Clinical Cancer Research Infection and Cancer: Biology, Therapeutics, and Prevention
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Utsunomiya, H.
Right arrow Articles by Sasano, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Utsunomiya, H.
Right arrow Articles by Sasano, H.
Clinical Cancer Research Vol. 10, 5850-5856, September 1, 2004
© 2004 American Association for Cancer Research

Molecular Oncology, Markers, Clinical Correlates

Steroid Sulfatase and Estrogen Sulfotransferase in Human Endometrial Carcinoma

Hiroki Utsunomiya1, Kiyoshi Ito1, Takashi Suzuki2, Takako Kitamura1, Chika Kaneko2, Taisuke Nakata3, Hitoshi Niikura1, Kunihiro Okamura1, Nobuo Yaegashi1 and Hironobu Sasano2

1 Departments of Obstetrics and Gynecology and 2 Pathology, Tohoku University School of Medicine, Sendai; and 3 Kyowa Hakko Kogyo Co., Ltd., Tokyo, Japan


   ABSTRACT
Top
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Purpose: Intratumoral metabolism and synthesis of estrogens are considered to play important roles in the pathogenesis and/or development of human endometrial carcinoma. Steroid sulfatase hydrolyzes biologically inactive estrogen sulfates to active estrogens, whereas estrogen sulfotransferase sulfonates estrogens to estrogen sulfates. However, the status of steroid sulfatase and/or estrogen sulfotransferase in human endometrial carcinoma has not been examined.

Experimental Design: We first examined the expression of steroid sulfatase and estrogen sulfotransferase in 6 normal endometrium and 76 endometrial carcinoma using immunohistochemistry to elucidate the possible involvement of steroid sulfatase and estrogen sulfotransferase. We then evaluated the enzymatic activity and the semiquantitative analysis of mRNA using reverse transcription-PCR in 21 endometrial carcinomas. We correlated these findings with various clinicopathological parameters including the expression of aromatase, 17ß-hydroxysteroid dehydrogenase type 1 and type 2.

Results: Steroid sulfatase and estrogen sulfotransferase immunoreactivity was detected in 65 of 76 (86%) and 22 of 76 (29%) cases, respectively. Results of immunoreactivity for steroid sulfatase and estrogen sulfotransferase were significantly correlated with those of enzymatic activity and semiquantitative analysis of mRNA. No significant correlations were detected among the expression of the enzymes involved in intratumoral estrogen metabolism. There was a significant correlation between steroid sulfatase/estrogen sulfotransferase ratio and clinical outcomes of the patients. However, there were no significant differences between steroid sulfatase or estrogen sulfotransferase and estrogen receptor, progesterone receptor, Ki67, histologic grade, or clinical outcomes of the patients.

Conclusions: Results of our study demonstrated that increased steroid sulfatase and decreased estrogen sulfotransferase expression in human endometrial carcinomas may result in increased availability of biologically active estrogens and may be related to estrogen-dependent biological features of carcinoma.


   INTRODUCTION
Top
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Endometrial carcinoma is one of the most common malignancies in developed countries, and its incidence has increased recently (1) . In situ estrogen metabolism, including its synthesis and degradation, has been considered to play a very important role in the development and/or progression of various human estrogen-dependent neoplasms including endometrial carcinoma (2) . In endometrial carcinoma, in situ 17ß-estradiol availability has been demonstrated to be closely related to the pathogenesis and development of endometrial proliferative disorders including endometrial hyperplasia and carcinoma, especially of the endometrioid type (3) . Aromatase catalyzes circulating androgens, which are androstenedione and testosterone, into estrone (E1) and 17ß-estradiol, respectively (4 , 5) . The enzyme 17ß-hydroxysteroid dehydrogenase catalyzes the reversible intercon-version of E1 and 17ß-estradiol. 17ß-Hydroxysteroid dehydrogenase type 1 catalyzes the 17ß-reduction of biologically weak E1 to strong 17ß-estradiol (6, 7, 8) , whereas 17ß-hydroxysteroid dehydrogenase type 2 preferentially catalyzes the oxidation of 17ß-estradiol to E1 (9) .

