Immunohistochemical expression of gonadotropin releasing hormone receptor in human breast carcinoma.

Gonadotropin releasing hormone (GnRH) analogs can cause regression of hormone-dependent breast carcinomas via the specific GnRH receptor (GnRH-R). In an attempt to obtain a better understanding of GnRH actions in human breast carcinoma, the expression of GnRH-R was examined immunohistochemically in 58 invasive ductal carcinomas and correlated with various clinicopathological parameters. GnRH-R was immunolocalized in the cytoplasm of carcinoma cells in 37 of 58 invasive ductal carcinoma cases (64%). Immunoreactivity for GnRH-R was also detected focally in the cytoplasm of morphologically normal glandular epithelia adjacent to the carcinoma. A significant correlation was observed between the immunohistochemical expression of GnRH-R and estrogen receptor labeling index (LI; P = 0.030) or progesterone receptor LI (P = 0.0074). There was a significant inverse correlation between GnRH-R immunoreactivity and Ki-67 LI (P = 0.012). No significant correlations were detected between GnRH-R and other clinicopathological parameters, including patient age, menopausal status, stage, tumor size, lymph node status, histological grade and prognosis. This study indicates that GnRH-R is widely distributed in human breast carcinoma cells and regulates GnRH actions locally. Breast carcinomas positive for GnRH-R maintain some hormonal regulatory mechanisms, and GnRH actions may lead to a low proliferative rate in human breast carcinoma.

5alpha-reductases in human breast carcinoma: possible modulator of in situ androgenic actions.

The expression of 5alpha-reductase types 1 and 2 was examined in human breast carcinoma using immunohistochemistry and RT-PCR. Immunoreactivity for 5alpha-reductase isozymes was also correlated with various clinicopathological parameters to examine possible local regulatory mechanisms of sex steroids, including progesterone and androgens, in human breast carcinoma tissues. Immunoreactivity for 5alpha-reductase type 1 was detected in the cytoplasm and possibly in the nuclear membrane of tumor cells in 35 of 60 invasive ductal carcinomas (58%), and type 2 signal was detected in 9 of these 60 cases (15%). The results from RT-PCR (n = 8) were consistent with those from immunohistochemistry. A significant positive correlation was detected between 5alpha-reductase type 1 immunoreactivity and androgen and progesterone receptor A or B labeling indexes, and immunoreactivities of 5alpha-reductase type 2, 17beta-hydroxysteroid dehydrogenase type 5, or 3beta-hydroxysteroid dehydrogenase, which recognizes both types I and II. An inverse correlation was detected between 5alpha-reductase type 1 immunoreactivity and tumor size, histological grade, or Ki-67 labeling index. 5alpha-Reductase type 2 immunoreactivity was significantly correlated with 17beta-hydroxysteroid dehydrogenase type 5 immunoreactivity, but not with other parameters. This study suggests that 5alpha-reductase type 1 is mainly expressed in human breast carcinoma, which may play an important role in the in situ production and actions of the potent androgen, 5alpha-dihydrotestosterone, including inhibition of cancer cell proliferation, in hormone-dependent human breast carcinoma.

Retinoid receptors in human breast carcinoma: possible modulators of in situ estrogen metabolism.

Retinoid receptors (retinoic acid (RARs) and retinoid X (RXRs) receptors) were immunolocalized in 32 human invasive ductal breast carcinomas. These findings were correlated with clinicopathological parameters to study their biological significance in breast carcinoma. Retinoid receptor immunoreactivity, except for RXRgamma, was detected in the nuclei of carcinoma cells. Percentage of positive cases were RARalpha; 81%, RARbeta; 6%, RARgamma; 28%, RXRalpha; 81%, and RXRbeta; 59%. A significant correlation was detected between RARalpha labeling index (LI), and RXRalpha LI (r = 0.667, p < 0.001). Results from immunoblotting performed in three cases were consistent with those of immunohistochemistry. There was a significant correlation between RARalpha LI and 17beta-hydroxysteroid dehydrogenase (17beta-HSD) type 1 immunoreactivity (p < 0.05). A significant correlation was also detected between RARalpha (r= 0.413, p = 0.019) or RXRalpha (r = 0.429, p = 0.014) LI, and estrogen receptor (ER) LI. In T-47D breast cancer cells, which express RARalpha, RXRalpha and ER, 17beta-HSD reductive activity increased 1.76-fold (p < 0.001), five days following treatment with 10 nM retinoic acid. These data suggest that retinoid receptors modulate various effects of retinoids, including estrogen metabolism in human breast carcinomas.

