Clinical Cancer Research Holland
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 Google Scholar
Google Scholar
Right arrow Articles by Ito, K.
Right arrow Articles by Inoue, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ito, K.
Right arrow Articles by Inoue, S.
Clinical Cancer Research Vol. 11, 7384-7391, October 15, 2005
© 2005 American Association for Cancer Research

Imaging, Diagnosis, Prognosis

14-3-3{sigma} in Endometrial Cancer–A Possible Prognostic Marker in Early-Stage Cancer

Kiyoshi Ito1, Takashi Suzuki2, Jun-ichi Akahira1,2, Michiko Sakuma1, Sumika Saitou1, Satoshi Okamoto1, Hitoshi Niikura1, Kunihiro Okamura1, Nobuo Yaegashi1, Hironobu Sasano2 and Satoshi Inoue3,4

Authors' Affiliations: Departments of 1 Obstetrics and Gynecology and 2 Pathology, Tohoku University Graduate School of Medicine, Sendai, Japan; 3 Research Center for Genomic Medicine and Department of Molecular Biology, Saitama Medical School, Saitama, Japan; and 4 Department of Geriatric Medicine, Graduate School of Medicine, University of Tokyo, Tokyo, Japan

Requests for reprints: Kiyoshi Ito, Department of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-Machi, Aoba-Ku, Sendai 980-8574, Japan. Phone: 81-22-717-7254; Fax: 81-22-717-7258; E-mail: kito{at}mail.tains.tohoku.ac.jp.


   Abstract
Top
 Abstract
Materials and Methods
 Results
 Discussion
 References
 
Purpose: We examined expression of 14-3-3{sigma}, a regulator of cell proliferation, and evaluated its clinical significance in endometrioid endometrial carcinoma.

Experimental Design: One hundred three endometrioid endometrial adenocarcinoma cases were examined using immunohistochemistry with archival specimens. We correlated this finding with various clinicopathologic variables, including the status of estrogen receptor, progesterone receptor, and MIB-1 (Ki-57).

Results: 14-3-3{sigma} Immunoreactivity was detected in 78 of 103 (75.3%) of carcinoma cases. No statistically significant correlation was detected between status of 14-3-3{sigma} and any of clinicopathologic variables examined. There was, however, a statistically significant correlation between loss of 14-3-3{sigma} expression and adverse clinical outcome of the patients (P = 0.0007). In the early stages of cancer (stages I and II), 14-3-3{sigma} immunoreactivity was absent in 5 of 10 (50.0%) patients who showed recurrence during follow-up, whereas its absence was detected in only 13 of 68 (19.1%) disease-free patients in the same period. In addition, 14-3-3{sigma} immunoreactivity was absent in 4 of 5 (80.0%) patients who died, whereas its absence was detected in only 14 of 73 (19.2%) patients who had lived during the same period. Patients whose tumors were negative for 14-3-3{sigma} were at much greater risk to develop recurrent and/or mortal disease (P = 0.0372 and 0.0067). In multivariate analysis using the Cox proportional hazards model, absence of 14-3-3{sigma} turned out to be statistically independent risk factor in disease-free survival and overall survival even in patients with early-stage disease (P = 0.0321 and 0.0191).

Conclusions: Results of our study showed that loss or absence of 14-3-3{sigma} determined by immunohistochemistry may be an important tool to identify endometrial carcinoma cases at high risk of recurrence and/or death, who are otherwise not detected by current clinical and pathologic evaluation, especially in the early stages of the disease. In addition, results of 14-3-3{sigma} immunohistochemistry in the early stage of endometrial carcinoma could contribute to planning postoperative follow-up and adjuvant therapy.

14-3-3 Proteins have been found to play important roles in the regulation of various cellular processes, such as cell cycle progression, cell growth, apoptosis, and signal transduction (1, 2). In humans, seven different 14-3-3 isoforms have been identified. 14-3-3{sigma}, a member of this family, is induced by DNA damage and is required for a stable G2 cell cycle arrest in epithelial cells. Loss of 14-3-3{sigma} expression results in malignant transformation in vitro and supports tumor formation in vivo, which suggests that this gene has tumor-suppressive properties. The 14-3-3{sigma} gene was originally identified as a p53-inducible gene responsive to DNA-damaging agents (3). In response to DNA damage, 14-3-3{sigma} is induced in a p53-dependent manner and prevents the cdc2/cyclin B1 complex from entering the nucleus. We showed previously that 14-3-3{sigma} undergoes proteolysis mediated by estrogen finger protein, which is a target of the estrogen receptor (ER) acting as an ubiquitin ligase of 14-3-3{sigma} in breast carcinoma cells (4). In addition, 14-3-3{sigma} is silenced by CpG methylation in a large proportion of human carcinomas (1, 2). The expression of 14-3-3{sigma} is shown to be frequently lost in human epithelial carcinoma, breast, gastric, lung (5–7), etc. We also reported recently that decreased expression of 14-3-3{sigma} was significantly associated with poor prognosis in epithelial ovarian cancer (8).