A major circulating form of plasma estrogen is estrogen sulfate (E1S and E2S), a biologically inactive form of estrogen. E1S and E2S have a relatively long half-life in the peripheral blood (10) , where serum levels of E1S and E2S are known to be 10-fold higher than those of unconjugated E1 or 17ß-estradiol (11) . It was reported recently that in situ estrogen activity in breast cancer, which is estrogen dependent as well as endometrial cancer, may be mainly regulated by the status of intratumoral steroid sulfatase (12 , 13) . Steroid sulfatase hydrolyzes biologically inactive estrogen sulfates to active estrogens. Estrogen sulfotransferase (SULT 1E1 or STE gene) is a member of the superfamily of steroid sulfotransferases and sulfonates estrogens to estrogen sulfates (14, 15, 16) . Therefore, it is suggested that estrogen sulfotransferase, especially the balance between the levels of intratumoral steroid sulfatase and estrogen sulfotransferase, may also play an important role in the regulation of in situ estrogen levels in human endometrial carcinoma. Steroid sulfatase and estrogen sulfotransferase activities have been examined in estrogen-dependent neoplasms such as endometrial and breast cancers (17) . However, to date, steroid sulfatase and estrogen sulfotransferase mRNA and protein expressions have not been examined in human endometrial carcinoma. In addition, the comparison among the enzymes involved in intratumoral estrogen production and metabolism has not been reported in human endometrial carcinoma. Therefore, in this study we first examined the expression of steroid sulfatase and estrogen sulfotransferase in 6 normal cycling endometrium and 76 endometrial carcinomas using immunohistochemistry to elucidate the possible involvement of steroid sulfatase and estrogen sulfotransferase. We then studied the enzymatic activity and semiquantitative analysis of mRNA by reverse transcription-PCR (RT-PCR) for steroid sulfatase and estrogen sulfotransferase in endometrial carcinomas. We subsequently correlated these findings with the results of Aromatase, 17ß-hydroxysteroid dehydrogenase type 1, type 2, and clinicopathological parameters to study the biological and/or clinical significance of steroid sulfatase and estrogen sulfotransferase.


   MATERIALS AND METHODS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Tissue Preparation.
Six normal cycling human endometria (3 proliferative phase and 3 secretory phase) and 76 endometrial endometrioid adenocarcinomas (33 well differentiated, 26 moderately differentiated, and 17 poorly differentiated; 44 stage I, 14 stage II, 17 stage III, and 1 stage IV) were obtained from surgical pathology files of Tohoku University Hospital. This study was approved by the Ethical Committee of Tohoku University School of Medicine. We obtained nonpathological endometria from hysterectomy specimens performed due to carcinoma in situ of the uterine cervix. Endometrial carcinoma specimens were obtained from hysterectomy. All of the patients examined had not received irradiation or chemotherapy before surgery. The histopathological classification in each specimen was evaluated according to Fédération Internationale des Gynaecologistes et Obstetristes histologic grading system for endometrial carcinoma in 1988 (18) .

The specimens were all routinely processed (i.e.,10% formalin fixed for 24 to 48 h), paraffin embedded, and thin sectioned (3 µm).

Antibodies.
Rabbit polyclonal antibody for estrogen sulfotransferase (PV-P2237) was purchased from Medical Biological Laboratory (Nagoya, Japan). This antibody was raised against the synthetic NH2-terminal peptide of human estrogen sulfotransferase corresponding to amino acids 1–13. The affinity-purified monoclonal antibody for steroid sulfatase (KM1049) was raised against the enzyme purified from human placenta, which recognized the steroid sulfatase peptide corresponding to amino acids 420–428. 17ß-Hydroxysteroid dehydrogenase type 1 antibody (polyclonal) and 17ß-hydroxysteroid dehydrogenase type 2 antibody (monoclonal) were kindly provided by Dr. Matti Poutanen at the University of Oulu (Oulu, Finland) and Dr. Stefan Andersson at the University of Texas Southwestern Medical Center (Dallas, TX), respectively. Aromatase antibody (polyclonal) was provided by Dr. Nobuhiro Harada at the Fujita Health University School of Medicine (Aichi, Japan). Monoclonal antibodies for estrogen receptor (ER1D5), progesterone receptor (MAB429), and Ki67 (MIB1) were purchased from Immunotech (Marseille, France), Chemicon (Temecula, CA), and DAKO (Carpinteria, CA), respectively. Utilization of these antibodies for immunohistochemistry has been reported previously (19) .

Immunohistochemistry.
Immunohistochemical analyses were performed using the streptavidin-biotin amplification method using EnVision (DAKO Co. Ltd., Carpinteria, CA) for steroid sulfatase and using a Histofine kit (Nichirei, Tokyo, Japan) for estrogen sulfotransferase, Aromatase, estrogen receptor, progesterone receptor, Ki67, 17ß-hydroxysteroid dehydrogenase type 1, and type 2. The dilutions of the primary antibodies used in this study were as follows: 1:1,500 estrogen sulfotransferase, 1:9,000 steroid sulfatase, 1:800 17ß-hydroxysteroid dehydrogenase type 1, 1:5 17ß-hydroxysteroid dehydrogenase type 2, 1:750 Aromatase, 1:2 estrogen receptor, 1:30 progesterone receptor, and 1:50 Ki67.

The antigen-antibody complex was visualized with 3.3'-diaminobenzidine solution [1 mmol/L 3.3'-diaminobenzidine, 50 mmol/L Tris-HCl buffer (pH 7.6), and 0.006% H2O2], and counterstained with hematoxylin. Tissue sections of full-term placenta were used as positive control for steroid sulfatase, and normal liver was also used as positive control for estrogen sulfotransferase. As for negative controls, normal rabbit or mouse IgG was used instead of the primary antibodies. No specific immunoreactivity was detected in these tissue sections.