Stromal expression of fibroblast activation protein/seprase, a cell membrane serine proteinase and gelatinase, is associated with longer survival in patients with invasive ductal carcinoma of breast.

Fibroblast activation protein (FAP)/seprase is a serine integral membrane proteinase with gelatinase activity, which is expressed by activated fibroblasts in the stroma of various epithelial cancers, mesenchymal tumors and breast-cancer cells, as well as during wound repair. However, the pathophysiologic significance of its expression remains poorly understood. The present study was designed to reveal the impact of stromal expression of FAP/seprase on survival in human breast cancer. Immunohistochemical expression of FAP/seprase was restricted to stromal fibroblasts adjacent to tumor-cell nests but not cancer cells, which was confirmed by double-labeling immunohistochemistry. Clinicopathologic analysis revealed that more abundant FAP/seprase expression in 112 cases of invasive ductal carcinoma is associated with longer overall and disease-free survival. Multivariate analysis with other clinicopathologic factors demonstrated that FAP/seprase expression is an independent prognostic factor. The effect on the survival rate of FAP/seprase was also apparent in cases with lymph node metastasis. FAP/seprase expression is one of the manifestations of the stromal reaction (i.e., matrix turnover); thus, invasive ductal carcinomas with fewer stromal reactions expressing FAP/seprase may be more aggressive.

Progesterone receptor A and B isoforms in the human breast and its disorders.

Two different isoforms of progesterone receptor (PR), PRA and PRB, are expressed in target tissues at comparable levels. In this study, we first examined PRA and PRB immunoreactivity in human breast cancer and various intraductal proliferative epithelial lesions, and correlated these findings with clinicopathologic parameters. We then examined mRNA expression of PRA and PRB in six cases of invasive ductal carcinoma using RT-PCR. Immunoreactivity for both PRA and PRB was positive in the great majority of proliferative disease without atypia (PDWA) (85% for PRA and 96% for PRB) and atypical ductal hyperplasia (ADH) (100% for PRA and 100% for PRB), but the ratio of immunopositive cases and immunohistochemical (IHC) scores was significantly smaller in ductal carcinoma in situ (DCIS) (65% for PRA and 75% for PRB) and invasive ductal carcinoma (IDC) (66% for PRA and 55% for PRB) than in PDWA and ADH. There was a significant positive correlation between IHC scores for PRA and estrogen receptor alpha (ERalpha) in IDC, DCIS and ADH but not between PRB and ERalpha. In IDC, both PRA and PRB IHC scores were significantly associated with histological grade, but there was no association between PRA or PRB status and lymph node involvement, tumor size, or prognosis of the patients. The expression of mRNAs for both PRA and PRB was detected in all six cases of IDC examined. These results suggest that both PRA and PRB are strongly associated with ERalpha in human breast and this relation may be disturbed in breast cancer.

Retinoic acid receptor and retinoid X receptor in ductal carcinoma in situ and intraductal proliferative lesions of the human breast.