Endometrial carcinoma is the most common malignancy of the female genital tract, and its incidence has recently increased (9). In normal endometrium, 14-3-3{sigma} protein was expressed weakly in epithelial glandular cells (10). However, the status of 14-3-3{sigma} protein and its possible roles have never been examined in endometrial carcinoma. We reported previously the prognostic significance of p53 overexpression in endometrial cancer (11, 12). Therefore, decreased expression of 14-3-3{sigma} may possibly have an important role in the development of endometrial cancer, because 14-3-3{sigma} is directly regulated by p53.

To study the possible correlation between status of 14-3-3{sigma} protein and prognosis of the patients, we examined its immunoreactivity in 103 cases of endometrioid endometrial cancer and correlated the findings with clinical outcome of the patients.


   Materials and Methods
Top
 Abstract
 Materials and Methods
Results
 Discussion
 References
 
Patients and tissues. Twenty-five normal cycling human endometria (15 proliferative phase and 10 secretory phase) and 103 endometrioid endometrial adenocarcinoma (49 well differentiated, 32 moderately differentiated, and 22 poorly differentiated; 66 stage I, 12 stage II, 22 stage III, and 3 stage IV) were retrieved from surgical pathology files of Tohoku University Hospital (Sendai, Japan). The protocol for this study was approved by the Ethics Committee at Tohoku University Graduate School of Medicine (Sendai, Japan). All carcinoma specimens were obtained from surgery. We obtained nonpathologic endometria from hysterectomy specimens performed due to carcinoma in situ of the uterine cervix at Tohoku University Hospital. All endometrial carcinoma specimens were obtained from hysterectomy. Median follow-up time of the patients examined in this study was 60 months (range, 2-148 months). Disease-free survival and overall survival were calculated from the time of initial surgery to recurrence and/or death or the date of last contact. Survival times of patients still alive or lost to follow-up were censored in December 2004. Clinicopathologic findings of these patients, including age, histology, stage, grade, and preoperative therapy, were retrieved by review of patient charts. A standard primary treatment for endometrial carcinoma at Tohoku University Hospital was surgery consisting of total abdominal hysterectomy, salpingo-oopholectomy, pelvic and/or para-aortic lymphadenectomy, and peritoneal washing cytology. A total of 85 of 103 (83%) patients underwent complete surgery. Six of 85 patients had lymph node metastasis. The remaining 18 (17%) patients underwent total abdominal hysterectomy and salpingo-oopholectomy without lymphadenectomy because of obesity and/or poor performance status. None of these patients had received preoperative chemotherapy and/or hormonal therapy or pelvic irradiation. No patient had used oral contraceptives. The lesions were classified according to the Histological Typing of Female Genital Tract Tumors by the WHO and staged according to the International Federation of Gynecology and Obstetrics system (13, 14). Sixty-eight of 103 patients received pelvic radiation therapy (50 Gy) or three to six courses of chemotherapy consisting of the cisplatin-based combination regimen CAP (60-70 mg/m2 cisplatin, 40 mg/m2 doxorubicin, and 500 mg/body cyclophosphamide) after operation. Patients who had early-stage and low-grade disease (stage Ia, grade 1; stage Ia, grade 2; and stage Ib, grade 1) and patients who were associated with poor performance status did not receive any adjuvant therapy. All specimens were routinely processed (i.e., 10% formalin fixed for 24-48 hours), paraffin embedded, and thin sectioned (3 µm).