Scoring of Immunoreactivity.
For evaluation of steroid sulfatase and estrogen sulfotransferase immunoreactivity, we determined the labeling index (LI: i.e., the percentage of positive cells) according to the report by Sasano et al. (20) . As in previous studies, two of the authors (H. U. and T. S.) independently divided the cases into the following two groups: +, > 5% positive cells and –: <5% positive-cell immunoreactivity (20 , 21) . Scoring of 17ß-hydroxysteroid dehydrogenase type 1, type 2, estrogen receptor, progesterone receptor, and Ki67 in gland or carcinoma cells was reported previously (21) .

Enzyme Assay.
Twenty one carcinoma cases of fresh-frozen tissues (i.e., the cases immediately frozen in liquid nitrogen and stored at –80°C) were available for examination of enzymatic assay. Estrogen sulfotransferase was assayed as described previously (15) . The samples were homogenized at 4°C in phosphate buffer [100 mmol/L KCl, 10 mmol/L KH2PO4, 10 mmol/L Na2HPO4, and 1 mmol/L EDTA (pH 7.5)], and centrifuged for 15 minutes at 1000 x g. The upper layer was used as the enzyme source. Approximately 0.2 mg of protein were added in each assay, and the reaction contained 50 mmol/L Tris-HCl (pH 7.4) and 7 mmol/L MgCl2, and E1 contained [3H]E1 at 20 nM. Reactions were started with the addition of 3'-phosphoadenosine 5'-phosphosulfate to final concentration of 20 µmol/L in a final volume of 0.125 ml. The reaction mixtures were incubated at 37°C for 30 minutes, and the reactions were terminated with the addition of 4.0 mL of chloroform followed by the addition of 0.375 mL 0.25 mol/L Tris-HCl (pH 8.7) to alkalinize the solution. The reaction mixtures were centrifuged at 600 x g for 5 minutes to separate E1S (aqueous phase) from E1 (organic phase). Synthesis of the tritiated E1S was determined with a liquid scintillation counter (Beckman, LC-6500). The steroid sulfatase activity was assayed according to Utaaker and Stoa (22) with slight modifications.

Briefly, enzyme solution (~0.2 mg protein) was mixed with E1S containing [6,7-3H]E1S (1.6 x 105 dpm, 0.5 pmol/L) at 20 µmol/L and added to a reaction volume up to 15 mL with PBS (-) containing 25 mmol/L sucrose and 4 mmol/L Nicotinamide. The reaction mixture was incubated at 37°C for 60 minutes in a shaking water bath. The enzyme reaction was terminated with the addition of toluene and mixed by vortex mixer for 1 minute. The reaction mixtures were centrifuged at 600 x g for 5 minutes to separate E1S (aqueous phase) from E1 (organic phase). The toluene layer was collected, and [3H] radioactivity was measured by liquid scintillation counter (LC-6500, Beckman), which is equivalent to E1 formed. Incubation condition of these assays was designed so that the formation of product was linear.