Retinoic acid (RAR) and retinoid X receptors (RXR) are essential in the transcriptional actions of retinoids. To date, RAR and RXR have not been examined in precancerous lesions and / or ductal carcinoma in situ (DCIS) in human breast. Therefore, we examined RAR and RXR subtypes in DCIS (58 cases), atypical ductal hyperplasia (ADH) (32 cases), and proliferative disease without atypia (PDWA) (32 cases) to study the status of these RARs and RXRs. Immunoreactivities for RAR alpha, RXR alpha, RXR beta, and RXR gamma were all detected in the nuclei of normal ductal epithelia. Immunoreactivity for RAR beta was detected exclusively in the nuclei of myoepithelial cells, but not in normal ductal epithelia. Immunoreactivity for RAR gamma was not detected in any of the breast tissues examined except for a few cases of PDWA and ADH, and 11 cases of DCIS. The RXR alpha labeling index (LI) was significantly higher in both DCIS (mean 77.9) and ADH (mean 77.7) than in PDWA (mean 62.8) (P < 0.001). RXR beta LI was significantly lower in DCIS (mean 81.5) than in both ADH (mean 91.1) and PDWA (mean 91.9) (P = 0.0001). Immunoreactivity for RAR alpha, RXR alpha, RXR beta and RXR gamma was widely distributed compared to that of RAR beta and RAR gamma in DCIS, ADH and PDWA. RAR alpha LI was significantly correlated with Ki67 LI in DCIS (P = 0.0040), especially in estrogen receptor (ER)-positive DCIS. Our results suggest that RXRs are much more widely distributed than RARs in intraductal proliferative lesions of tne human breast, but ER-positive DCIS cases with high cell proliferative activity are associated with RAR alpha, suggesting the possible involvement of retinoids through RAR alpha in tumor cell proliferation in DCIS.

17-beta-hydroxysteroid dehydrogenase in human breast and endometrial carcinoma. A new development in intracrinology.

Intratumoral metabolism and synthesis of estrogens are considered to play very important roles in the pathogenesis and development of various sex steroid-dependent neoplasms including breast and endometrial carcinoma. 17 beta-Hydroxysteroid dehydrogenase (17 beta-HSD) isozymes catalyze the interconversion of estradiol (E(2)) and estrone (E(1)), and thereby serve to modulate the tissue levels of bioactive E(2). 17 beta-HSD type 1 primarily catalyzes the reduction of E(1) to E(2), whereas 17 beta-HSD type 2 primarily catalyzes the oxidation of E(2) to E(1). In the human breast and its disorders, 17 beta-HSD type 1 is expressed in proliferative diseases without atypia, atypical ductal hyperplasia, ductal carcinoma in situ and invasive ductal carcinoma. 17 beta-HSD type 2 is not detected in any of the lesions. In addition, 17 beta-HSD type 1 coexpression is significantly correlated with estrogen receptor status in invasive ductal carcinoma cases. These results indicate that breast carcinoma can effectively convert E(1), produced as a result of in situ aromatization, to E(2), a biologically potent estrogen, and exerts estrogenic actions on tumor cells through the estrogen receptor. On the other hand, in the human endometrium, 17 beta-HSD type 2 is expressed, but not 17 beta-HSD type 1. 17 beta-HSD type 2 is expressed in the secretory phase but not in any proliferative phase in the endometrial mucosa. The enzyme is expressed in 75% of endometrial hyperplasias and 37% of carcinoma cases. In endometrial carcinoma cases, a significant inverse correlation has been detected between 17 beta-HSD type 2 immunoreactivity and age (p < 0.02). These results indicate that oxidation of E(2) to E(1) is dominant in endometrial carcinoma, 17 beta-HSD types 1 and 2 play an important role in the regulation of in situ estrogen production in breast and endometrial carcinoma.

17 beta-Hydroxysteroid dehydrogenase type 1 and type 2 in ductal carcinoma in situ and intraductal proliferative lesions of the human breast.

