum HB-EGF expression is recognized as a biomarker in patients with primary ovarian cancer. MATERIALS AND METHODS: We analysed serum samples with recurrent ovarian cancer at the Fukuoka University Hospital from April 2009 to March 2014. To assess the clinical significance of serum HB-EGF in recurrent ovarian cancer, the association between serum HB-EGF levels and prognosis in patients with recurrent ovarian cancer was examined using ELISA. RESULTS: Patients with high serum HB-EGF expression showed a significantly poor response to second-line chemotherapeutic agents compared with patients with low HB-EGF levels. CONCLUSION: HB-EGF expression in serum may be a potential therapeutic indicator for novel HB-EGF-targeted therapy in recurrent ovarian cancer.

Serum Heparin-binding Epidermal Growth Factor-like Growth Factor (HB-EGF) as a Biomarker for Primary Ovarian Cancer.

Ovarian cancer is the most lethal malignancy among gynaecological cancers. Although many anticancer agents have been developed for the treatment of ovarian cancer, it continues to have an extremely poor prognosis. Heparin-binding epidermal growth factor-like grown factor (HB-EGF) has been reported to be a rational therapeutic target for ovarian cancer. Here, we evaluated the clinical significance of serum HB-EGF by examining the association between prognosis and serum HB-EGF levels in patients with primary ovarian cancer. We found that high serum HB-EGF concentrations were significantly associated with poor prognosis in a combined cohort of patients with all stages of ovarian cancer, as well as in a subset of patients with advanced disease. In addition, serum HB-EGF levels increased as the cancer advanced. These data suggest that serum HB-EGF may be a target for the design of novel therapies for ovarian cancer.

BK-UM in patients with recurrent ovarian cancer or peritoneal cancer: a first-in-human phase-I study.

BACKGROUND: BK-UM (CRM197) is a mutant form of diphtheria toxin and a specific inhibitor of heparin-binding epidermal growth factor-like growth factor (HB-EGF). We assessed the safety, pharmacokinetics, recommended dose, and efficacy of BK-UM in patients with recurrent ovarian cancer (OC) or peritoneal cancer (PC), and measured HB-EGF levels in serum and abdominal fluid after BK-UM administration. METHODS: Eleven patients with advanced or recurrent OC or PC were enrolled and treated with BK-UM via the intraperitoneal route. The dose was escalated (1.0, 2.0, 3.3, and 5.0 mg/m2) using a 3 + 3 design. RESULTS: Eight of 11 patients completed treatment. No dose-limiting toxicity (DLT) was experienced at dose levels 1 (1.0 mg/m2) and 2 (2.0 mg/m2). Grade 3 transient hypotension as an adverse event (defined as a DLT in the present study) was observed in two of four patients at dose level 3 (3.3 mg/m2). Treatment with BK-UM was associated with decreases in HB-EGF levels in serum and abdominal fluid in seven of 11 patients and five of eight patients, respectively. Clinical outcomes included a partial response in one patient, stable disease in five patients, and progressive disease in five patients. CONCLUSIONS: BK-UM was well tolerated at doses of 1.0 and 2.0 mg/m2, with evidence for clinical efficacy in patients with recurrent OC or PC. A dose of 2.0 mg/m2 BK-UM is recommended for subsequent clinical trials. TRIAL REGISTRATION: This trial was prospectively performed as an investigator-initiated clinical trial. The trial numbers are UMIN000001002 and UMIN000001001, with registration dates of 1/30/2008 and 2/4/2008, respectively. UMIN000001001 was registered as a trial for the continuous administration of BK-UM after UMIN000001002 .

MicroRNA-135a-3p as a promising biomarker and nucleic acid therapeutic agent for ovarian cancer.

Ovarian cancer is the most lethal gynecologic malignancy. Recently, several molecularly targeted anticancer agents have been developed for ovarian cancer; however, its prognosis remains extremely poor. The development of molecularly targeted therapy, as well as companion diagnostics, is required to improve outcomes for patients with ovarian cancer. In this study, to identify microRNAs (miRNAs) involved in the progression of ovarian cancer we analyzed serum miRNAs in patients with ovarian cancer using miRNA array and quantitative RT-PCR and examined the anticancer properties of miRNA expression in ovarian cancer cells. In patients with ovarian cancer, high amount of miR-135a-3p in serum samples was significantly associated with favorable clinical prognosis. The amount of miR-135a-3p was significantly decreased in patients with ovarian cancer compared with patients with ovarian cysts or normal ovaries. In SKOV-3 and ES-2 human ovarian cancer cells, enhanced expression of miR-135a-3p induced drug sensitivity to cisplatin and paclitaxel and suppressed cell proliferation and xenograft tumor growth. These findings suggest that miR-135a-3p may be considered as a biomarker and a therapeutic agent in ovarian cancer.