Semiquantitative Analysis of mRNA.
Twenty one specimen of fresh-frozen tissues (i.e., the cases immediately frozen in liquid nitrogen and stored at –80°C) were available for examination. Total RNA was extracted by homogenizing frozen tissue samples in 1 mL of TRIzol reagent (Life Technologies, Inc., Grand Island, NY) followed by a phenol-chloroform phase extraction and isopropanol precipitation. All of the RNA samples were quantified by spectrophotometry and stored at –80°C until they were processed for reverse transcription. The SUPERSCRIPT Preamplification system reverse transcription kit (Life Technologies, Inc.) was used in the synthesis and amplification of cDNA. cDNA was synthesized from total RNA (2 µg) using 25 ng/µL oligo(dT)12–18 primer (Life Technologies, Inc., Gaithersburg, MD) on a PTC-200 Peltier Thermal Cycler DNA Engine (MJ Research, Inc., Watertown, MA). To test for the presence of genomic DNA contamination, we performed the reverse transcription step in the absence of SUPERSCRIPT II RNase H– Reverse Transcriptase (Life Technologies, Inc.,) followed by PCR. RT-PCR products lacking reverse transcriptase in the initial reverse transcription step were run on an ethidium bromide-stained 2% agarose gel. No band was observed in these samples (data not shown). The resulting cDNA was used as a template for real-time PCR. Real-time PCR was carried out with the Light Cycler System (Roche Diagnostics GmbH, Mannheim, Germany) using the DNA binding dye SYBER Green I (Roche Diagnostics GmbH) for the detection of PCR products. The primer sequences used in this study are as follows: estrogen sulfotransferase [NM005420; FWD 5'-AGAGGAGCTTGTGGACAGGA-3' and REV 5'-GGCGACAATTTCTGGTTCAT-3'] (14) , steroid sulfatase [M16505; FWD 5'-AGGGTCTGGGTGTGTCTGTC-3' and REV 5'-ACTGCAACGCCTACTTAAATG-3'] (23) , and glyceraldehyde-3-phosphate dehydrogenase [M33197; FWD 5'-TGAACGGGAAGCTCACTGG-3' and REV 5'-TCCACCACCCTGTTGCTGTA-3'] (24) . PCR was set up using 2 mmol/L MgCl2, 10 pmol/L of each primer, and 2.5 units of TaqDNA polymerase (Life Technologies, Inc.). An initial denaturing step of 95°C for 1 minute was followed by 40 cycles, respectively, of 95°C for 0 seconds; 15 seconds annealing at 60°C (steroid sulfatase and glyceraldehyde-3-phosphate dehydrogenase) and 58°C (estrogen sulfotransferase); and extension for 15 seconds at 72°C. The fluorescence intensity of the double strand-specific SYBER Green I, which reflects the amount of formed specific PCR products, was read by the LightCycler at 85°C after the end of each extension step. After PCR, these products were resolved on a 2% agarose ethidium bromide gel. Images were captured with Polaroid film under UV transillumination. In initial experiments, PCR products were purified and subjected to direct sequencing (ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction kit and ABI PRISM 310 Genetic Analyzer, Perkin-Elmer Corp., PE Applied Biosystems, Foster City, CA) to verify amplification of the correct sequences. As a positive control, frozen tissues of placenta were used for steroid sulfatase (25) , and liver (HuH7 human hepatocellular carcinoma cells) was used for estrogen sulfotransferase (16) . Negative control experiments lacked cDNA substrate to check for the presence of exogenous contamination DNA. No amplified products were observed under these conditions. The mRNA levels of steroid sulfatase and estrogen sulfotransferase in each case are summarized as a ratio of glyceraldehyde-3-phosphate dehydrogenase and evaluated as a ratio (%) compared with that of each positive control. Although conventional quantitative PCR requires the utilization of a purified plasma cDNA in the construction of a standard curve, we found that we were able to semiquantify our PCR products with the Light Cycler using purified cDNA of known concentrations. Other studies to date have used a similar protocol to semiquantify PCR products with the LightCycler (26 , 27) .

Statistical Analysis.
Values for patient age and LIs of estrogen receptor, progesterone receptor, and Ki67 were presented as mean ±95% confidence interval. Association between steroid sulfatase/estrogen sulfotransferase and these parameters was evaluated using Welch’s t test. We also studied the statistical differences between patients for steroid sulfatase/estrogen sulfotransferase and histologic grade in a cross-table using the {chi}2 test. Overall and disease-free survival curves were generated according to the Kaplan-Meier method. P values < 0.05 were considered as significant.


   RESULTS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Normal Cycling Endometrium.
Estrogen sulfotransferase immunoreactive protein was detected only in the cytoplasm of glandular cells in the secretory phase (Fig. 1A)Citation . Estrogen sulfotransferase immunoreactivity was not detected in either epithelium or stromal cells of proliferative phase endometrium (Fig. 1B)Citation . Steroid sulfatase immunoreactivity was not detected in any of the cases examined.



View larger version (134K):
[in this window]
[in a new window]
  		Fig. 1. Immunohistochemistry for estrogen sulfotransferase in normal endometrium obtained during secretory phase (A) and proliferative phase (B). Original magnification, x200.

 
Endometrial Carcinoma.
Steroid sulfatase and estrogen sulfotransferase immunoreactivity was detected in the cytoplasm of carcinoma cells (Fig. 2, A and B)Citation but not in stromal cells. Steroid sulfatase and estrogen sulfotransferase immunoreactivity was detected in 65 of 76 (86%) and 22 of 76 (29%) cases, respectively. There were no significant correlations between steroid sulfatase or estrogen sulfotransferase and estrogen receptor LI, progesterone receptor LI, Ki67 LI, histologic grade, or clinical outcomes of the patients (Tables 1Citation and 2Citation ). However, there was a significant positive correlation between steroid sulfatase/estrogen sulfotransferase ratio and clinical outcomes of the patients [P < 0.05; alive (3.93 ± 2.95) versus dead (8.35 ± 4.53)]. There wereno associations among the expression of steroid sulfatase, estrogen sulfotransferase, Aromatase, 17ß-hydroxysteroid dehydrogenase type 1, and type 2 (Table 3)Citation .



View larger version (118K):
[in this window]
[in a new window]
  		Fig. 2. Immunohistochemistry for steroid sulfatase (A) and estrogen sulfotransferase (B) in endometrial endometrioid adenocarcinoma. Original magnification, x200.

 

View this table:
[in this window]
[in a new window]
  		Table 1 Correlation between STS immunoreactivity and other parameters in human endometrial carcinoma

 

View this table:
[in this window]
[in a new window]
  		Table 2 Correlation between EST immunoreactivity and other parameters in human endometrial carcinoma

 

View this table:
[in this window]
[in a new window]
  		Table 3 Correlation between immunoreactivity of the enzymes

 
For enzyme activity, the results of all of the cases are summarized in Table 4Citation . There was a statistically significant positive correlation between immunoreactivity and enzyme activity for steroid sulfatase [P < 0.01; + (0.48 ± 0.22 nmol/mg/hour) versus – (0.18 ± 0.10 nmol/mg/hour)] and estrogen sulfotransferase [P < 0.05; + (0.41 ± 0.46 nmol/mg/hour) versus – (0.09 ± 0.03 nmol/mg/hour)].


