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Naohiko Umesaki

Publications and source records attributed to Naohiko Umesaki.

At least 19 recordsLinked to original sources
PubMed2006

Increased type III and V collagen expression in human corpora lutea in early pregnancy.

OBJECTIVE: To investigate the possible involvement of collagen in the characteristic structure and function of human corpora lutea (CL), type V collagen expression was determined in the CL tissues during the menstrual cycle and early pregnancy. DESIGN: In vitro experiment. SETTING: Department of Obstetrics and Gynecology, Wakayama Medical University, Wakayama, Japan. PATIENT(S): Regulatory cycling women and pregnant women with ovarian tumor and ectopic pregnancy who underwent adnexectomy. INTERVENTION(S): Composition of the various types of collagen in human CL was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. MAIN OUTCOME MEASURE(S): Expression of type V collagen. RESULT(S): The ratios of type III to type I collagen and the ratios of type V to type I collagen in the CL tissues were significantly increased in early pregnancy compared with those in the menstrual cycle. CONCLUSION(S): These results suggest that alterations in composition of collagen might play an important role in determining the physiology and structure of the CL during the menstrual cycle and early pregnancy.

Adult↗
PubMed2006

Impaired postnatal development in C/EBPbeta-deficient mice.

Tail DNA genotyping of fetal and neonatal mice from C/EBPbeta heterozygous parents was performed to determine whether the decreased number of surviving C/EBPbeta mutants was caused by prenatal or postnatal death. Eighty-four 3-week-old mice born of heterozygous parents had significantly lower numbers of C/EBPbeta-deficient offspring than the expected Mendelian ratio (29.8%+/+, 65.5%+/-, 4.76%-/-, P<0.05). The genotypes of 72 fetal mice from intercrossed heterozygotes showed approximately the expected 1:2:1 Mendelian ratio (18.1% +/+, 52.8% +/-, 29.2% -/-, P>0.05). No difference in the proportions by sex could be detected in these perinates. This data indicates that C/EBPbeta-deficient mice have unknown lethal problems between the embryonic stage and weaning.

Animals↗
PubMed2006

Immunohistochemical analysis of collagen expression in human corpora lutea during the menstrual cycle and early pregnancy.

OBJECTIVE: To investigate the characteristic structure and function of human corpora lutea (CL), various types of collagen expression were determined in the CL tissues during the menstrual cycle and early pregnancy. DESIGN: In vitro experiment. SETTING: Department of obstetrics and gynecology at a medical university. PATIENT(S): Regulatory cycling women and pregnant women with ovarian tumor and ectopic pregnancy who underwent adnexectomy. INTERVENTION(S): Immunohistochemistry for human type I, III, and IV collagen with specific monoclonal antibodies was used for analysis. MAIN OUTCOME MEASURE(S): Expression of type I, III, and IV collagen. RESULT(S): Immunohistochemical staining for type I and III collagen revealed intense staining of the CL stroma during early pregnancy, as compared with those in the menstrual cycle. Moreover, pericellular intense immunostaining for type IV collagen was observed around the luteal cells, especially luteal granulosa cells, of early pregnancy. CONCLUSION(S): These results suggest that alterations in distribution of collagen might play an important role in determining the physiology and structure of the CL during the menstrual cycle and early pregnancy.

Adult↗
PubMed2006

A novel mechanism for acquired cisplatin-resistance: suppressed translation of death-associated protein kinase mRNA is insensitive to 5-aza-2'-deoxycitidine and trichostatin in cisplatin-resistant cervical squamous cancer cells.

