Kimio Mizuno

Effects of 17β-estradiol and progesterone on growth-factor-induced proliferation and migration in human female aortic smooth muscle cells in vitro

Objective: The significantly low prevalence of atherosclerotic cardiovascular disease in premenopausal women has been noted, and postmenopausal hormone replacement therapy (HRT) is associated with protection from this disease. The aim of this study was to investigate the effects of estrogen and progestin on growth-factor-induced vascular smooth muscle cell (VSMC) proliferation and migration in vitro. Methods: Two cell lines of human female aortic smooth muscle cells (AOSMCs) were used for the study. DNA synthesis was evaluated by [3H]thymidine incorporation into cells. Migration assay was performed using modified Boyden chambers. Results: The presence of estrogen receptors was determined by Western and Northern blot analyses. [3H]Thymidine incorporation into AOSMCs was induced by 10 ng/ml EGF alone, the combination of 10 ng/ml EGF with 2 ng/ml b-FGF-induced and 1 nM PDGF-BB alone. Migration of AOSMCs was induced by PDGF-BB. Since 17β-estradiol (E2, 10−9 M~ 10−6 M) inhibited the [3H]thymidine incorporation into AOSMCs stimulated by above mitogens and the 1 nM PDGF-BB-induced AOSMC migration in a dose-dependent manner, estrone (E1), estriol (E3) and progesterone (P) had no significant effects. The combination of P (10−9 M ~ 10−6 M) did not show any effect on these inhibitory effects of 10−7 M E2. Preincubation of AOSMCs with the anti-estrogenic agent, tamoxifen (10−6 M), significantly antagonized these inhibitory effects of 10−7 M E2. Conclusions: These findings suggest that the inhibitory effect of E2 on VSMC proliferation and migration might be one of the factors involved in the decreased incidence of atherosclerotic cardiovascular disease in premenopausal women and postmenopausal HRT, and P might not affect these estrogenic responses.

Prognostic factors in yolk sac tumors of the ovary. A clinicopathologic analysis of 29 cases

Twenty‐nine ovarian cancer patients with yolk sac tumors and germ cell tumors with yolk sac tissue as a component of their disease (16 endodermal sinus tumor, 11 mixed germ cell tumors, one embryonal carcinoma, and one polyembryoma) were treated with cytoreductive surgery and combination chemotherapy. Prognostic factors were investigated in this group. Patients with Stage I disease had a more favorable prognosis (P < 0.003) than those with Stages II and IV disease. The difference in prognosis was significant in cases where residual tumor was absent (P < 0.003) and in cases where ascites was either absent or less than 100 ml in volume (P < 0.05). Endodermal sinus tumor with either an intestinal (P < 0.05) or microcystic pattern (P < 0.01) was more common in survivors than in those who died. The age, preoperative serum alpha‐fetoprotein level, maximum tumor size, and tumor weight had no significant correlation with prognosis. In advanced cases, chemotherapy regimens including cisplatin gave better results than those containing vincristine, dactinomycin, and cyclophosphamide (P < 0.05). The optimal treatment of yolk sac tumors or tumors with yolk sac tissue as a component of the ovary is discussed in light of these results.

Characterization of extracellular matrix-degrading proteinase and its inhibitor in gynecologic cancer tissues with clinically different metastatic form

Background. The authors conducted a comparison study of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase‐1 (TIMP‐1) activities in clinically different metastatic types of ovarian cancer, cervical cancer, and endometrial cancer tissues.

Effects of 17β-estradiol and progesterone on migration of human monocytic THP-1 cells stimulated by minimally oxidized low-density lipoprotein in vitro