View this table:
[in this window]
[in a new window]
  		Table 4 Twenty one cases for enzyme assay and RT-PCR

 
For RT-PCR, the results of all cases are summarized in Table 4Citation . There was a statistically significant positive correlation between steroid sulfatase immunoreactivity and results of steroid sulfatase semiquantitative analysis of mRNA (RT-PCR) [P < 0.05; + (42.2 ± 22.3) versus – (20.2 ± 9.3)]. There was also significant positive correlation between estrogen sulfotransferase immunoreactivity and results of estrogen sulfotransferase RT-PCR analysis [P < 0.05; + (35.0 ± 28.1) versus – (15.9 ± 7.4)].


   DISCUSSION
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
Estrogens, especially 17ß-estradiol, have been demonstrated to contribute greatly to the development and progression of the great majority of endometrial carcinoma (28 , 29) . In addition, in situ estrogen metabolism, including its synthesis and degradation, has been considered recently to play a very important role in the development and progression of various human estrogen-dependent neoplasms including endometrial carcinoma especially in the postmenopausal subjects (2) .

Two principal pathways are implicated in the last steps of 17ß-estradiol formation in humans. One is through the aromatization of androstenedione or testosterone to E1 or 17ß-estradiol, which was considered a major pathway for peripheral estrogen production (4 , 5) . However, conversion of E1S and E2S into E1 and 17ß-estradiol has been also postulated as a source of peripheral estrogen production (30 , 31) . For instance, the concentrations of E1S and E2S in breast cancer tissues have been reported to be significantly higher than the circulating plasma levels (32) . In addition, quantitative determinations of estrogens in breast cancer tissues demonstrated that the "estrogen sulfatase pathway" is 40–500 times higher than that of the "aromatase pathway" (32 , 33) . Therefore, peripheral steroid sulfatase/estrogen sulfotransferase is considered to play very important roles in estrogen metabolism and actions in human breast cancer.

Steroid sulfatase is known as a single enzyme that hydrolyzes several sulfated steroids, including not only E1S and E2S but also dehydroepiandrosterone sulfate and cholesterol sulfate (34 , 35) . Recent studies demonstrated that the bioactive androgen 5{alpha}-dihydrotestosterone is locally produced from dehydroepiandrosterone (36 , 37) in human breast cancer tissues. A great majority of estrogen receptor-positive breast carcinomas are also positive for androgen receptor (38) . In vitro studies also demonstrated that 5{alpha}-dihydrotestosterone decreases estrogen-induced cell proliferation in human breast cancer cells (39) . Therefore, steroid sulfatase may possibly be involved in the local production of both androgens and estrogens in endometrial carcinoma as well as breast carcinomas. Additional investigations are required to clarify the biological significance of these interesting findings.

Estrogen sulfotransferase expression has been reported to be induced by progesterone in the uterus (40, 41, 42) . In addition, estrogen sulfotransferase activities were reported to be highest during the secretory phase of the menstrual cycle in the porcine uterus, which suggests a possible correlation between uterine estrogen sulfotransferase activity and plasma progesterone levels (43) . However, we reported previously that 17ß-hydroxysteroid dehydrogenase type 2 was also present in the endometrial glandular epithelium of secretory phase, but 17ß-hydroxysteroid dehydrogenase type 1 was not detected in any of the phases (21) . Estrogen sulfotransferase in endometrial carcinoma was markedly diminished compared with normal endometrium of secretory phase, which is in the same manner as pattern of 17ß-hydroxysteroid dehydrogenase type 2 expression in human endometrium (21) . These results suggest that the expression of estrogen sulfotransferase and 17ß-hydroxysteroid dehydrogenase type 2, both of which decrease in situ estrogen activity, may be regulated by progesterone.

It was reported that steroid sulfatase activities in endometrial cancer tissues were significantly higher than those in normal endometrial tissues (17) . In addition, estrogen sulfotransferase activities were reported to be highest during the secretory phase of the menstrual cycle (40 , 41) . In our present study, steroid sulfatase was absent through the menstrual cycle, and estrogen sulfotransferase was highly expressed in normal endometrium of secretory phase, which were consistent with the results of previous studies. On the other hand, steroid sulfatase was strongly expressed in 86% of the endometrial carcinoma cases, but estrogen sulfotransferase was expressed only in 29% of the cases. Decreased expression of estrogen sulfotransferase and 17ß-hydroxysteroid dehydrogenase type 2 in endometrial carcinoma may be related to abundant in situ availability of 17ß-estradiol. Therefore, in situ estrogen activity in endometrial carcinoma is much higher than that in normal endometrium. However, there are no correlations between steroid sulfatase or estrogen sulfotransferase and clinicopathological parameters and clinical outcomes of the patients. Therefore, these data indicated that not only steroid sulfatase and estrogen sulfotransferase but also Aromatase and 17ß-hydroxysteroid dehydrogenase type 2 may independently contribute to the regulation of in situ estrogen availability and/or activity in human endometrial carcinoma. In addition, there was a significant positive correlation between steroid sulfatase/estrogen sulfotransferase ratio and clinical outcomes of the patients. Therefore, the results of the present study demonstrated that increased steroid sulfatase and decreased estrogen sulfotransferase expression in human endometrial carcinomas result in increased in situ availability of biologically active estrogens and may be related to various estrogen-dependent features of carcinoma.