The molecular mechanism for cisplatin (CDDP)-resistance of cancer cells has not yet been clarified, despite extensive studies. Here, we investigated whether death-associated protein (DAP) kinase, an apoptosis modulator, was involved in CDDP-resistance by examining the ME180 human cervical squamous cancer cell line and 6 monoclonal ME180-derived CDDP-resistant subclones. Co-treatment with CDDP and 5-aza-2'-deoxycytidine (5-aza-CdR), a demethylating agent, significantly enhanced the CDDP-sensitivities of the parent cells and CDDP-resistant subclones. Subsequent removal of 5-aza-CdR rapidly reversed the CDDP-sensitivity of the CDDP-resistant subclones to their original levels, whereas the parent cells retained the enhanced CDDP-sensitivity for at least 24 h. Quantitative RT-PCR revealed that the CDDP-resistant subclones expressed higher DNA methyltransferase (DNMT) mRNA levels than the parent cells, suggesting that increased DNMT expressions easily restored the CDDP-resistance of the CDDP-resistant subclones following 5-aza-CdR removal. Although the parent cells showed hypermethylation in the DAP kinase promoter region, corresponding methylated bands were not detected in 2 of the 6 CDDP-resistant subclones by methylation-specific PCR. All 6 CDDP-resistant subclones expressed higher DAP kinase mRNA levels than the parent cells, as evaluated by quantitative RT-PCR. Although DAP kinase protein expression was strongly suppressed in the parent cells and CDDP-resistant subclones, 5-aza-CdR treatment of the parent cells dose-dependently stimulated the DAP kinase protein expression, and this was synergistically enhanced by inhibiting histone deacetylation via trichostatin treatment in addition to 5-aza-CdR. However, DAP kinase protein expression in the CDDP-resistant subclones was not stimulated by treatment with 5-aza-CdR and/or trichostatin. These results indicate that post-transcriptional translation of DAP kinase mRNA is strongly suppressed and insensitive to treatment with 5-aza-CdR and trichostatin in the CDDP-resistant subclones established from ME180 human cervical squamous cancer cells. This CDDP-resistance is accompanied by molecular changes that disturb the post-transcriptional translation of the DAP kinase mRNA, and these molecular changes are transiently restored by demethylation.

Antineoplastic Agents↗
PubMed2006

Immunohistochemical localization of inhibin and activin subunits, activin receptors, and Smads in ovarian clear cell adenocarcinoma.

Our purpose was to examine the immunolocalization of inhibin, activin and the activin signaling system in ovarian clear cell adenocarcinoma tissue. Tissue samples of ovarian clear cell adenocarcinoma were collected from ten women. The patients' ages ranged from 41 to 71 years (mean +/- standard deviation; 53.4+/-9.9). Their surgicopathological staging was based on the criteria of the International Federation of Gynecology and Obstetrics (FIGO); there were 5 patients with stage I, 3 with stage II, and 2 with stage III. We examined the immunohistochemical localization of inhibin/activin alpha-subunit, betaA-subunit, activin A, and activin receptor types IA, IB, IIA, IIB, Smad2, Smad3 and Smad4 using an avidin-biotin peroxidase complex technique. With the antibodies against the betaA-subunit, and activin A, we observed positive immunoreactive staining in the cytoplasm of clear cell adenocarcinoma, whereas we did not observe any staining for the alpha-subunit of inhibin. We observed positive immunoreactive staining in the cell cytoplasm and surface with the antibodies against the activin receptors, types IA, IB, IIA and IIB. We observed positive immunoreactive staining in the cytoplasm and nucleus with the antibodies against the Smad2, Smad3 and Smad4. It is suggested that activin A, not inhibins, might be secreted by ovarian clear cell adenocarcinoma, and that an activin signal transduction pathway might exist in ovarian clear cell adenocarcinoma.

Activin Receptors↗
PubMed2006

Optimal combination chemotherapy and chemoradiotherapy with etoposide for advanced cervical squamous cancer cells in vitro.

