Yukari Nishimura

PTEN immunohistochemical expression is suppressed in G1 endometrioid adenocarcinoma of the uterine corpus

Purpose PTEN is a tumor suppressor gene that inhibits cell proliferation by regulating intracellular signaling pathways, and this activity can be abolished by mutations of the PTEN gene. This study was designed to examine the correlation of PTEN expression with the expression of cell cycle regulators and with clinicopathological parameters in endometrioid adenocarcinoma of the uterine corpus.MethodsTissue samples of 117 endometrioid adenocarcinomas in addition to those of 19 normal endometria and 20 endometrial hyperplasias were used for the study. Immunohistochemical staining for PTEN protein was performed with the labeled streptavidin-biotin method on formalin-fixed and paraffin-embedded tissue samples. PTEN expression was represented as the staining score.Results Immunohistochemistry showed that the nuclei of cells were positive for PTEN. The PTEN staining score of normal endometrium was significantly higher in the proliferative phase than in the secretory phase. The scores of various endometrial hyperplasias were not significantly different from each other, regardless of the type of hyperplasia. The PTEN staining scores of endometrioid adenocarcinomas were 7.6±5.2 in G1, 9.6±5.2 in G2, and 11.9±3.7 in G3, and increased significantly as the histological grade increased. PTEN staining score was not significantly correlated with clinicopathological parameters such as FIGO stage, myometrial invasion, lymph-vascular space invasion (LVSI), lymph node metastasis or group, but was significantly correlated with labeling indices (LIs) of cell cycle regulators such as Ki-67, cdk2, cyclin A, cyclin D1, cyclin E, p27, and p53. The PTEN staining score of p53-wild cases was significantly lower than that of p53-mutant ones, but there was no significant difference of the score in cases with different PTEN gene status. PTEN expression was significantly lower in cases with both high levels of estrogen receptor and progesterone receptor.Conclusion PTEN protein expression was decreased in well-differentiated and less growth-aggressive endometrial carcinoma with wild-type p53 gene and high levels of ER and PR. This suggests that disturbed PTEN expression occurs in an early phase of the tumorigenesis of well-differentiated endometrial carcinoma.

High expression of skp2 correlates with poor prognosis in endometrial endometrioid adenocarcinoma

Purpose: Skp2 interacts with the degradation of cyclin-dependent kinase inhibitor p27. This study aimed to investigate the correlation of skp2 expression with the expression of p27 and other cell cycle regulators, and clinicopathological parameters in endometrial endometrioid adenocarcinoma. Methods: Tissue samples of 136 endometrioid adenocarcinomas, in addition to 20 endometrial hyperplasias and 20 normal endometria, were immunohistochemically stained for skp2. The expression was represented as a labeling index (LI), which indicates the percentage of positive nuclei. Results: Skp2 staining was localized in the nuclei of the glandular cells of the proliferative phase endometrium, and endometrial hyperplasia and carcinoma cells. Skp2 expression was increased significantly in those of higher histological grade. The high level of skp2 expression was significantly correlated with the presence of lymph node metastasis and lymph-vascular space involvement. The LI of skp2 in endometrial carcinoma was significantly correlated with that of p27, Ki-67, cdk2, cyclin A, cyclin D1, cyclin E, p53 and PTEN. The high level of skp2 expression (LI≧20%) was significantly correlated with the patients’ poor survival. Conclusions: The skp2 level might have increased due to p27 accumulation and may be a good indicator of proliferative activity and poor prognosis.

Immunohistochemical expression of cyclin E in endometrial adenocarcinoma (endometrioid type) and its clinicopathological significance

PurposeCyclin E is known as a G1-S phase regulatory protein and its abnormal expression has been implicated in cellular proliferation. This study aimed to investigate the correlation of cyclin E expression with tumorigenesis of the endometrium, proliferative activity, and clinicopathological features of endometrial adenocarcinoma.MethodsImmunohistochemical staining for cyclin E in addition to cyclin-dependent kinase 2 (cdk2), Ki67, p27, and p53 was performed by the labeled streptavidin-biotin method on formalin-fixed, paraffin-embedded tissues of normal endometria (20 cases), endometrial hyperplasias (20 cases), and endometrial adenocarcinomas (endometrioid type) (127 cases). Positive staining was expressed as a labeling index (LI) based on percentages of positive nuclei in tumor cells.ResultsImmunohistochemistry showed that the nuclei of the cells were positive for cyclin E. Both proliferative and secretory endometria, and endometrial hyperplasia regardless of type were negligible for cyclin E expression. The expression in normal endometrium and hyperplasia was significantly less than that in endometrial adenocarcinomas (P<0.0001). LIs of cyclin E in well-differentiated, moderately differentiated, and poorly differentiated endometrial adenocarcinomas were 31.5±33.3%, 37.8±31.9%, and 51.1±30.8%, respectively. Cyclin E expression increased significantly more in histological grades. The LI of cyclin E in carcinoma was positively correlated with that of cdk2, Ki67, and p53 but not with p27. The cyclin E expression was correlated with myometrial invasion and lymph-vascular space involvement, but not with FIGO stage, lymph node metastasis, coexisting endometrial hyperplasia, estrogen receptor, progesterone receptor, and menopause.ConclusionCyclin E as a complex with cdk2 is associated with carcinogenesis and disease progression in endometrial adenocarcinoma, and might be a prognostic indicator of endometrial adenocarcinoma.