In endometrial carcinoma, 17ß-hydroxysteroid dehydrogenase type 1 expression was not detected (21) . This is in contrast to the study of 17ß-hydroxysteroid dehydrogenase type 1 in breast cancer in which nearly half of the cases demonstrated 17ß-hydroxysteroid dehydrogenase type 1 immunoreactivity in carcinoma cells, and 17ß-hydroxysteroid dehydrogenase type 2 was not expressed (20 , 44) . We have reported recently that estrogen sulfotransferase is an independent prognostic factor in human breast carcinoma (45) . However, there is no correlation between estrogen sulfotransferase and clinical outcomes of the patients in this study. These results indicate that intratumoral estrogen metabolism is different between human breast and endometrial carcinoma, although both of them are sex steroid-dependent malignancies.

In conclusion, the results from our study also suggest that induction of estrogen sulfotransferase and 17ß-hydroxysteroid dehydrogenase type 2 and/or inhibition of steroid sulfatase and aromatase may also have possible important therapeutic potential as an endocrine therapy for endometrial carcinoma. Utilization of selective agents designed with the intent to block the expression of enzyme related to intratumor estrogen production can result in a more specific inhibition of estrogen actions in endometrial carcinoma, which may, in turn, eventually lead to an improvement in the prognosis in some of the patients, but additional investigations are required for clarification.


   FOOTNOTES
 
Grant support: Grant-in-aid for Cancer Research 7–1 from The Ministry of Health and Welfare, Japan, a grant-in-aid for scientific research area on priority area (A-11137301) from The Ministry of Education, Science and Culture, Japan, a grant-in-aid for Scientific Research (B-11470047) from Japan Society for the Promotion of Science, and a grant from The Naitou Foundation and Suzukenn Memorial Foundation.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Requests for reprints: Hiroki Utsunomiya, Northwestern University, Reproductive Biology Research, Department of Obstetrics and Gynecology, 303 East Chicago Avenue, Tarry Building, Suite #13–756, Chicago, IL 60611. Phone: 312-503-5399; Fax: 312-503-5394; E-mail: u-kichi{at}pg8.so-net.ne.jp

Received 1/ 7/04; revised 4/ 1/04; accepted 4/ 8/04.