Immunohistochemistry. Immunohistochemical analysis was done employing the streptavidin-biotin amplification method using a Histofine kit (Nichirei, Tokyo, Japan) as described previously in detail by the authors (15). Polyclonal antibody for 14-3-3{sigma} (N-14) was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). The characteristics of the primary antibodies employed in this study are summarized in Table 1. For immunostaining of 14-3-3{sigma}, p53, ER{alpha}, progesterone receptor (PR), and Ki-67, the slides were heated in an autoclave at 121°C for 5 minutes in citric acid buffer [2 mmol/L citric acid and 9 mmol/L trisodium citrate dehydrate (pH 6.0)] following deparaffinization for antigen retrieval. The dilutions of the primary antibodies used for our studies were as follows: 14-3-3{sigma}, 1:100; p53, 1:40; ER{alpha}, 1:2; PR, 1:30; and Ki-67, 1:50. 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 nonneoplastic breast epithelial tissue were used as positive controls for 14-3-3{sigma}, and breast cancer was also used as positive control for ER{alpha}. As a negative control, normal rabbit or mouse IgG was used instead of primary antibodies.


View this table:
[in this window]
[in a new window]

  		Table 1. Summary of primary antibodies used in this study

 
Semiquantitative analysis of immunohistochemical staining. For evaluation of ER{alpha}, PR, and Ki-67 immunoreactivity, labeling index was obtained in glandular or carcinoma cells as described by Utsunomiya et al. (16) with some modifications. In cases immunopositive for ER{alpha}, PR, and Ki-67, >1,000 glandular or carcinoma cells were counted in each case by two of the authors (K.I. and T.S.) independently after reviewing the slides and determining the areas of evaluation simultaneously with a double-headed microscope. The percentage of immunoreactivity (i.e., labeling index) was subsequently determined. Cases with interobserver differences of >5%, which occurred in 3% to 7% of the cases examined, were reevaluated together by two of the authors above using double-headed light microscopy. Intraobserver differences were <5% when examining the same selected fields of representative cases. The mean value was obtained in cases with interobserver differences of <5%. As immunoreactivities of 14-3-3{sigma} and p53 were relatively homogeneous and clearly distinguishable as positive or negative, carcinoma cells were classified into the two groups without much differently (+, carcinoma cells with positive immunoreactivity; –, carcinoma cells with no immunoreactivity) by two of the same authors above.

Statistical analyses. Statistical analysis was done using SAS software version 5.0 (StatView, Cary, NC). The statistical difference between 14-3-3{sigma} and characteristics of the patients was evaluated in a cross-table using the {chi}2 test. Correlation between 14-3-3{sigma} and p53, ER{alpha}, PR, and Ki-67 immunoreactivity was also assessed using Mann-Whitney U test. Overall and disease-free survival curves were generated according to the Kaplan-Meier method, and the statistical significance was calculated using a log-rank test. Univariate and multivariate analyses were evaluated with Cox proportional hazards model. A result was considered significant when the P < 0.05.


   Results
Top
 Abstract
 Materials and Methods
 Results
Discussion
 References
 
Normal cycling endometrium. 14-3-3{sigma} Immunoreactivity was detected in the cytoplasm of glandular cells but not in the stromal cells of all the cases examined. Marked 14-3-3{sigma} immunoreactivity was detected in the glandular cells of secretory phase mucosa compared with those of proliferative phase mucosa (Fig. 1A and B).



View larger version (116K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 1. Immunohistochemistry of 14-3-3{sigma} expression: A, proliferative phase; B, secretory phase; C and D, endometrioid endometrial adenocarcinoma (G1); E and F, endometrioid endometrial adenocarcinoma (G3). 14-3-3{sigma} Immunoreactivity was detected in the cytoplasm of glandular cells. 14-3-3{sigma} Immunoreactivity was also detected in the cytoplasm of carcinoma cells. C and E, positive cases; D and F, negative cases. Original magnification, x400.

 
Association of 14-3-3{sigma} expression with clinicopathologic variables and estrogen receptor {alpha}, progesterone receptor, Ki-67, and p53 immunoreactivity in patients with endometrial cancer. 14-3-3{sigma} Immunoreactivity was detected in the cytoplasm of epithelial cancer cells, although ER{alpha}, PR, Ki-67, and p53 were confined exclusively to the nuclei of epithelial cells (Fig. 1C-F). 14-3-3{sigma} Immunoreactivity was present in 78 of 103 (75.3%) cases of endometrioid endometrial carcinoma. The correlation between 14-3-3{sigma} immunoreactivity and clinicopathologic variables, including ER{alpha}, PR, Ki-67, and p53 immunoreactivity, was examined. As seen in Table 2, no statistically significant correlation was detected between status of 14-3-3{sigma} immunoreactivity and any of the variables examined in this study. The status of 14-3-3{sigma} immunoreactivity tended to be inversely correlated with that of p53, but the correlation did not reach statistical significance. There were no statistically significant correlations between status of lymph node metastasis and 14-3-3{sigma} expression (P = 0.1456).