17 beta-hydroxysteroid dehydrogenases (17 beta-HSDs) are involved in the interconversion of biologically active and inactive sex steroids and are considered to play important roles in the in situ metabolism of estrogen in various estrogen dependent tissues. 17 beta-HSD type 1 catalyzes primarily the reduction of estrone (E1) to estradiol (E2), whereas 17 beta-HSD type 2 catalyzes primarily the oxidation of E2 to E1. However, the possible biological roles of these estrogen metabolizing isozymes in human breast cancer, especially in carcinogenesis of the human breast, have not been examined in detail. Because of the potential roles of estrogens in the early stages of human breast carcinogenesis, we have examined the immunolocalization of 17 beta-HSD type 1 and type 2 isozymes and estrogen receptor alpha(ER alpha) in both normal human breast tissue and in breast cancers, including ductal carcinoma in situ (DCIS), proliferative disease without atypia (PDWA) or fibrocystic disease and atypical ductal hyperplasia (ADH). We also correlated these findings with clinicopathological findings, Ki67 antigen, progesterone receptor (PR), c-erbB-2, and p53. 17 beta-HSD type 2 immunoreactivity was sporadically detected in non-proliferative or Ki67 negative ductal epithelia of normal breast, but rarely in breast carcinoma cells. 17 beta-HSD type 1 immunoreactivity was detected in 12/22 (54.5%) PDWA cases, 8/26 (30.8%) ADH cases, and 25/40 (62.5%) DCIS cases, respectively. 17 beta-HSD type 1 immunoreactivity was not statistically correlated with the age of the patients, Ki67 labeling index (LI), and PR LI, p-53 and c-erbB-2 immunoreactivity. There was no significant correlation between ER alpha LI and 17 beta-HSD type 1 immunoreactivity. There was a positive correlation between ER alpha and Ki67 LI in PDWA, whereas a negative correlation was detected between ER alpha and Ki67 LI in DCIS. There was no correlation between ER alpha and Ki67 LI in ADH. These results suggest that in human breast epithelial cells, development of ADH and DCIS may be associated with the loss and/or deviation of oestrogen dependent regulation of cell proliferation.

17Beta-hydroxysteroid dehydrogenase type 1 and type 2 in human breast carcinoma: a correlation to clinicopathological parameters.

The expression of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) type 1 and type 2 was examined immunohistochemically in 111 invasive ductal carcinomas, and correlated with various clinicopathological parameters. This study investigates local regulatory mechanisms of oestrogens in human breast carcinoma. 17Beta-HSD type 1 was immunolocalized in carcinoma cells of 68 out of 111 invasive ductal carcinoma cases (61.3%). 17Beta-HSD type 2 immunoreactivity was not detected in all cases examined. A significant inverse correlation was observed between the immunohistochemical expression of 17beta-HSD type 1 and histological grade of the carcinoma (P < 0.02). There was a significant correlation between 17beta-HSD type 1 and oestrogen receptor (ER) labelling index (LI) (P < 0.05). In addition, carcinoma cells expressing immunoreactive 17beta-HSD type 1 were frequently positive for ER. 17Beta-HSD type 1 was also correlated with progesterone receptor (PR) LI (P < 0.05). There was a significant inverse correlation between 17beta-HSD type 1 and Ki-67 LI (P < 0.0001). No significant correlations were detected between 17beta-HSD type 1 and other clinicopathological parameters, including patient age, menopausal status, stage, tumour size, lymph node status and prognosis. This study suggests that 17beta-HSD type 1 plays an important role in the regulation of in situ oestradiol production in hormone-dependent breast carcinomas.

[CT scan in severe head injury with special reference to Glasgow coma scale].

CT scan demonstrates the invaluable information about the parenchymal lesions of head injuries. The parenchymal lesions were classified into 6 categories; 1) isodensity without mass effect: I(-), 2) isodensity with mass effect: I(+), 3)high density: H, 4) high-low density complex: H-L, 5) low density: L, 6) diffuse cerebral swelling: DCS. Glasgow coma scale (GCS) and outcome scale (GOS) were international practical scales for the evaluation of severity and prognosis of severe head injuries. One hundred and seventy-four cases with severe head injury were analysed. I(+), H and H-L were common findings in the group of GCS 3-6, and I(-) was in GCS 7-12 and GCS 13-15. H and H-L were not related with GCS. DCS was most common in GCS 7-12. Acute epidural hematoma was frequent in the group of GCS 13-15, and acute subdural hematoma was in GCS 3-6. The prognosis was significantly poor in the group of GCS 3-6, with the mortality of 72 percents. On the other hand, the prognosis was quite good in GCS 7-12 and GCS 13-15. There were few reports about the traumatic subarachnoid hemorrhage (SAH). SAH was one of the important risk factors in severe head injuries and it was frequently associated with I(+), H and H-L. The prognosis of the patients with SAH was most unfavorable in the presence of I(+). Number of analyses were reported about the traumatic intraventricular hemorrhage (IVH). IVH was also one of the powerful risk factors and this was commonly associated with H and H-L. The prognosis of the patients with IVH was very poor. Finally, the groups of the patients, whose prognoses turned out to be unexpected results from GCS on admission, were analysed. First, the age was an important factor. In the patients, whose prognoses were good in spite of low GCS, I(-) was a mostly common finding, while SAH, IVH and obscured cisterns, esp. basal and quadrigeminal, were less common. In the patients, whose prognoses were poor despite of favorable GCS, H and H-L were common findings. SAH and IVH were also common. The poor prognosis was induced by secondary systemic complications, such as pneumonia and meningitis, etc.