Separation efficiency of a microfluidic sperm sorter to minimize sperm DNA damage.

OBJECTIVE: To evaluate whether microfluidic sperm sorters (MFSSs) allow effective recovery of sorted motile sperm without DNA damage compared with the centrifugation and swim-up procedure. DESIGN: Experimental laboratory study. All participants completed questionnaires regarding previous and/or current diseases, surgery, reproductive experiences, lifestyle factors, and date of the preceding ejaculation. SETTING: University research laboratory. PATIENT(S): Male volunteers were recruited without setting conditions. Semen samples from healthy volunteers (n = 37) were collected in sterile containers by masturbation. INTERVENTION(S): Flow cytometric measurement and sperm chromatin structure assay analysis of DNA damage after sperm preparation using MFSS and the centrifugation and swim-up procedure. MAIN OUTCOME MEASURE(S): Efficacy and efficiency of sperm preparation, correlation between sperm DNA fragmentation index (DFI) and semen parameters, and relationship between basic characteristics and DFI after the centrifugation and swim-up procedure. RESULT(S): Final sperm concentration and motility were significantly different between the centrifugation and swim-up procedure and MFSS sperm preparations. A significantly lower sperm DNA fragmentation rate was detected with MFSS compared with the centrifugation and swim-up procedure use. No correlation was observed between DFI and smoking or drinking, but significant correlations were observed between DFI and medication use and sexual abstinence duration. CONCLUSION(S): MFSSs can be used to efficiently and reliably prepare sperm compared with the centrifugation and swim-up procedure. Further research on the clinical use of MFSSs is required to evaluate the safety and usefulness of this device.

Proposal of a new parameter for diagnosing a low-lying placenta in the third trimester: The distance between the external os and placental edge.

OBJECTIVE: To validate a new parameter of the distance between the external os (EO) and placental edge (PE) to diagnose a low-lying placenta in the third trimester. MATERIALS AND METHODS: The study participants included 94 uncomplicated singleton pregnant women with cephalic presentation. These women were cared for in our hospital in 1998-2011, with a posterior low-lying placenta, which was diagnosed as the distance between the internal os (IO) and a PE of 0-3.0 cm at 34-36 weeks' gestation. Measurements of cervical length (CL) and the distances of IO-PE and EO-PE were performed using transvaginal ultrasonography at least twice at 28-30 weeks, 31-33 weeks, and 34-36 weeks. Changes in CL, and the IO-PE and EO-PE distances were analyzed. RESULTS: CL and the IO-PE and EO-PE distances did not change prior to 31-33 weeks. CL was shortened and the IO-PE distance was increased after 31-33 weeks (p = 0.0001), but the EO-PE distance was unchanged. CONCLUSION: The EO-PE distance is a promising parameter for diagnosis of low-lying placenta in the third trimester up to 36 weeks' gestation.

Clinical significance of cell-free and concentrated ascites re-infusion therapy for advanced and recurrent gynecological cancer.

BACKGROUND: The management of malignant ascites is critical for the treatment of patients with advanced gynecological cancer. The purpose of this study was to assess the clinical significance of cell-free and concentrated ascites re-infusion therapy (CART). PATIENTS AND METHODS: Adverse events, alterations in Eastern Cooperative Oncology Group performance status, serum albumin, body weight and abdominal circumference, and overall survival were examined in 22 patients with advanced gynecological cancer which were treated with CART. RESULTS: Most of the adverse events were grade 1 or 2 fever. CART treatment had little effect on ECOG performance status and on levels of serum albumin. There was a significant decrease in body weight and in abdominal circumference post-treatment with CART, relative to pre-treatment (p<0.01). The overall survival rate was significantly prolonged in 14 patients after CART plus chemotherapy, as compared with eight patients after CART alone (p<0.01). CONCLUSION: CART may contribute to the improvement of quality of life and of survival in patients with advanced gynecological cancer.

Assessment of HB-EGF levels in peritoneal fluid and serum of ovarian cancer patients using ELISA.

BACKGROUND: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a rational target for ovarian cancer therapy. The aim of this study was to examine HB-EGF levels in the peritoneal fluid and serum of ovarian cancer (OVCA) patients. PATIENTS AND METHODS: Samples were collected from six healthy women, 21 OVCA patients, and 21 ovarian cyst patients. HB-EGF levels were measured using a sandwich ELISA kit and calculated using a parallel line assay. RESULTS: No significant difference between the slopes of the standard and sample curves was observed at an anti-HB-EGF antibody concentration of 1.6 µg/ml. HB-EGF levels in the peritoneal fluid and serum of OVCA patients were significantly higher than those in patients with ovarian cysts or controls. Serum HB-EGF levels were also significantly correlated with levels in peritoneal fluid in OVCA patients. CONCLUSION: We developed an assay for the exact measurement of HB-EGF levels in peritoneal fluid and serum.