View this table:
[in this window]
[in a new window]

  		Table 2. Correlation between 14-3-3{sigma} immunoreactivity and clinicopathologic variables in endometrial cancer

 
Association of 14-3-3{sigma} expression with disease-free survival and overall survival in patients with endometrial cancer. 14-3-3{sigma} Immunoreactivity was evaluated as a prognostic variable in the 103 cases using univariate analysis (Table 3; Cox proportional hazards model). In 103 cases, 14-3-3{sigma} immunoreactivity was absent in 7 of 16 patients (43.8%) who showed recurrence during follow-up, whereas loss of its immunoreactivity was detected only in 18 of 87 (20.7%) disease-free patients for the same clinical follow-up period. 14-3-3{sigma} Immunoreactivity was also absent in 6 of 9 (66.7%) patients who died, whereas loss of its immunoreactivity was detected only in 19 of 94 (20.2%) patients who had lived during the same period. Patients whose tumors were associated with absence of 14-3-3{sigma} expression were at much greater risk to develop recurrent and/or mortal disease (P = 0.0382 and 0.0041). Indicators of clinical outcome of the patients, including ER, PR, and p53 status and histologic grade, were likewise significantly associated with poor outcome. Patients whose tumors were negative for 14-3-3{sigma} had also significantly worse disease-free survival and overall survival rates than 14-3-3{sigma}-positive ones using log-rank tests (Fig. 2A and B; P = 0.0298 and 0.0007).


View this table:
[in this window]
[in a new window]

  		Table 3. Univariate analyses of predictors of disease-free survival and overall survival for 103 patients with endometrial cancer

 


View larger version (13K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 2. A, correlation between 14-3-3{sigma} immunoreactivity and recurrence for patients with endometrial cancer. B, correlation between 14-3-3{sigma} immunoreactivity and survival for patients with endometrial cancer.

 
To determine whether the prognostic value of 14-3-3{sigma} expression was independent of other risk factors associated with clinical outcome, we examined the data using multivariate analysis. The prognostic factors examined were 14-3-3{sigma}, ER, PR, and p53 status, stage, and histologic grade. The findings are summarized in Table 4. Absence of 14-3-3{sigma} expression was independently statistically significant as risk factor in disease-free survival and overall survival of the patients (P = 0.0297 and 0.0148). PR status was an independent risk factor only in disease-free survival, and histologic grade was an independent risk factor only in overall survival. ER status turned out not to be independent prognostic indicator in both disease-free survival and overall survival. Disease-free survival and overall survival were not significantly different between the two groups who received radiation therapy or chemotherapy (data not shown).


View this table:
[in this window]
[in a new window]

  		Table 4. Multivariate analyses of predictors of disease-free survival and overall survival for 103 patients with endometrial cancer

 
Significance of 14-3-3{sigma} status in patients with early-stage disease of endometrial cancer. 14-3-3{sigma} Expression was then evaluated as a prognostic variable in 78 cases with early-stage disease (stage I and II) using univariate analysis (Table 5; Cox proportional hazards model). In the total of 78 cases, 14-3-3{sigma} immunoreactivity was absent in 5 of 10 (50.0%) patients who showed recurrence during follow-up, whereas loss of its immunoreactivity was detected only in 13 of 68 (19.1%) disease-free patients for the same period. 14-3-3{sigma} Immunoreactivity was also not detected in 4 of 5 (80.0%) patients who died, whereas absence of its immunoreactivity was detected only in 14 of 73 (19.2%) patients who had lived during the same period. Patients whose tumors did not show 14-3-3{sigma} immunoreactivity were at much greater risk to develop recurrent and/or mortal disease (P = 0.0372 and 0.0067). Patients whose tumors were negative for 14-3-3{sigma} also had significantly worse disease-free survival and overall survival rates than 14-3-3{sigma}-positive ones using log-rank tests (Fig. 3A and B; P = 0.0251 and 0.0002). In advanced-stage disease (stage III and IV), there was a trend for 14-3-3{sigma}-negative cases to undergo aggressive biological behavior than 14-3-3{sigma}-positive ones, although the differences did not reach statistical significance (data not shown). Multivariate analysis was done and summarized in Table 6. Absence of 14-3-3{sigma} immunoreactivity was independently statistically significant as risk factor in disease-free survival and overall survival in patients with early-stage disease of endometrial carcinoma (P = 0.0317 and 0.0229).