Patients with unresectable advanced carcinoma of the uterine cervix are usually treated with chemotherapy or chemoradiotherapy. In the present study, the optimal administration protocol for etoposide in chemotherapy and chemoradiotherapy for advanced cervical cancer patients was investigated in vitro using the radio-sensitive and anticancer drug-sensitive human cervical squamous cell carcinoma cell line ME180. Therapeutic doses of concurrent irradiation reduced the cellular etoposide sensitivity in a dose-dependent manner, while postirradiation-surviving subclones established from repeatedly irradiated ME180 cells showed significantly higher etoposide sensitivities than the non-irradiated parent cells. Of the 6 monoclonal etoposide-resistant subclones established from ME180 cells, 5 were significantly radioresistant. Although the etoposide-resistant subclones were also significantly resistant to other anticancer drugs, such as cisplatin, carboplatin, nedaplatin, pirarubicin, paclitaxel and docetaxel, they were more sensitive to 5-fluorouracil, mitomycin C and SN38 than the parent cells. Flow cytometric analyses revealed that the etoposide-resistant subclones showed significantly increased cell surface expression of CD40 compared to the parent cells, which expressed undetectable levels of CD40. However, the expression of some integrin receptor subunits, such as CD29, CD49a and CD49f, was apparently reduced in the etoposide-resistant subclones. These results indicate that etoposide should be administered to advanced cervical squamous cancer patients after the completion of radiotherapy, rather than as a concurrent chemoradiotherapy. In order to kill surviving etoposide-resistant cancer cells more effectively, 5-fluorouracil, mitomycin C and irinotecan may be candidate combination drugs for use with etoposide. Differential expression of integrin receptors and CD40 may be involved in the acquisition of etoposide resistance by cervical squamous cancer cells.

Antigens, CD20↗
PubMed2006

The autocrine effect of activin A on human ovarian clear cell adenocarcinoma cells.

The functions of activin, a member of TGF-beta superfamily, in ovarian clear cell adenocarcinoma remain unsolved, although we recently found that inhibin betaA-subunit, activin A, activin receptor type IA, type IB, type IIA, type IIB, Smad2, Smad3 and Smad4 were localized in tumor cells of the ovarian clear cell adenocarcinoma tissue by immunohistochemistry. In the present study, in order to investigate the role of activin concerning cell growth in ovarian clear cell adenocarcinoma cells, we determined the production of activin A and inhibin A, and the expression of activin receptors and Smads using the human ovarian clear cell adenocarcinoma cell line JHOC-5. Moreover, we examined the effects of activin A on the activation of activin signaling pathway and on the proliferation in JHOC-5 cells. We detected a measurable amount of activin A in the culture medium of JHOC-5 cells, although inhibin A was not detected. The expression of activin receptor type IA, IB, IIA, IIB, Smad2, Smad3 and Smad4 was observed in JHOC-5 cells. Activin A induced a significant increase in proliferation of JHOC-5 cells compared with the untreated control. On the other hand, activin A did not affect the growth of JHOC-5 cells and no statistically significant difference was observed in the presence of follistatin which is a specific binding protein of activin. Phosphorylated Smad2, an activated form of Smad2, was detected both in treated JHOC-5 cells and in untreated cells by activin A. Activin A significantly increased the expression of phosphorylated Smad2 in JHOC-5 cells. Therefore, it is possible that activin has autocrine roles in tumor growth of ovarian clear cell adenocarcinoma cells.

Activin Receptors↗
PubMed2005

Mutational analysis of TP53 and p21 in familial and sporadic ovarian cancer in Japan.

OBJECTIVE: To investigate whether somatic mutations in cell cycle checkpoint genes, TP53 and p21, are involved in the development of ovarian cancer with or without BRCA1 germline mutation. METHODS: We analyzed somatic genetic alterations of TP53 and p21 in 46 ovarian cancer patients with BRCA1 germline mutations and 93 sporadic patients, using direct sequencing for the entire coding sequences in TP53 and p21. RESULTS: TP53 somatic mutations were detected in 25 of the 46 BRCA1 cases and 40 of the 93 sporadic cases (54.3% vs. 43.0%). In contrast, p21 somatic mutations were detected in 1 of the 46 BRCA1 cases and 2 of the 93 sporadic cases (2.2% vs. 2.2%). TP53 mutations in sporadic cases more frequently occurred in exons 6-11 than those in cases with germline BRCA1 mutations (84.4% vs. 56.3%: P = 0.013). The proportion of sporadic cases with TP53 mutations in non-serous tumors (e.g. endometrioid, clear cell, or mucinous) was significantly lower than that in serous tumors (18.5% vs. 53.0%: P = 0.0038). However, there was no significant difference between the proportion of BRCA1 cases with TP53 mutation in non-serous and in serous tumors (37.5% vs. 57.9%). CONCLUSIONS: Our results suggest that somatic mutation of TP53 plays less of a role in the carcinogenesis of sporadic non-serous tumors than in that of sporadic serous tumors or BRCA1-related tumors. Furthermore, p21 somatic mutation appears to be less involved in the development of ovarian cancer than TP53 somatic mutation.