Cytologic scoring of endometrioid adenocarcinoma of the endometrium.

Endometrial carcinoma is one of the most frequent malignancies in the female genital tract, and its incidence has been increasing in Japan. Histologic grade is an important factor for organizing treatment strategies, including hormone therapy, and for predicting the prognosis of the patient. The objective of this study was to evaluate the applicability and usefulness of cytologic scoring in assessing the morphologic differentiation of endometrioid adenocarcinomas of the endometrium using endometrial smears.

Liquid-based preparation for endometrial cytology—Usefulness for predicting the prognosis of endometrial carcinoma preoperatively

The authors evaluated the applicability and usefulness of immunocytochemical staining for cyclin A, p53, estrogen receptor α (ER‐α), and progesterone receptor B (PR‐B) as a preoperative prognostic indicators for endometrial carcinoma using endometrial cytology with the liquid‐based cytology (LBC) method.

Expression of tumor suppressor gene product p14ARF in endometrioid adenocarcinoma of the uterine corpus.

Summary:p14ARF activates p53 by inhibiting MDM2 expression and arrests the cell cycle in G1 and G2/M. Abnormal p14ARF expression has been reported in various human cancers. This study investigated p14ARF expression in endometrioid adenocarcinoma of the uterine corpus in an attempt to clarify its correlation with other cell cycle-regulators and clinicopathologic parameters. The specimen studied consisted of 124 endometrioid adenocarcinomas, 20 normal endometria, and 20 endometrial hyperplasias. Immunohistochemical staining of formalin-fixed and paraffin-embedded tissues was performed using a Catalyzed Signal Amplification System. Cells with >5% positive staining were classified as positive for p14ARF. A staining score of 1 was adopted when the percentage of positive nuclei was <5%, a score of 2 when it was 5 to 50%, and a score of 3 when it was >50%. In normal endometrium, the frequency of positive staining in the proliferative phase and secretory phase was 50% (4/8) and 58.3% (7/12), with staining scores of 1.8±0.9 and 1.6±0.5, respectively. The frequency of staining in simple hyperplasia (SH), complex hyperplasia (CH), and complex atypical hyperplasia (CAH) was 88.9% (8/9), 25% (1/4), and 42.9% (3/7), respectively; the staining scores were 1.9±0.3, 1.3±0.5, and 1.4±0.5, respectively. Among endometrioid adenocarcinomas, the frequency of staining of well-differentiated (G1), moderately differentiated (G2), and poorly differentiated (G3) adenocarcinomas was 69% (49/71), 64% (16/25), and 42.9% (12/28) respectively, with staining scores of 2.1±0.8, 2±0.9, and 1.8±1, respectively. Thus expression levels of p14ARF were higher in G1 tumors than in normal endometria or endometrial hyperplasias, and the frequency of its staining in endometrioid carcinomas was inversely correlated with histologic grade. The staining score for endometrioid adenocarcinomas also was inversely correlated with the labeling index (LI) of Ki-67, but not with that of cyclin A, cyclin D1, cyclin E, cdk2, p27, p53, or other clinicopathologic parameters. In conclusion, p14ARF expression correlated with histologic grade and Ki-67, but not other prognostic factors in endometrioid adenocarcinoma. Long-term follow-up studies are needed to analyze the significance of p14ARF expression in these tumors.

Cytological significance of abnormal squamous cells in urinary cytology.

The aim of this study was to evaluate the significance of abnormal squamous cells (ASCs) in urinary cytology to clarify whether finding of ASCs could improve diagnostic accuracy. A total of 3,812 urine specimens were reviewed. We focused on three parameters of ASCs, necrotic debris, and ASC clusters, and linked them to histological diagnosis and clinical information. ASCs were identified in 34 (0.9%) specimens from 21 different patients. The incidence of ASCs was higher in females than in males. The 34 urine specimens were categorized as voided urine (16 cases), bladder‐catheterized urine (17 cases), and bladder‐washed fluid (1 case). Six (28.6%) of 21 patients were histologically diagnosed as having combined urothelial carcinoma and squamous cell carcinoma (SCC). Eight patients (38.1%) were histologically diagnosed as having SCC originating from sites other than the urinary tract; those urine specimens showed ASCs that were likely to have been exfoliated from malignant lesions. Necrotic debris and ASC clusters were identified in 12 specimens (35.3%) from 11 patients and 4 specimens (11.8%) from 4 patients, respectively, from a total of 34 specimens. Our results indicate that a great amount of care is needed for cytological diagnosis when attempting to recognize ASCs in urine specimens because ASCs were identified in not only SCC of the bladder but also in carcinoma or nonmalignant lesions of nonurinary tracts. Necrotic debris was found not only in patients who had malignant bladder tumors but also in those who had malignant lesions in locations other than the bladder. Diagn. Cytopathol. 2012.