View this table:
[in this window]
[in a new window]

  		Table 5. Univariate analyses of predictors of disease-free survival and overall survival for 78 patients with stage I and II endometrial cancer

 


View larger version (12K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 3. A, correlation between 14-3-3{sigma} immunoreactivity and recurrence for patients with early-stage (stage I and II) endometrial cancer. B, correlation between 14-3-3{sigma} immunoreactivity and survival for patients with early-stage (stage I and II) endometrial cancer.

 

View this table:
[in this window]
[in a new window]

  		Table 6. Multivariate analyses of predictors of disease-free survival and overall survival for 78 patients with stage I and II endometrial cancer

 
Therefore, we examined the subgroup with completely surgically staged node-negative (International Federation of Gynecology and Obstetrics stage I and II) endometrial adenocarcinoma (Table 7). Absence of 14-3-3{sigma} immunoreactivity still turned out to be independently statistically significant as risk factor in disease-free survival (P = 0.0245) although not significant in overall survival (P = 0.0646) of the patients.


View this table:
[in this window]
[in a new window]

  		Table 7. Multivariate analyses of predictors of disease-free survival and overall survival for 64 patients with completely surgically staged node-negative (FIGO stage I and II) endometrial cancer

 

   Discussion
Top
 Abstract
 Materials and Methods
 Results
 Discussion
References
 
This is the first study that examined the status of 14-3-3{sigma} protein and its possible roles in conjunction with clinical outcome of the patients in endometrial carcinoma. The 14-3-3{sigma} gene is well-known to be induced after DNA damage in a p53-dependent manner and to play an important role in the G2 checkpoint by sequestering the cdc2/cyclin B1 complex (1, 2). An inactivation of 14-3-3{sigma} is also currently considered to play an important role in tumor development and/or progression. However, it is also true that 14-3-3{sigma} may play a different role in tumor development and/or progression among different human organs. In urinary bladder carcinoma, for example, 14-3-3{sigma} is highly up-regulated in pure squamous cell carcinoma, whereas it is down-regulated in invasive bladder urothelial cell carcinoma (17). In breast carcinoma, loss of 14-3-3{sigma} expression becomes marked in the progression from atypical hyperplastic lesions to ductal carcinoma in situ (18, 19). Loss of 14-3-3{sigma} protein was also reported in prostate carcinoma and its precursors (20–22). Therefore, loss or absence of 14-3-3{sigma} expression is generally considered an early event during carcinogenesis in both breast and prostate carcinoma (18–22). Ostergaard et al. (23) reported that less differentiated bladder squamous cell carcinoma was associated with decreased expression of 14-3-3{sigma}. We showed recently that loss of 14-3-3{sigma} expression was correlated with advanced disease and/or high-grade tumor and significantly associated with poor prognosis in epithelial ovarian carcinoma (8). In our present study, the frequency of absence of 14-3-3{sigma} immunoreactivity in clinically early disease and/or low-grade tumor was similar with that in advanced-stage and/or high-grade tumor, although decreased status of 14-3-3{sigma} immunoreactivity was significantly associated with poor prognosis in endometrioid endometrial cancer. These results suggest that the loss of 14-3-3{sigma} expression in endometrioid endometrial cancer may be associated with an aggressive biological characteristics, which play an important role in prognosis and/or recurrence, although it could be a relatively early event during their carcinogenesis.

In our present study, there were no significant differences of the findings between cases of early-stage and advanced-stage cancer, although advanced-stage cancer cases tended to be associated with worse prognosis than early-stage cases. These findings are considered to be due to the following reasons: the relatively small number of advanced-stage cancer cases, especially only 3 stage IV cases, and the fact that 15 of 22 (70%) cases of stage III examined were stage IIIa. Cases of stage IIIa, especially cytologic stage IIIa (positive peritoneal cytology alone), has been shown to be associated with much better prognosis than those of stage IIIc (24, 25). However, it awaits further investigation for clarifying the possible role of decreased status of 14-3-3{sigma} immunoreactivity in advanced-stage endometrial carcinoma cases.