   REFERENCES
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. Parker SL, Tong T, Bolden S, Wingo PA. Cancer statistics. CA-Cancer J Clin, 46: 5-27, 1996.[Abstract]
  2. Silverberg SG, Mullen D, Faraci JA, Makowski EL, Miller A, Finch JL, Sutherland JV. Clinical-pathologic comparison of cases in postmenopausal women receiving exogenous estrogens. Cancer, 45: 3018-26, 1980.[CrossRef][Medline]
  3. Lippman ME, Swain SA. Endocrine-responsive cancers of humans Wilson JD Foster DW eds. . Williams textbook of endocrinology, p. 1577-97, Saunders Philadelphia 1992.
  4. Miller WR, Hawkins RA, Forrest AM. Significance of aromatase activity in human breast cancer. Cancer Res, 42: 3365-8, 1982.
  5. Miller WR, Forrest AP. Oestradiol synthesis by a human breast carcinoma. Lancet, 2: 866-8, 1974.[Medline]
  6. Peltoketo H, Isomaa V, Maentausta O, Vihko R. Complete amino acid sequence of human placental 17ß-hydroxysteroid dehydrogenase deduced from cDNA. FEBS Lett, 239: 73-7, 1988.[CrossRef][Medline]
  7. Luu-The V, Labrie C, Zhao HF, et al Characterization of cDNAs for human estradiol 17ß-dehydrogenase and assignment of the gene to chromosome 17: evidence of two mRNA species with distinct 5'-termini in human placenta. Mol Endocrinol, 3: 1301-9, 1989.[Abstract]
  8. Gast MJ, Sims HF, Murdock GL, Gast PM, Strauss AW. Isolation and sequencing of a cDNA clone encoding human placental 17ß-estradiol dehydrogenase: identification of the putative cofactor binding site. Am J Obstet Gynecol, 161: 1726-31, 1989.[Medline]
  9. Wu L, Einstein M, Geissler WM, Chan HK, Elliston KO, Andersson S. Expression cloning and characterization of human 17ß-hydroxysteroid dehydrogenase type 2, a microsomal enzyme possessing 20{alpha}-hydroxysteroid dehydrogenase activity. J Biol Chem, 268: 12964-9, 1993.[Abstract/Free Full Text]
  10. Ruder HJ, Loriaux L, Lipsett MB. Estrone sulfate: production rate and metabolism in man. J Clin Invest, 51: 1020-33, 1972.
  11. Samojlik E, Santen RJ, Worgul TJ. Plasma estrone-sulfate: assessment of reduced estrogen production during treatment of metastatic breast carcinoma. Steroids, 39: 497-507, 1982.[CrossRef][Medline]
  12. Chetrite GS, Cortes-Prieto J, Philippe JC, Wright F, Pasqualini JR. Comparison of estrogen concentrations, estrone sulfatase and aromatase activities in normal, and in cancerous, human breast tissues. J Steroid Biochem Mol Biol, 72: 23-7, 2000.[CrossRef][Medline]
  13. Pasqualini JR, Chetrite GS. Estrone sulfatase versus estrone sulfotransferase in human breast cancer: potential clinical applications. J Steroid Biochem Mol Biol, 69: 287-92, 1999.[CrossRef][Medline]
  14. Aksoy IA, Wood TC, Weinshilboum R. Human liver estrogen sulfotransferase: identification by cDNA cloning and expression. Biochem Biophys Res Commun, 200: 1621-9, 1994.[CrossRef][Medline]
  15. Falany CN, Krasnykh V, Falany JL. Bacterial expression and characterization of a cDNA for human liver estrogen sulfotransferase. J Steroid Biochem Mol Biol, 52: 529-39, 1995.[CrossRef][Medline]
  16. Dooley TP, Haldeman-Cahill R, Joiner J, Wilborn TW. Expression profiling of human sulfotransferase and sulfatase gene superfamilies in epithelial tissues and cultured cells. Biochem Biophys Res Commun, 277: 236-45, 2000.[CrossRef][Medline]
  17. Yamamoto T, Kitawaki J, Urabe M, et al Estrogen productivity of endometrium and endometrial cancer tissue; influence of aromatase on proliferation of endometrial cancer cells. J Steroid Biochem Mol Biol, 44: 463-8, 1993.[CrossRef][Medline]
  18. FIGO Stages-1988 revision. Gynecol Oncol, 35: 125-7, 1989.[CrossRef]
  19. Saeki T, Takashima S, Sasaki H, Hanai N, Salomon DS. Localization of estrone sulfatase in human breast carcinomas. Breast Cancer, 6: 331-7, 1999.[Medline]
  20. Sasano H, Frost AR, Saitoh R, et al Aromatase and 17ß-hydroxysteriod dehydrogenase type 1 in human breast carcinoma. J Clin Endocrinol Metab, 81: 4042-6, 1996.[Abstract/Free Full Text]
  21. Utsunomiya H, Suzuki T, Chika C, et al The analyses of 17ß-hydroxysteroid dehydrogenase isozymes in human endometrial hyperplasia and carcinoma. J Clin Endocrinol Metab, 86: 3436-43, 2001.[Abstract/Free Full Text]
  22. Utaaker E, Stoa KF. Oestrone sulphatase activity of the rat uterus in different hormonal states. Horm Res, 13: 180-5, 1980.[Medline]
  23. Utsumi T, Yoshimura N, Takeuchi S, et al Steroid sulfatase expression is an independent predictor of recurrence in human breast cancer. Cancer Res, 59: 377-81, 1999.[Abstract/Free Full Text]
  24. Tokunaga K, Nakamura Y, Sakata K, et al Enhanced expression of a glyceraldehyde-3-phosphate dehydrogenase gene in human lung cancers. Cancer Res, 47: 5616-9, 1987.[Abstract/Free Full Text]
  25. Salido EC, Yen PH, Barajas L, Shapiro LJ. Steroid sulfatase expression in human placenta: immunocytochemistry and in situ hybridization study. J Clin Endocrinol Metab, 70: 1564-7, 1990.[Abstract]