OBJECTIVE Many epidemiological studies have shown that postmenopausal hormone replacement therapy (HRT) has a beneficial effect on atherosclerotic cardiovascular disease. The aim of this study was to investigate the effects of estrogen and progestin on the migration of monocytes induced by minimally oxidized low-density lipoprotein (m-ox-LDL) in vitro. METHODS Human monocytic THP-1 cells were used for the study. Migration assay was performed using a modified Boyden chamber. RESULTS The presence of estrogen receptors was determined in THP-1 cells by Western and Northern blot analysis. Although native LDL had no significant effects on the migration of THP-1 cells, m-ox-LDL increased the migration of THP-1 cells in a dose-dependent manner. Although 17 beta-estradiol (E2, 10(-9)-10(-6) M) inhibited the 10 micrograms/ml-induced migration of THP-1 cells in a dose-dependent manner, estrone (E1), estriol (E3) and progesterone (P) had no significant effects. The combination of P (10(-9)-10(-6) M) did not show any effect on the inhibitory effect of 10(-7) M E2. Preincubation of THP-1 cells with the anti-estrogenic agent, tamoxifen (10(-6) M), significantly antagonized the inhibitory effect of 10(-7) M E2. m-ox-LDL stimulated MCP-1 secretion from THP-1 cells, which was reduced by E2. Anti-human MCP-1 neutralizing antibody inhibited the migration of THP-1 cells stimulated by m-ox-LDL. E2 also inhibited the 10 ng/ml MCP-1-induced migration of THP-1 cells in a dose-dependent manner. CONCLUSIONS These findings suggest that the inhibitory effect of E2 on the migration of monocytes might be one of the factors involved in the decreased incidence of atherosclerotic cardiovascular disease in premenopausal women and postmenopausal HRT.

Survival impact of capsule rupture in stage I clear cell carcinoma of the ovary in comparison with other histological types

OBJECTIVE We analyzed a large number of stage I clear cell carcinoma of the ovary (CCC) patients to estimate the survival impact of the capsule status in stage I CCC patients, particularly in comparison with non-CCC patients. METHODS Clinicopathologic data on 564 patients with stage I epithelial ovarian cancer (EOC) collected under the central pathological review system were subjected to uni- and multivariable analyses to evaluate the disease-free survival (DFS) and overall survival (OS). RESULTS There was no significant difference in both the OS and DFS of CCC patients between IA and IC(ir) (intraoperative capsule rupture) {IA vs. IC(ir); OS: P=0.1402, DFS: P=0.2701}. In contrast, CCC patients at IC(non-ir) {IC excluding for IC(ir), such as preoperative capsule rupture, positive ascites/washing, and surface involvement} showed a poorer OS and DFS than those at IC(ir), or those at the corresponding stage in non-CCC. In multivariable analysis, the capsule status was an independent prognostic factor of a poor OS and DFS {OS: HR, 2.832; 95% CI 1.156-6.938; P=0.023; DFS: HR, 4.327; 95% CI, 1.937-9.667; P=0.0004)} {In contrast, non-CCC: N.S. (OS/DFS)}. Furthermore, in CCC patients, intraperitoneal recurrences were more frequently observed in IC(non-ir) CCC than IA or IC(ir) CCC (P=0.0083) {In contrast, non-CCC: N.S.}. CONCLUSION This study suggests that CCC patients other than those with intraoperative capsule rupture show a considerable risk for mortality despite adjuvant chemotherapy.

Is there any possibility of fertility-sparing surgery in patients with clear-cell carcinoma of the ovary?

BACKGROUND In epithelial ovarian cancer (EOC), fertility-sparing surgery (FSS) has mainly been chosen for stage IA disease. The purpose of this study was to clarify the clinical outcome of patients with clear-cell carcinoma of the ovary (CCC) who would usually undergo radical surgery. CASES After a central pathological review and search of the medical records from multiple institutions between 1988 and 2005, a total of 10 CCC patients treated with FSS were retrospectively evaluated in the current study. The mean age was 35.9 years (range: 32-39 years). The median follow-up time was 35.4 months (range: 21.7-153.2 months). The stage was IA in 4 patients, and IC in 6 patients [IC(b) in 5 patients, and IC(2) in one]. Nine patients received adjuvant chemotherapy. Nine patients were alive and one patient with stage IC(2) died of the disease at a follow-up time of 36.8 months. Five pregnancies were observed in 4 patients. CONCLUSIONS Although there is no worldwide criterion for FSS in CCC patients at present, it seems that, in selected patients, this surgical approach could be adopted. This should be investigated by additional studies in a larger series.

Clinical value of a new serum tumor marker, CA125 II, in gynecologic disease : comparison with CA125

CA125 II, an improved version of the conventional CA125 was compared with CA125 as to which was more useful in gynecologic disease. In the diagnosis of tumors around the adnexal field (primary epithelial ovarian cancer, metastatic ovarian cancer, benign ovarian tumor and endometrial cyst), CA125 II showed the same sensitivity and specificity as CA125. CA125 II also has high simultaneous reproducibility in the low concentration area. The examination by the receiver operating characteristic curve revealed that CA125 II has higher precision than that of CA125 when it is used for the screening test. In conclusion, CA125 II is a better tumor marker than conventional CA125.