Endometrial carcinoma is the most common pelvic gynecologic carcinoma, and 80% to 90% of all cases are in clinically early stage (26). Five-year survival data of the patients revealed ~10% to 20% mortality in early-stage disease (26). There have been many controversies on the possible use of adjuvant therapy in patients of early-stage endometrial carcinoma (27–31). Results of large randomized trial (the Post-Operative Radiation Therapy in Endometrial Carcinoma) showed no significant differences between survivals of the patients with or without adjuvant therapy in stage I endometrial adenocarcinoma. However, analysis of their study was limited because complete surgical staging was not a requirement for entry of the patients into the protocol (29). Very recently, Keys et al. showed no significant differences between survival of the patients with or without adjuvant therapy in completely surgically staged node-negative intermediate risk (International Federation of Gynecology and Obstetrics stage Ib, Ic, II occult) endometrial adenocarcinoma (Gynecologic Oncology Group study). The estimated 4-year survival was 86% in the group with no additional therapy arm and 92% for whole radiation therapy arm, with no statistical difference between these two groups (31). Therefore, identification of additional prognostic markers could provide the information to avoid unnecessary adjuvant therapy and to plan effective systemic treatment. Several studies have attempted to identify prognostic factors of the patients with early-stage endometrial cancer. However, none of them have provided satisfactory results. Fiumicino et al. (32) showed that microsatellite instability was an independent indicator of recurrence in early-stage endometrial adenocarcinoma, but Maxwell et al. (33) reported that microsatellite instability is rather a favorable prognostic factor. In addition, MacDonald et al. (34) and Basil et al. (35) both independently reported the lack of any correlation between microsatellite instability and clinical outcome in endometrial cancer. Recently, Powell et al. (36) examined the prognostic significance of rDNA methylation and showed that tumor rDNA level turned out to be significant prognostic factor for both disease-free survival and overall survival in early-stage endometrial cancer. Powell et al. therefore identified the prognostic indicator of early-stage endometrial carcinoma. However, their methods require sufficient quantity of frozen specimens, and patients with small tumors often did not have adequate tumor tissue for examination in clinical early stage of endometrial carcinoma, which may limit the clinical value of this interesting prognostic marker.

In our study, we studied archival or surgical pathology materials and analyzed a remarkable number of the cases with follow-up data to show possible correlation between absence of 14-3-3{sigma} and adverse clinical outcome using immunohistochemistry, which is a simple and useful method in surgical pathology specimens. In early-stage endometrial cancer, 14-3-3{sigma} immunoreactivity was not detected in 5 of 10 (50.0%) patients who had recurrence during clinical follow-up, whereas absence of its immunoreactivity was detected only in 13 of 68 (19.1%) disease-free patients during the same period. 14-3-3{sigma} Immunoreactivity was not detected in 4 of 5 (80.0%) patients who died, whereas loss of its immunoreactivity was detected only in 14 of 73 (19.2%) patients who had lived for the same period of clinical follow-up. Absence of 14-3-3{sigma} expression was independently statistically significant as risk factor in disease-free survival and overall survival in patients in the early stage of the disease. Additionally, even in the subgroup with completely surgically staged node-negative (International Federation of Gynecology and Obstetrics surgical stage I and II) endometrial adenocarcinoma, absence of 14-3-3{sigma} immunoreactivity still turned out to be independently statistically significant as risk factor in disease-free survival although not significant in overall survival of the patients.

These findings all indicate that absence of 14-3-3{sigma} protein determined by immunohistochemistry could be a very important tool to identify the patients at high risk of recurrence and/or death, who are otherwise not detected by current clinical and pathologic evaluation, especially in the early stage of endometrial carcinoma. In addition, results of 14-3-3{sigma} immunohistochemistry in early stage of endometrial carcinoma could contribute to planning postoperative follow-up and adjuvant therapy.


   Footnotes
 
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.

Received 1/26/05; revised 6/23/05; accepted 7/14/05.