Immunohistochemical, molecular, and clinicopathological analyses of urothelial carcinoma, micropapillary variant.

The prognosis of urothelial carcinoma, micropapillary variant (MPV), of the bladder has been shown to be worse than that of the conventional urothelial carcinoma (UC). However, it remains to be clarified why the MPV is more aggressive. We therefore here focused on the correlation between clinical features and histological, immunohistochemical and molecular findings for eight MPV and 35 UC, evaluating expression of MUC1, Ki‐67, p53, CD147, CD34, D2‐40, and extracellular matrix proteins. The Ki‐67 labeling index was significantly higher in UC than in MPV but densities of venous and lymphatic tumor emboli were significantly higher in the MPV cases and lymph node metastasis was more frequent, with a poorer prognosis. Tenascin‐C and fibronectin also showed significantly greater expression in MPV than in UC at the epithelial–mesenchymal interfaces. Direct sequencing showed point mutations of KRAS exon 1 in three MPV with significantly more frequency compared to UC. Occupation rate of the MPV area in the tumor showed significant inverse correlation with overall survival. Thus our histopathological findings provide clues to explaining why prognosis is poorer in the MPV than UC.

Expression of Cell Cycle Regulators in Endometrial Adenocarcinoma

Abnormal expressions of cell cycle regulators, such as cyclin-dependent kinases (cdk), cyclins, and cyclin kinase inhibitors, are supposed to play an important role in the tumorigenesis and progression of carcinoma. The aim of the present study was to investigate the correlation of the expression of these cell cycle regulators with proliferative activity and clinicopathological parameters in endometrial adenocarcinoma, and the regulation of these proteins by sex steroid hormones in endometrial cancer cells. Tissue samples of 127 endometrial adenocarcinomas (endometrioid type) were used in the present study. Immunohistochemical staining of cycle regulators was performed according to the labeled streptavidin-biotin method. Ishikawa cells, in which estrogen receptor (ER) cDNA was transfected by the SuperFect method, and HEC-265 cells positive for the progesterone receptor were used in an in vitro study. Quantitative analysis of proteins was performed by immunoblotting. Expression of cyclin D1, cyclin E, cyclin A, p53, and p27 was positively correlated with Ki-67 expression. Expression of cdk2, cyclin Dl, cyclin E, cyclin A, p53, and p27 was positively associated with histological grade. Expression of cyclin E was significantly correlated with lymphovascular space involvement (LVSI) and myometrial invasion, and expression of cyclin A was correlated with LVSI and group (coexistence with or without hyperplasia). p53 was related with stage, LVSI, myometrial invasion, and group; whereas p27 was correlated with stage, LVSI, and lymph node metastasis. Estradiol (E2) revealed a stimulatory effect on the growth of ER-transfected Ishikawa cells, in addition to enhancing their expression of cyclin A and cyclin E. In contrast, medroxyprogesterone acetate (MPA) suppressed HEC-265 cell growth after its accumulation of p27 in the cells. In conclusion, the expression of cell cycle regulators was significantly associated with cell proliferation, histological grade, and some clinicopathological parameters in endometrial adenocarcinoma. A cell line stably sensitive to E2 was established after ER cDNA was transfected into Ishikawa cells, and cyclin A and E may be involved in the enhanced cell growth induced by E2. p27 accumulation induced by MPA may be involved in the progesterone-induced growth suppression of endometrial cancer cells.

Comparative image analysis of conventional and thin-layer preparations in endometrial cytology.

We evaluated the differences in cytologic findings between conventional and thin‐layer preparations in endometrial cytology to introduce the thin‐layer method into routine cytology. Eighty patients who had undergone endometrial cytology and biopsy on the same day were selected and we compared the cytological findings between conventional‐ and thin‐layer preparations (TLP) in endometrial cytology. The numbers of neutrophils and cell clusters in the thin‐layer method were lower than those in the conventional smear (CSS) method. The average number of neutrophils in endometrioid adenocarcinoma was significantly higher than that in normal morphology endometrium and endometrial hyperplasia. Regarding the shape of the cell clusters, ball‐like patterns and round‐edged cell clusters were not identified in CSS. The average number of clusters in CSS was significantly greater than that using the TLP. The average of the nuclear area in CSS was significantly larger than that using the TLP, indicating that the nuclear areas in CSS were more uneven than that using the TLP. In the future, it is expected that liquid‐based cytology will be applied to the cytological diagnosis of a variety of lesions. The influence on cells due to fixation is considerable in liquid‐based preparations. Therefore, if we strive to pick up the differences between CSS and TLP of endometrial samples, the diagnostic accuracy of the latter could be improved. Diagn. Cytopathol. 2013.