Effect of detergents on sterol synthesis in a cell-free system of yeast.

In order to obtain information about the reactivity of enzymes in sterol synthesis of yeast, the effects of some detergents were investigated. Among the detergents used, Triton X-100 was found to exert a unique action, and its effect on the incorporation of 14C-labeled acetate, mevalonate, farnesyl pyrophosphate, or S-adenosyl-L-methionine into squalene, 2,3-oxidosqualene, and sterols in a cell-free system was examined. Triton X-100 showed virtually no effect on the enzyme activities in the reactions from acetyl CoA to farnesyl pyrophosphate, but it had a marked effect on reactions from farnesyl pyrophosphate to ergosterol. Evidence was obtained suggesting that Triton X-100 apparently activated squalene synthetase (EC 2.5.1.21) but inhibited squalene epoxidase (EC 1.14.99.7) and delta 24-sterol methyltransferase (EC 2.1.1.41). The activity of epoxidase was protected from the inhibition by increasing the concentration of cell-free extracts or by the prior addition of lecithin liposomes to the reaction mixture. The inhibition of methyltransferase was partially reversed by treatment with Bio-heads SM-2, but that of epoxidase was not reversed by the treatment.

Sterol-content lowering action of o-chlorobenzylchloride in yeast.

o-Chlorobenzylchloride, a simple aromatic halogen compound, was found to inhibit the growth of Saccharomyces cerevisiae and to lower the contents of sterols and fatty acids. The growth inhibition was considerably alleviated by the presence of sterols such as ergosterol and cholesterol and of unsaturated fatty acids such as oleate and linolenate. Inspection of effect of the inhibitor on the electron transport system related to the biosyntheses of these compounds revealed that the cytochrome contents and some enzyme activities in the system of the inhibited cells were much lower than those of the control cells. The features of the inhibition were similar to those of inhibition for other organisms by the hypocholesterolemic compounds such as triparanol and benzmalecene.

Prognosis of four cases of primary middle fossa arachnoid cyst in children.

The treatment and prognosis of 4 cases of primary middle fossa arachnoid cyst in children were analyzed with computed tomography, carotid angiography and Radio isotope cisternography at preoperative and postoperative states. In 2 of 4 cases the cysts reduced their sizes and in the other 2 cases they disappeared after operation. Three surgical methods were performed. The postoperative courses of all cases were good. The disappearance of cysts does not depend on the surgical methods.

Studies on delta8-delta7 isomerization and methyl transfer of sterols in ergosterol biosynthesis of yeast.

The formation of cholesta-7,24-dien-3 beta-ol and its activity as a substrate for the sterol side-chain methyltransferase in yeast have not previously been studied. Experiments with acetone-powder extracts of yeast showed that the sterol is formed from zymosterol by delta8-delta7 isomerization. However, direct conversion of cholesta-7,24-dien-3 beta-ol into zymosterol could not be demonstrated. The reversibility of the reaction was proved by the detection of 3H-incorporation into cholesta-8-en-3 beta-ol (with lathosterol as a carrier) from [3H]H2O in the medium. Incubation of cholesta-7,24-dien-3 beta-ol and S-adenosyl-L-[methyl-14C]methionine with the acetone-powder extract resulted in methylation of the sterol to form episterol. Similar incubation of zymosterol gave fecosterol and episterol, suggesting that fecosterol initially formed by the methylation was isomerized to episterol. In intact cells, however, an alternative pathway (zymosterol yields cholesta-7,24-dien-3 beta-ol yields episterol) may also operate. The relative importance of the two pathways is not known.