[A case of emergency operation for pedicle torsion of ovarian tumor during bevacizumab therapy].

We report a case of torsion of ovarian carcinoma pedicle during chemotherapy with bevacizumab and FOLFIRI. A 47-year-old-female who had undergone laparoscopic-assisted sigmoidectomy for sigmoid cancer with multiple metastatic liver tumors, was treated with 3 courses of FOLFOX4 and 9 courses of FOLFIRI. Bevacizumab (BV) was started to administer with FOLFIRI since a CT scan had revealed that metastatic liver tumors had enlarged . Four days after final administration of BV (13th course), she presented lower abdominal pain. CT scan showed an enlarged right ovary sized 20 cm, but no findings suggesting bowel perforation; therefore we assumed her pain was due to enlarged ovarian tumor, which might be metastasis from sigmoid cancer. Increased oral morphine could not relieve her pain. Then, along with sufficient information about the risks of the operation caused by BV, we proposed an operation to remove her enlarged ovary. Day 16 after final administration of BV, we undertook a laparotomy by lower median incision, and found a twisted right adnex with large necrotic ovarian tumor. Pathologists diagnosed the ovarian tumor as primary ovarian cancer. No complication related to BV occurred, and she was moved to the ward for internal medicine 13 days after operation.

A case of primary serous papillary carcinoma with unusual clinical presentation: distant lympho nodes metastasis without peritoneal dissemination.

CASE REPORT: A 50-year-old woman with primary serous papillary carcinoma (PSPC) of peritoneal origin located in the posterior uterine serosa and cul-de-sac without peritoneal dissemination. No peritoneal dissemination was detected but the tumor metastasized to para-aortic and supraclavicular lymph nodes. After first chemotherapy course, pericardial effusion occurred. A pericardiectomy was performed to prevent cardiac failure. Subsequent chemotherapy with paclitaxel and carboplatin was effective against this tumor CONCLUSION: In general, a typical type of PSPC usually develops distant metastasis with diffuse peritoneal dissemination; the present case shows unusual clinical behavior.

[Remarkably reduced tumor marker SCC levels by combined chemotherapy of paclitaxel and S-1 in two cases of advanced cervical cancer].

Case 1: A-35-year-old woman was diagnosed as cervical cancer Stage IIIb. When admitted to the hospital, her tumor marker SCC level was 50 ng/mL. Concurrent chemoradiation therapy was started on November, 2005. The SCC level was reduced by 0.9 ng/mL in February, 2006. In April, tumor recurrence was found by PET, and chemotherapy was restarted, but the SCC level was increased. In September, paclitaxel/S-1 therapy was performed, and the tumor markers were again reduced remarkably (SCC 9.8--> 1.3 ng/mL). Case 2: A-78-year-old woman was diagnosed as cervical cancer Stage IIIb. In August, 2004, concurrent chemoradiation therapy was started, and tumor markers were reduced (SCC 25.4--> 1.8 ng/mL). However, tumor markers were increased soon after the therapy. Chemotherapy was started, but it could not be maintained because of the side effects. In April, 2006, paclitaxel/S-1 therapy was performed, and the tumor markers were reduced remarkably (SCC 120--> 10 ng/mL). However, that therapy could also not be maintained because of the side effect. In July, she died of the cancer.

Differential activation of the luteinizing hormone beta-subunit promoter by activin and gonadotropin-releasing hormone: a role for the mitogen-activated protein kinase signaling pathway in LbetaT2 gonadotrophs.

LH consists of alpha- and beta-subunits, and synthesis of the beta-subunit has been reported to be the rate-limiting step in LH production. In this study, we found that activin A increased both the LHbeta mRNA level and LH content in cells of the gonadotroph cell line, LbetaT2. We next examined the effects of activin A and GnRH on LHbeta promoter activity by reporter gene assay and compared the signal transduction pathways. Activin A and GnRH activated the LHbeta promoter, and the response to a combination of activin A and GnRH was higher than that to activin A or GnRH alone. The effects of activin A and GnRH were specifically inhibited by inhibin-like peptide and antide, a GnRH antagonist, respectively. The activation of the LHbeta promoter by GnRH was inhibited by PD098059 and U0126, MAP kinase kinase (MEK) inhibitors. In contrast, these protein kinase inhibitors did not inhibit the activin A-induced activation. GnRH, but not activin A, activated MAP kinase in LbetaT2 cells. Overexpression of constitutively active MEK1 or MEK kinase activated both MAP kinase and the LHbeta promoter. Furthermore, GnRH, but not activin A, strongly induced SRE-mediated transcription, a known target of the MAP kinase pathway. These results suggest that GnRH activates the LHbeta promoter via the MAP kinase pathway and that activin A-induced activation of the LHbeta promoter is independent of the MAP kinase pathway.

Characterization of alphaT3-1 cells stably transfected with luteininzing hormone beta-subunit complementary deoxyribonucleic acid.

Luteinizing hormone (LH) consists of alpha- and beta-subunits, and synthesis and secretion of LH are regulated by gonadotropin-releasing hormone (GnRH). In order to examine the molecular mechanisms by which GnRH regulates LH secretion, we transfected alphaT3-1 cells with rat LHbeta-subunit cDNA under the control of a constitutive promoter and established a stable cell line of LH2 cells which secreted LH in response to GnRH. Pulsatile and continuous GnRH pretreatments increased gene expression of the alpha-subunit and synthesis of LH, and enhanced the LH secretion by brief treatments with GnRH and 56 mM KCl. The LH secretions were partially blocked by elimination of extracellular Ca2+. GnRH-induced LH secretion was completely inhibited by calphostin C (a protein kinase C inhibitor) and 1 microM wortmannin. In contrast to the GnRH induction, high K+-induced LH secretion was inhibited by KN93, a Ca2+/calmodulin-dependent protein kinase II inhibitor, as well as by 1 microM wortmannin. We also confirmed that activation of cAMP-pathway induced LH secretion, but activation of mitogen-activated protein (MAP) kinase pathway was not involved in LH secretion. These results suggest that GnRH directly regulates LH secretion as well as LHbeta-subunit synthesis, and that LH2 cells are a useful model for the study of LH secretion induced by several secretagogues.