   References
Top
 Abstract
 Materials and Methods
 Results
 Discussion
 References
 

  1. Hermeking H. The 14-3-3 cancer connection. Nat Rev Cancer 2003;3:931–43.[CrossRef][Medline]
  2. Wilker E, Yaffe MB. 14-3-3 Proteins—a focus on cancer and human disease. J Mol Cell Cardiol 2004;37:633–42.[CrossRef][Medline]
  3. Hermeking H, Lengauer C, Polyak K, et al. 14-3-3{sigma} is a p53-regulated inhibitor of G2-M progression. Mol Cell 1997;1:3–11.[CrossRef][Medline]
  4. Urano T, Saito T, Tsukui T, et al. Efp targets 14-3-3{sigma} for proteolysis and promotes breast tumour growth. Nature 2002;417:871–5.[CrossRef][Medline]
  5. Ferguson AT, Evron E, Umbricht CB, et al. High frequency of hypermethylation at the 14-3-3{sigma} locus leads to gene silencing in breast cancer. Proc Natl Acad Sci U S A 2000;97:6049–54.[Abstract/Free Full Text]
  6. Suzuki H, Itoh F, Toyota M, Kikuchi T, Kakiuchi H, Imai K. Inactivation of the 14-3-3{sigma} gene is associated with 5' CpG island hypermethylation in human cancers. Cancer Res 2000;60:4353–7.[Abstract/Free Full Text]
  7. Osada H, Tatematsu Y, Yatabe Y, et al. Frequent and histological type-specific inactivation of 14-3-3{sigma} in human lung cancers. Oncogene 2002;21:2418–24.[CrossRef][Medline]
  8. Akahira J, Sugihashi Y, Suzuki T, et al. Decreased expression of 14-3-3{sigma} is associated with advanced disease in human epithelial ovarian cancer: its correlation with aberrant DNA methylation. Clin Cancer Res 2004;10:2687–93.[Abstract/Free Full Text]
  9. Parker SL, Tong T, Bolden S, Wingo PA. Cancer statistics. CA Cancer J Clin 1996;46:5–27.[Abstract]
  10. Nakajima T, Shimooka H, Weixa P, et al. Immunohistochemical demonstration of 14-3-3{sigma} protein in normal human tissues and lung cancers, and the preponderance of its strong expression in epithelial cells of squamous cell lineage. Pathol Int 2003;53:353–60.[CrossRef][Medline]
  11. Ito K, Sasano H, Matsunaga G, et al. Correlations between p21 expression and clinicopathological findings, p53 gene and protein alterations, and survival in patients with endometrial carcinoma. J Pathol 1997;183:318–24.[CrossRef][Medline]
  12. Ito K, Watanabe K, Nasim S, et al. Prognostic significance of p53 overexpression in endometrial cancer. Cancer Res 1994;54:4667–70.[Abstract/Free Full Text]
  13. Tavassoli FA, Devilee P. Pathology and genetics of tumours of the breast and female genital organs. In: WHO classification of tumours. WHO, Lyon; 2003. p. 113–45.
  14. Creasman WT. Announcement FIGO stages: 1988 revisions. Gynecol Oncol 1989;35:125–7.[CrossRef]
  15. Ito K, Suzuki T, Moriya T, et al. Retinoid receptors in the human endometrium and its disorders: a possible modulator of 17ß-hydroxysteroid dehydrogenase. J Clin Endocrinol Metab 2001;86:2721–7.[Abstract/Free Full Text]
  16. Utsunomiya H, Suzuki T, Kaneko C, et al. The analyses of 17ß-hydroxysteroid dehydrogenase isozymes in human endometrial hyperplasia and carcinoma. J Clin Endocrinol Metab 2001;86:3436–43.[Abstract/Free Full Text]
  17. Moreira JM, Gromov P, Celis JE. Expression of the tumor suppressor protein 14-3-3{sigma} is down-regulated in invasive transitional cell carcinomas of the urinary bladder undergoing epithelial-to-mesenchymal transition. Mol Cell Proteomics 2004;3:410–9.[Abstract/Free Full Text]
  18. Simooka H, Oyama T, Sano T, Horiguchi J, Nakajima T. Immunohistochemical analysis of 14-3-3{sigma} and related proteins in hyperplastic and neoplastic breast lesions, with particular reference to early carcinogenesis. Pathol Int 2004;54:595–602.[CrossRef][Medline]
  19. Umbricht CB, Evron E, Gabrielson E, Ferguson A, Marks J, Sukumar S. Hypermethylation of 14-3-3{sigma} (stratifin) is an early event in breast cancer. Oncogene 2001;20:3348–53.[CrossRef][Medline]