Uterine inversion due to a leiomyoma on postpartum day 41: a case report.

Uterine leiomyomas are common tumors in women of reproductive age and are frequently detected during pregnancy. The major complications during pregnancy include abortion, preterm delivery, abruptio placentae, intrauterine growth retardation, dystocia, and postpartum hemorrhage. Little attention is given to uterine leiomyomas postpartum compared to leiomyomas prior to childbirth. In the present case, a 27-year-old woman, gravida 1 para 1, presented with massive vaginal bleeding, urinary retention and lower abdominal pain on postpartum day 41. She was diagnosed with uterine inversion due to leiomyoma. After a vaginal myomectomy, the uterus was re-placed with a combined vaginal and abdominal approach. Because of timely medical intervention, the patient managed to overcome the crisis and her reproductive organs were successfully preserved.

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.

Involvement of anticentromere antibody in interference with oocyte meiosis and embryo cleavage.

The aim of this study was to investigate the clinical significance of anticentromere antibody (ACA) among types of antinuclear antibody (ANA) in the properties of oocytes retrieved from infertile women. The rate of metaphase II oocytes or embryo cleavage was significantly decreased in patients with positive ACA compared with patients with negative ACA, suggesting that ACA is an essential marker for flawed oocytes in infertile women with any type of ANA.

Emerging strategies for ErbB ligand-based targeted therapy for cancer.

ErbB receptors are crucial for development and evolution and have been intensely pursued as targets for cancer therapeutics. Inhibiting the signaling activity of individual receptors in this family has advanced human cancer treatment. However, actual curative effects of the existing anti-ErbB therapeutics are still insufficient. A large percentage of patients who are initially responsive to ErbB receptor-targeted therapies later become resistant. Mechanisms responsible for tumor resistance to ErbB-targeted agents are as follows: many epidermal growth factor receptor (EGFR)- and HER2-targeted therapies cannot inhibit signaling through the ErbB receptor heterodimer, and anti-EGFR agents can suppress extracellular signal-related kinase (ERK) signal proliferation but not protein kinase B/Akt survival signals. ErbB ligand-based targeted therapy against HB-EGF or amphiregulin may overcome such obstacles. Here we discuss the efficacy of CRM197, a specific inhibitor of HB-EGF, and its possible clinical adaptation in combination with conventional chemotherapeutic agents in cancer therapy.

Apoptosis as a possible candidate mechanism for removal of tamoxifen-related endometrial cells with KRAS mutations.

BACKGROUND: Endometrial cell KRAS mutations are frequent in tamoxifen (TAM)-treated breast cancer patients. We previously demonstrated that most KRAS mutations disappeared after TAM cessation, suggesting the existence of a removal mechanism for endometrial cells with KRAS mutation. Here, the role of apoptosis in this mechanism was investigated. PATIENTS AND METHODS: DNA was extracted from frozen endometrial polyps of 31 TAM-treated breast cancer patients. Codon 12 mutations in KRAS were detected by enriched polymerase chain reaction enzyme-linked minisequence assay. Apoptosis was detected by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method and Ki-67 expression by immunohistochemistry. Relationships between KRAS mutations, the apoptosis index, and the Ki-67 index were determined. RESULTS: KRAS mutations were observed in 9 of these patients. There was no significant relationship between the Ki-67 index and KRAS mutation. However, the apoptosis index was significantly higher in polyps with KRAS mutation (p=0.002). CONCLUSION: Apoptosis may play an important role in removing TAM treatment-related endometrial cells with KRAS mutations.

HB-EGF inhibition in combination with various anticancer agents enhances its antitumor effects in gastric cancer.

Advanced gastric cancer (GC) is one of the most lethal malignancies. Although many anticancer agents exist for the treatment of GC, its prognosis remains extremely poor. Therefore, further development of targeted therapies is required for patients with GC. To assess the role of heparin-binding epidermal growth factor-like growth factor (HB-EGF) as a target for GC therapy, the expression of EGF receptor ligands in GC cell lines, and the antitumor effects of an HB-EGF inhibitor (CRM197) as a single agent and in combination with other anticancer agents was assessed in GC cells. HB-EGF was the predominantly expressed ligand among EGF receptor ligands in all the cells. CRM197 induced significant cell apoptosis. Anticancer agents augmented the secretion of HB-EGF into the medium and simultaneously induced cell apoptosis. Combination of CRM197 with other anticancer agents significantly enhanced cell apoptosis. Additionally, co-administration of CRM197 and paclitaxel resulted in synergistic antitumor effects. These results suggested that HB-EGF is a rational target for GC therapy.