Adult↗
PubMed2005

Mutational analysis of the estrogen receptor-alpha gene in familial ovarian cancer.

AIM: The genetic region of 6q25 containing the estrogen receptor-alpha (ER-alpha) gene is lost in a significant number of ovarian tumors. The aim of this study was to identify how inherited variation in the ER-alpha gene contributes to susceptibility to familial ovarian cancer. METHODS: DNA obtained from 18 cases of familial ovarian cancer without mutation of the BRCA1 and BRCA2 genes, 20 cases with BRCA1 mutation, 20 cases of sporadic ovarian cancer, and 19 controls were screened for mutations in the coding region of the ER-alpha gene using direct sequencing. RESULTS: Two germline missense variants at codons 307 [GCC(Ala)-->TCC(Ser)] and 347 [ACC(Thr)-->TCC(Ser)] were detected in two unrelated cases with BRCA1 mutation, but not in all other cases tested. Three polymorphisms in codon 10 [TCT-->TCC(Ser)], codon 325 [CCC-->CCG(Pro)], and codon 594 [ACA-->ACG(Thr)] were identified in this series, and a higher frequency of the allele TCC at codon 10 and a lower frequency of the allele CCG at codon 325 were observed in familial cases without BRCA1 mutation, compared with those in familial cases with BRCA1 mutation, in both the sporadic cases and in the controls. CONCLUSIONS: We could not detect pathogenic mutations of the ER-alpha gene in ovarian cancer cases without BRCA1 mutation. However, association analyses of two polymorphisms suggest that the ER-alpha gene or a gene located close to the ER-alpha locus might be related to susceptibility of familial ovarian cancer without BRCA1 mutation.

Adult↗
PubMed2005

Radiation reduces pirarubicin sensitivity in human cervical squamous cell carcinoma cells.

To find an effective protocol for chemoradiotherapy with pirarubicin hydrochloride (THP) for advanced cervical cancer patients, effects of irradiation on THP sensitivity were examined using the radiosensitive human cervical squamous cell carcinoma cell line ME180. Concurrent irradiation significantly reduced THP sensitivity, and this was further reduced when cells were treated with THP 8 h after irradiation. However, the THP sensitivity of cells treated with THP 8 h before irradiation was significantly enhanced. Four months after the first irradiation, 4 subclones of ME180 cells that survived repetitive irradiation demonstrated significantly higher THP sensitivity than non-irradiated parent cells. These results indicate that THP injections, more than several hours before irradiation or after completion of radiotherapy, might be better therapies than concurrent chemoradiotherapy with THP for cervical cancer.

Carcinoma, Squamous Cell↗
PubMed2005

Reduced radiosensitivity and increased CD40 expression in cyclophosphamide-resistant subclones established from human cervical squamous cell carcinoma cells.

To investigate the interaction between anticancer drug resistance and radioresistance in cervical cancer cells, 3 single cell-derived cyclophosphamide-resistant subclones were established from the drug- and radiosensitive human cervical squamous cell carcinoma cell line ME180 by chronic exposure cultures with 4-hydroxy-cyclophosphamide followed by limiting dilution. The established cyclophosphamide-resistant subclones were also radio- and multidrug-resistant to 7 other anticancer drugs. Flow cytometric analysis revealed significantly increased levels of CD40 expression on the 3 resistant subclones, whereas no CD40 expression was found on the parent ME180 cells. However, there were no changes in the expression levels of CD29, CD49a-CD49f or CD59 between the parent cells and resistant subclones. A recombinant human soluble CD40 ligand had no effect on the proliferation of the resistant subclones. Irradiation had no effect on the 4-hydroxy-cyclophosphamide sensitivity of the parent cells. These results indicate that the established cyclophosphamide-resistant subclones have impaired cell death signals, which are common to both drug- and radiation-induced apoptosis, and cyclophosphamide may not be an adequate drug for use in concurrent chemoradiotherapy. Furthermore, CD40 activation signals may be associated with the multidrug- and radioresistance in these cyclophosphamide-resistant subclones.