  20. Cheng L, Pan CX, Zhang JT, et al. Loss of 14-3-3{sigma} in prostate cancer and its precursors. Clin Cancer Res 2004;10:3064–8.[Abstract/Free Full Text]
  21. Urano T, Takahashi S, Suzuki T, et al. 14-3-3{sigma} is down-regulated in human prostate cancer. Biochem Biophys Res Commun 2004;319:795–800.[CrossRef][Medline]
  22. Lodygin D, Diebold J, Hermeking H. Prostate cancer is characterized by epigenetic silencing of 14-3-3{sigma} expression. Oncogene 2004;23:9034–41.[CrossRef][Medline]
  23. Ostergaard M, Rasmussen HH, Nielsen HV, et al. Proteome profiling of bladder squamous cell carcinomas: identification of markers that define their degree of differentiation. Cancer Res 1997;57:4111–7.[Abstract/Free Full Text]
  24. Kasamatsu T, Onda T, Katsumata N, et al. Prognostic significance of positive peritoneal cytology in endometrial carcinoma confined to the uterus. Br J Cancer 2003;88:245–50.[Medline]
  25. Tebeu PM, Popowski Y, Verkooijen HM, et al. Positive peritoneal cytology in early-stage endometrial cancer does not influence prognosis. Br J Cancer 2004;91:720–4.[Medline]
  26. Creasman WT, Odicino F, Maisonneuve P, et al. Carcinoma of the corpus uteri. Int J Gynaecol Obstet 2003;83 Suppl 1:79–118.
  27. Ball HG. Do we know the best therapy for early endometrial cancer? Gynecol Oncol 1996;60:173–5.[Medline]
  28. Lanciano RM, Greven KM. Adjuvant treatment for endometrial cancer: who needs it? Gynecol Oncol 1995;57:135–7.[Medline]
  29. Creutzberg CL, van Putten WL, Koper PC, et al. Surgery and postoperative radiotherapy versus surgery alone for patients with stage-1 endometrial carcinoma: multicentre randomised trial. PORTEC Study Group. Post Operative Radiation Therapy in Endometrial Carcinoma. Lancet 2000;355:1404–11.[CrossRef][Medline]
  30. Orr JW, Jr., Roland PY, Leichter D, Orr PF. Endometrial cancer: is surgical staging necessary? Curr Opin Oncol 2001;13:408–12.[Medline]
  31. Keys HM, Roberts JA, Brunetto VL, et al. A phase III trial of surgery with or without adjunctive external pelvic radiation therapy in intermediate risk endometrial adenocarcinoma: a Gynecologic Oncology Group study. Gynecol Oncol 2004;92:744–51.[CrossRef][Medline]
  32. Fiumicino S, Ercoli A, Ferrandina G, et al. Microsatellite instability is an independent indicator of recurrence in sporadic stage I-II endometrial adenocarcinoma. J Clin Oncol 2001;19:1008–14.[Abstract/Free Full Text]
  33. Maxwell GL, Risinger JI, Alvarez AA, Barrett JC, Berchuck A. Favorable survival associated with microsatellite instability in endometrioid endometrial cancers. Obstet Gynecol 2001;97:417–22.[Abstract/Free Full Text]
  34. MacDonald ND, Salvesen HB, Ryan A, Iversen OE, Akslen LA, Jacobs IJ. Frequency and prognostic impact of microsatellite instability in a large population-based study of endometrial carcinomas. Cancer Res 2000;60:1750–52.[Abstract/Free Full Text]
  35. Basil JB, Goodfellow PJ, Rader JS, Mutch DG, Herzog TJ. Clinical significance of microsatellite instability in endometrial carcinoma. Cancer 2000;89:1758–64.[CrossRef][Medline]
  36. Powell MA, Mutch DG, Rader JS, Herzog TJ, Huang TH, Goodfellow PJ. Ribosomal DNA methylation in patients with endometrial carcinoma: an independent prognostic marker. Cancer 2002;94:2941–52.[CrossRef][Medline]




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 Google Scholar
Google Scholar
Right arrow Articles by Ito, K.
Right arrow Articles by Inoue, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ito, K.
Right arrow Articles by Inoue, S.

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