Influence of missense mutation and silent mutation of LHbeta-subunit gene in Japanese patients with ovulatory disorders.

The frequency of variant LHbeta containing two point mutations (T(986)-C and T(1008)-C) and its relationship to reproductive disorders differ widely between ethnic groups. In a Japanese population, variant luteinizing hormone (LH) correlates with ovulatory disorders. Here we examined the relationship between two missense mutations and five silent mutations (C(894)-T, G(1018)-C, C(1036)-A, C(1098)-T and C(1423)-T) in the LHbeta gene, and ovulatory disorders. We studied 43 patients with ovulatory disorders, 79 patients with normal ovulatory cycles, and 23 healthy men who agreed to join our DNA analysis. PCR-amplified LHbeta-subunit gene sequences were compared with a base sequence of wild-type LH reported after direct sequencing. The highest frequency (0.945) of novel allele was observed at the position of the C(1036)-A transition. No homozygotes for wild-type LHbeta (C(1036)) were identified. The frequency of novel allele in patients with polycystic ovary syndrome, endometriosis, premature ovarian failure and luteal insufficiency was significantly different from that of healthy women. The frequencies of novel alleles (C(894)-T, C(1098)-T and C(1423)-T) in patients with ovulatory disorders were significantly higher than those with normal ovulatory cycles. The mean incidence of point mutation in patients with ovulatory disorders was higher than in those with normal ovulatory cycles. Among patients with variant LH, five silent mutations were identified in 87.5% of patients with ovulatory disorders, whereas only a few silent mutations were identified in patients with normal ovulatory cycles. In a Japanese population, five silent mutations of variant LH could have influenced two missense mutations and/or other unknown missense mutations, causing ovulatory disorders.

Regulation of gonadotropin alpha subunit gene expression by dopamine D(2) receptor agonist in clonal mouse gonadotroph alphaT3-1 cells.

Pituitary prolactin biosynthesis is negatively regulated by hypothalamic dopamine through D(2) receptors in pituitary lactotrophs, but little is known about the direct effect of dopamine on gonadotrophs. In this study, the clonal gonadotroph-derived cell line, alphaT3-1, was used to examine whether gene expression of the pituitary gonadotropin alpha subunit, stimulated with GnRH or pituitary adenylate cyclase-activating polypeptide (PACAP), was controlled by dopamine D(2) receptor. Western blotting and reverse transcription-polymerase chain reaction analysis demonstrated the presence of dopamine D(2) receptors in alphaT3-1 cells. Both GnRH and PACAP increased alpha subunit gene expression. GnRH-induced alpha subunit gene expression was not affected by quinpirol, a specific dopamine D(2) receptor agonist. In contrast, PACAP-induced gene expression was significantly lower in the presence of quinpirol. The roles of extracellular signal-regulated kinase (ERK) and cAMP in the expression of the alpha subunit gene were examined. GnRH activated ERK, but PACAP did not, and the activation was not inhibited by quinpirol. GnRH-induced alpha subunit gene expression was completely inhibited by an ERK inhibitor, PD098059. Cyclic AMP accumulation in alphaT3-1 cells was increased by treatment with PACAP, and quinpirol inhibited this effect. GnRH did not affect cAMP production in these cells. These results suggest that in alphaT3-1 cells, dopamine D(2) receptors negatively regulate pituitary alpha subunit gene expression in association with the cAMP-dependent pathway, but not with the ERK pathway.

Differential regulation of pituitary hormone secretion and gene expression by thyrotropin-releasing hormone. A role for mitogen-activated protein kinase signaling cascade in rat pituitary GH3 cells.

We examined the possible involvement of mitogen-activated protein (MAP) kinase activation in the secretory process and gene expression of prolactin and growth hormone. Thyrotropin-releasing hormone (TRH) rapidly stimulated the secretion of both prolactin and growth hormone from GH3 cells. Secretion induced by TRH was not inhibited by 50 microM PD098059, but was completely inhibited by 1 microM wortmannin and 10 microM KN93, suggesting that MAP kinase does not mediate the secretory process. Stimulation of GH3 cells with TRH significantly increased the mRNA level of prolactin, whereas expression of growth hormone mRNA was largely attenuated. The increase in prolactin mRNA stimulated by TRH was inhibited by addition of PD098059, and the decrease in growth hormone mRNA was also inhibited by PD098059. Transfection of the cells with a pFC-MEKK vector (a constitutively active MAP kinase kinase kinase), significantly increased the synthesis of prolactin and decreased the synthesis of growth hormone. These data taken together indicate that MAP kinase mediates TRH-induced regulation of prolactin and growth hormone gene expression. Reporter gene assays showed that prolactin promoter activity was increased by TRH and was completely inhibited by addition of PD098059, but that the promoter activity of growth hormone was unchanged by TRH. These results suggest that TRH stimulates both prolactin and growth hormone secretion, but that the gene expressions of prolactin and growth hormone are differentially regulated by TRH and are mediated by different mechanisms.