Antineoplastic Agents↗
PubMed2005

Radiation-induced cell death is independent of the apoptotic signals mediated by death-associated protein kinase in human cervical squamous cell carcinoma cells.

Death-associated protein kinase (DAPK) was originally identified as a positive mediator of interferon-gamma (IFNgamma)-induced apoptosis in cervical cancer cells, and interferons have been reported to enhance radiosensitivity in various types of squamous cell carcinoma. To examine whether DAPK can regulate cancer cell radiosensitivity, we investigated DAPK expression and radiosensitivity in human cancer cell lines, including the cervical squamous cell carcinoma cell line, ME180, which is both radiosensitive and IFNgamma-sensitive. Of the 5 human cancer cell lines examined, ME180 cells were the most radiosensitive, but their level of DAPK protein expression was undetectable by western blotting. A comparative study of ME180 cells with 2 other uterine cancer cell lines, HHUA and HOKUG, revealed no significant relationships between cellular radiosensitivity and DAPK protein expression or hypermethylation of the DAPK promoter CpG island. INFgamma dose-dependently inhibited ME180 cell proliferation, but did not induce any cell death. IFNgamma significantly enhanced the radiosensitivity of ME180 cells with a slight increase in DAPK protein expression, while irradiation significantly reduced their sensitivity to the growth-inhibitory signals of INFgamma. Analyses of 6 monoclonal cisplatin-resistant subclones established from ME180 cells revealed that all 6 were significantly more radioresistant than the parent ME180 cells without any change in the DAPK protein expression. These results indicate that DAPK does not regulate radiation-induced cell death and that it cannot be either a target molecule for radiotherapy with gene therapy or a prognostic marker for cervical cancer patients treated with radiotherapy.

Antineoplastic Agents↗
PubMed2005

Cell proliferation of bulky cervical squamous cell carcinoma is strongly associated with a paracrine tissue-specific growth factor(s).

To examine the possibility of a cervical squamous cell carcinoma-specific growth factor(s), previously suggested by clinical findings and in vitro culture experiments, we examined in vitro cultures and nude mouse transplantations of cervical squamous cell carcinoma cells obtained from a patient with a bulky cervical tumor without detectable distant metastases. The cancer cells did not proliferate without additional growth factors but remained viable in vitro. Fourteen weeks after transplantation of the tumor cells on their backs, 1 of the 3 nude mice developed large metastatic pelvic tumors without macroscopic metastatic lesions in their lungs or liver. When these pelvic tumor fragments were trans-planted onto the backs of other nude mice, ulcerated back tumors and larger metastatic pelvic tumors, but no macroscopic metastatic lesions in the lungs or liver, were observed. Histopathological examination of these pelvic tumors showed that all were the same squamous cell carcinoma as the primary tumor. These results indicate that cell proliferation of bulky cervical squamous cell carcinoma is strongly associated with a squamous cell carcinoma-specific growth factor(s) in a paracrine manner.

Animals↗
PubMed2005

Combination effects of irradiation and irinotecan on cervical squamous cell carcinoma cells in vitro.