Fertility-sparing surgery in young women with mucinous adenocarcinoma of the ovary

OBJECTIVES The purpose of this study was to clarify the clinical outcome of patients with stage IA mucinous epithelial ovarian cancer (mEOC) treated with fertility-sparing surgery (FSS). METHODS After a central pathological review and search of the medical records from multiple institutions, a total of 148 stage I mEOC patients were retrospectively evaluated in the current study. All mEOC patients were divided into three groups: group A (FSS; age, 40≥); groups B and C {radical surgery; age, 40≥ (B); 40< (C)}. Survival analysis was performed among these three groups using Kaplan-Meier methods. RESULTS The median follow-up time of all mEOC patients was 71.6 (4.8-448.3) months. Among the 41 patients in group A, 27 patients (65.9%) had IA disease, and 14 (34.1%) had IC disease. Five-year overall survival (OS) and disease-free survival (DFS) rates of patients in the groups were as follows: group A, 97.3% (OS)/90.5% (DFS); group B, 94.4% (OS)/94.4% (DFS); group C; 97.3% (OS)/89.3% (DFS). Collectively, there was no significant difference in OS or DFS among these groups even though they were stratified to each substage (IA/IC) (OS, P=0.180; DFS, P=0.445, respectively). Furthermore, in multivariate analyses, the surgical procedure was not an independent prognostic factor for either OS or DFS (OS, HR: 0.340, 95% CI: 0.034-3.775, P=0.352; DFS, HR: 0.660, 95% CI: 0.142-3.070, P=0.596). CONCLUSIONS Patients with stage I mEOC treated with FSS did not necessarily show a poorer prognosis than those receiving radical surgery.

Transfection of human cytochrome P-450 reductase cDNA and its effect on the sensitivity to toxins

NADPH-cytochrome P-450 reductase (CYPR; EC 1.6.2.4) is an essential enzyme for the catalytic function of cytochromes and is also regarded as being implicated in drug activation. To examine whether CYPR is directly involved in the drug metabolism, a cDNA encoding human CYPR was stably transfected into Chinese hamster ovary cells. Three clonal cell lines were identified which expressed increased levels of CYPR activity (9.3- to 11.2-fold). Western blot analysis indicated that CYPR-transfectant cells contained markedly elevated levels of CYPR protein, while wild-type Chinese hamster ovary cells contained low levels. They showed 1.8- to 3.3-fold higher sensitivity to Adriamycin, 2.1- to 3.0-fold higher sensitivity to mitomycin C, and 2.9- to 4.3-fold higher sensitivity to paraquat than wild-type cells. We also studied other common-use anticancer agents: vinblastine, vincristine, VP-16, and cisplatin, but there were no differences observed in the sensitivity. This report provides the first evidence that transfection of human CYPR into mammalian cells is concerned in the sensitivity to some drugs.

Recurrence of epithelial ovarian carcinoma after clinical remission.

One hundred and eighty-eight patients with epithelial ovarian carcinoma were treated with primary cytoreductive surgery and subsequent combination chemotherapy. The first recurrent findings such as sites and disease-free interval were analyzed in 141 patients who were clinically remitted 6 months after operation or chemotherapy. Fifty-seven cases had a recurrence. Five-year disease-free survival rates were 75, 72, 29, and 0% in stage I, II, III, and IV, respectively. Twenty-one of 22 patients with > 2 cm maximum residual tumor died, although they once achieved clinical remission. Significant differences were observed between histologic types, and the disease-free survival rate was lowest for serous cystadenocarcinoma. Nine of 15 stage IV patients with serous histology experienced remission, but none of the 8 in stage IV with other histologies did so, suggesting that serous adenocarcinoma is sensitive to chemotherapy and conducive to clinical remission. However, all stage IV patients in remission encountered a recurrence. Intra-abdominal cavity and lymph node were frequently the initial recurrent sites (38 and 27%, respectively). On the other hand, the incidence of distant recurrence was as high as 27%, and 8 of 16 cases with distant recurrence were stage I. Survival time after recurrence was not different among initial sites of recurrence and mean survival time was 15 months.