  26. Dumoulin FL, Nischalke HD, Leifeld L, et al Semi-quantification of human C-C chemokine mRNAs with reverse transcription/real-time PCR using multi-specific standards. J Immunol Methods, 241: 109-19, 2000.[CrossRef][Medline]
  27. Read SJ. Recovery efficiencies on nucleic acid extraction kits as measured by quantitative LightCycler PCR. Mol Pathol, 54: 86-90, 2001.[Abstract/Free Full Text]
  28. Thomas DB. Do hormones cause cancer?. Cancer, 53: 595-604, 1984.[CrossRef][Medline]
  29. Vihko R, Apter D. Endogenous steroids in the pathophysiology of breast cancer. CRC Crit. Rev Oncol/Hematol, 9: 1-16, 1989.
  30. Dao TL, Hayes C, Libby PR. Steroid sulfatase activities in human breast tumors. Proc Soc Exp Biol Med, 146: 381-4, 1974.[Medline]
  31. Pasqualini JR, Gelly C, Lecerf F. Estrogen sulfates: biological and ultrastructural responses and metabolism in MCF-7 human breast cancer cells. Breast Cancer Res. Treat, 8: 233-40, 1986.[CrossRef][Medline]
  32. Pasqualini JR, Chetrite G, Blacker C, et al Concentrations of estrone, estradiol, and estrone sulfate and evaluation of sulfatase and aromatase activities in pre- and postmenopausal breast cancer patients. J Clin Endocrinol Metab, 81: 1460-4, 1996.[Abstract]
  33. Santner SJ, Feil PD, Santen RJ. In situ estrogen production via the estrone sulfatase pathway in breast tumors: relative importance versus the aromatase pathway. J Clin Endocrinol Metab, 59: 29-33, 1984.[Abstract]
  34. Burns GR. Purification and partial characterization of arylsulphatase C from human placental microsomes. Biochim Biophys Acta, 759: 199-204, 1983.[Medline]
  35. Dibbelt L, Kuss E. Human placental steryl-sulfatase. Enzyme purification, production of antisera, and immunoblotting reactions with normal and sulfatase- deficient placentas. Biol Chem Hoppe Seyler, 367: 1223-9, 1986.[Medline]
  36. Recchione C, Venturelli E, Manzari A, Cavalleri A, Martinetti A, Secreto G. Testosterone dihydrotestosterone and oestradiol levels in postmenopausal breast cancer tissues. J Steroid Biochem Mol Biol, 52: 541-6, 1995.[CrossRef][Medline]
  37. Suzuki T, Darnel AD, Akahira JI, et al 5alpha-reductases in human breast carcinoma: possible modulator of in situ androgenic actions. J Clin Endocrinol Metab, 86: 2250-7, 2001.[Abstract/Free Full Text]
  38. Isola JJ. Immunohistochemical demonstration of androgen receptor in breast cancer and its relationship to other prognostic factors. J Pathol, 170: 31-5, 1993.[CrossRef][Medline]
  39. Poulin R, Baker D, Labrie F. Androgens inhibit basal and estrogen-induced cell proliferation in the ZR-75–1 human breast cancer cell line. Breast Cancer Res Treat, 12: 213-25, 1988.[CrossRef][Medline]
  40. Tseng L, Liu HC. Stimulation of arylsulfotransferase activity by progestins in human endometrium in vitro. J Clin Endocrinol Metab, 53: 418-21, 1981.[Abstract]
  41. Clarke CL, Adams JB, Wren BG. Induction of estrogen sulfotransferase in the human endometrium by progesterone in organ culture. J Clin Endocrinol Metab, 55: 70-5, 1982.[Abstract]
  42. Falany JL, Falany CN. Regulation of estrogen sulfotransferase in human endometrial adenocarcinoma cells by progesterone. Endocrinology, 137: 1395-401, 1996.[Abstract]
  43. Brooks SC, Rozhin J, Pack BA, et al Role of sulfate conjugation in estrogen metabolism and activity. J Toxicol Environ Health, 4: 283-300, 1978.[Medline]
  44. Poutanen M, Isomaa V, Lehto VP, Vihko R. Immunohistochemical analysis of 17ß- hydroxysteroid dehydrogenase in benign and malignant human breast tissue. Int J Cancer, 50: 386-90, 1992.[Medline]
  45. Suzuki T, Nakata T, Miki Y, et al Estrogen sulfotransferase and steroid sulfatase in human breast carcinoma. Cancer Res, 63: 2762-70, 2003.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 EndocrinologyHome page
P. A. Foster, L. W. L. Woo, B. V. L. Potter, M. J. Reed, and A. Purohit
The Use of Steroid Sulfatase Inhibitors as a Novel Therapeutic Strategy Against Hormone-Dependent Endometrial Cancer
Endocrinology, August 1, 2008; 149(8): 4035 - 4042.
[Abstract] [Full Text] [PDF]

Home page
 Hum ReprodHome page
A. Purohit, L. Fusi, J. Brosens, L.W.L. Woo, B.V.L. Potter, and M.J. Reed
Inhibition of steroid sulphatase activity in endometriotic implants by 667 COUMATE: a potential new therapy
Hum. Reprod., February 1, 2008; 23(2): 290 - 297.
[Abstract] [Full Text] [PDF]

Home page
 Hum ReprodHome page
H. Dassen, C. Punyadeera, R. Kamps, B. Delvoux, A. Van Langendonckt, J. Donnez, B. Husen, H. Thole, G. Dunselman, and P. Groothuis
Estrogen metabolizing enzymes in endometrium and endometriosis
Hum. Reprod., December 1, 2007; 22(12): 3148 - 3158.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Utsunomiya, H.
Right arrow Articles by Sasano, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Utsunomiya, H.
Right arrow Articles by Sasano, H.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online