Irinotecan HCl (CPT-11) has frequently been used in chemotherapy or concurrent chemoradiotherapy for patients with advanced cervical cancer, although an effective protocol for chemoradiotherapy with CPT-11 has not yet been established. Using the radiosensitive human cervical squamous cell carcinoma cell line ME180 and SN38, a major active metabolite of CPT-11, both the SN38 effects on radiosensitivity and irradiation effects on SN38 sensitivity were investigated to optimize the chemoradiotherapy protocol for CPT-11. SN38 had no effect on radiosensitivity, and irradiation did not affect SN38 sensitivity. Moreover, 3 of 4 post-irradiation surviving subclones obtained from repeatedly irradiated ME180 cells showed no significant changes in their SN38 sensitivities compared with the non-irradiated parent cells. On the other hand, all 7 SN38-resistant subclones established from ME180 cells showed strong reduction in their radiosensitivities. These results suggest that CPT-11 should be administered to advanced cervical cancer patients after, but not before, standard radiotherapy, and concurrent administration of SN38 with radiotherapy should be avoided in order to prevent severe adverse effects, such as watery diarrhea, which is the main adverse effect of pelvic radiotherapy or CPT-11.

Antineoplastic Agents, Phytogenic↗
PubMed2004

MR and ultrasound imaging of floating globules in mature ovarian cystic teratoma.

A rarely reported case of cystic teratoma with floating globules or fat balls is presented. Comparative imaging investigations of the floating globules in an ovarian cystic teratoma were performed using magnetic resonance and two- and three-dimensional ultrasound. Ultrasound imaging easily detected the floating globules, especially the 3-dimensional features showed almost the same findings that were observed macroscopically after surgery. Magnetic resonance thus clearly revealed the character of floating globules.

Female↗
PubMed2004

Phase II study of irinotecan combined with mitomycin-C for advanced or recurrent squamous cell carcinoma of the uterine cervix: the JGOG study.

OBJECTIVES: The efficacy and toxicity of combined therapy with irinotecan (CPT-11) plus mitomycin-C (MMC) were evaluated in patients with advanced or recurrent squamous cell carcinoma (SCC) of the uterine cervix. METHODS: CPT-11 (100 mg/m(2)) was administered on days 1, 8, and 15 by intravenous (iv) infusion over 90 min, while MMC (10 mg/m(2) iv) was given on day 1. This regimen was repeated every 28 days and at least two courses were given. RESULTS: Among 51 eligible patients (median age: 52 years; range: 25-72 years), 2 showed complete response (CR) and 24 showed PR, for an overall response rate (ORR) of 51.0% (95% confidence interval: 36.6-65.3%). In patients without prior chemotherapy, the ORR was 54.8% (38.7-70.2%). Twenty-five patients (Ib2:3, IIb:17, and IIIb:5) received this regimen as neoadjuvant chemotherapy and their ORR was 76% (54.9-90.6%). Twenty-two patients were able to undergo radical surgery after NAC. The major toxicity was neutropenia, which was grade 3-4 in 59% of the patients. Grade 3-4 thrombocytopenia and anemia were also seen in 26% of the patients each. The most common nonhematologic toxicity was diarrhea (grade 3-4 in 12%). CONCLUSION: CPT-11 combined with MMC can be effective against advanced or recurrent SCC of the uterine cervix.

Adult↗
PubMed2004

Reduced expression of death-associated protein kinase in human uterine and ovarian carcinoma cells.

The expression of the death-associated protein kinase (DAPK) protein and promoter methylation in 11 human uterine and ovarian carcinoma cell lines originally established from histopathologically-different carcinoma tissues were examined to investigate the relationship between DAPK and carcinogenesis in female reproductive tissues. The 11 cell lines included three cervical carcinomas, three endometrial carcinomas and five ovarian carcinomas. Western blot analysis showed no detectable expression of DAPK protein in 4 cell lines (ME180, HOKUG, MCAS and OVK-18) while moderate levels of DAPK protein were readily detected in normal human endometrium, and normal murine uterus and ovary. Methylation-specific PCR of the 11 cell lines revealed that 5 carcinoma cell lines (ME180, HOKUG, MCAS, OVK-18 and HEC-1) expressed hypermethylated promoters in the DAPK genes, while DAPK promoters in the other 6 carcinoma cell lines remained unmethylated. These results indicate that DAPK protein expression is reduced or silenced in some human uterine and ovarian carcinoma cells by methylation of the DAPK gene promoter region. Therefore, reduced DAPK expression and methylation of the DAPK promoter may be involved in carcinogenesis of human uterine and ovarian tissues.

Animals↗