Change in cervical length in cases resulting in threatened preterm labor.

PURPOSE: To determine the predisposing changes in cervical length (CL) and the critical range of CL in which significant uterine contractions emerge resulting in threatened preterm labor (TPL). METHODS: Sixty-eight uncomplicated singleton pregnancies where the CL was <25 mm before 31 weeks were divided into cases with TPL (n = 23) or without (n = 45). CL and uterine contractions were monitored sequentially starting between 16 and 20 weeks. The gestational ages when a CL of <25 or <15 mm was first observed, the interval between these two measurements, and the CL value at TPL diagnosis were analyzed retrospectively. RESULTS: (1) The gestational ages when a CL of <25 and <15 mm was first detected were lower in the TPL group (25 (median); 18-30 (range) and 28; 25-33 weeks, respectively) than in the non-TPL group (27; 20-30 and 33; 26-35 weeks; P = 0.030 and P < 0.001). (2) The interval between the two measurements was shorter in the TPL group (2.5; 0-15 weeks) than in the non-TPL group (5.5; 0-13 weeks, P = 0.034). (3) The CL value at TPL diagnosis was 13 mm (median), ranging from 7 to 18 mm. CONCLUSION: Cases with early onset and subsequent rapid CL shortening before 31 weeks resulted in TPL when CL decreased below the range 7-18 mm.

Efficacy of ligand-based targeting for the EGF system in cancer.

Although drugs inhibiting ErbB receptors such as epidermal growth factor receptor (EGFR) and HER2 have been developed as anticancer agents targeting the EGF family, they are not effective for all types of cancer and instead target only certain types. We propose the following four main reasons for these observations: (i) although seven EGFR ligands exist, effective inhibition of specific EGFR ligands may occur because their expression levels differ in different malignancies; (ii) suppressing EGFR ligands inhibits aggregation of EGFR and other ErbB receptors and activation of ERK and Akt signals; (iii) EGFR ligands may have various combinations for signal transduction through the EGFR pathway and other receptor signals; and (iv) the intracellular C-terminals of EGFR ligands move into the nucleus and strongly regulate cell proliferation. In this review, we describe important implications for targeted cancer therapy against EGFR ligands and describe the current situation in the development of ligand-based therapies for cancer.

[Ureteral endometriosis in estrogen-treated postmenopausal woman].

A 56-year-old woman was admitted to our hospital due to gross hematuria. Cystoscopy and contrast-enhanced abdominal CT scan revealed a solid tumor at the right terminal ureter. She underwent transurethral resection of the right ureteral tumor. The ureteral tumor included proliferative endometrial stroma and glands under urothelial cells, and the histopathological diagnosis was ectopic endometriosis. Before the surgery, the patient had underwent hormone replacement therapy using estrogen patches to treat menopausal disorders, however, the dose of estrogen was five times higher than regular dose. Therefore, in this case, the serum level of estrogen was elevated over the normal value. Ectopic endometriosis is rare in urinary tract especially ureter. Furthermore, endometriosis is an uncommon disease in women during postmenopausal period. Our case suggests that an excessive hormone replacement therapy might cause endometriosis in postmenopausal women.

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.

The efficacy of preoperative hormonal therapy before laparoscopic cystectomy of ovarian endometriomas.

AIM: The aim of this study was to investigate the influence of preoperative hormonal therapy before laparoscopic cystectomy of ovarian endometriomas. We identified differences in follicle loss and surgical difficulties with or without preoperative hormonal therapy. METHODS: Ninety-six patients with ovarian endometrioma underwent a laparoscopic cystectomy. Patients were divided into three groups: control group A (53 patients, 57 cysts) with no preoperative hormonal therapy; group B (34 patients, 40 cysts) who received gonadotropin-releasing hormone agonist therapy; and group C (9 patients, 11 cysts) who received danazol therapy before surgery. The medical and videotape records of all patients were retrospectively reviewed. The specimens of endometriomas were histologically evaluated. RESULTS: Mean diameters of endometriomas before hormonal therapy in groups B and C were significantly greater than those in the control group. There were no significant differences in the following: the mean diameter of removed cysts, the revised-American Society of Reproductive Medicine scores, the number of capsules containing follicle(s), and the mean number of follicles attached to a cyst. However, the number of capsules showing fibrosis significantly increased in the preoperative hormonal therapy groups (P < 0.001). Furthermore, the mean operation time of the preoperative hormonal groups was significantly longer than that of the control group (P < 0.01, P < 0.001). CONCLUSION: Our data suggested that preoperative hormonal therapy reduced the size of endometriomas. However, with similar revised-American Society of Reproductive Medicine scores, preoperative hormonal therapy did not contribute to the reduction of the loss of ovarian follicles. Fibrosis resulting from hormonal therapy appears to be responsible for these observations.