MIB-1 (Ki-67), p53, estrogen receptor, and progesterone receptor expression in uterine smooth muscle tumors

The diagnosis of benign, uncertain malignant potential, and malignant uterine smooth muscle tumors depends on mitotic counts, nuclear atypia, and other morphologic features. This study was undertaken to evaluate the utility of selected immunohistochemical markers in differentiating these tumors. Fifteen cases of cellular leiomyoma, 7 cases of smooth muscle tumor of uncertain malignant potential (STUMP), and 12 cases of leiomyosarcoma were immunostained for MIB-1 (Ki-67), p53, estrogen receptor and progesterone receptor (PR) using monoclonal antibodies and the avidin-biotin-peroxidase method. The percentage of cells stained was subjectively assessed to the nearest 5%. One percent was used for rare positive cells. MIB-1 expression of > or =15% was seen in 11 and expression of p53 in > or =15% cells was present in 5 of 12 leiomyosarcomas. MIB-1 and/or p53 expression of >15% was seen in all 12 leiomyosarcomas but in none of the 7 STUMP or 15 cellular leiomyomas. PR was absent in 10 of 12 leiomyosarcomas but present in 7 of 7 STUMP and 14 of 15 cellular leiomyomas. MIB-1 of 5% to 10% was seen in 6 of 7 STUMP but in only 1 of 15 cellular leiomyomas. MIB-1, p53, and PR are useful in differentiating leiomyosarcoma from STUMP and cellular leiomyoma. MIB-1 is useful in distinguishing STUMP from cellular leiomyomas.     

Modulation of alpha smooth muscle actin and desmin expression in perisinusoidal cells of normal and diseased human livers.

It has been suggested that perisinusoidal liver cells (PSC) play a pivotal role in the pathogenesis of fibrocontractive changes. Using light and electron microscopic immunolocalization techniques, a series of 207 normal and pathologic human liver specimens were evaluated for the expression of alpha smooth muscle (SM) actin and desmin in this and other nonparenchymal cell types. In normal adult liver tissue, PSCs were practically devoid of desmin and exceptionally stained for alpha-SM actin, whereas this actin isoform frequently was encountered in PSCs from the embryonic to the adolescent period. A broad spectrum of pathologic conditions was accompanied by the presence of alpha-SM actin containing PSCs; these were detected preferentially in periportal or perivenular zones according to the predominant location of the underlying hepatocellular damage. The occurrence of this PSC phenotype generally was associated with fibrogenesis and was in some cases detected earlier than overt collagen accumulation. Fibrous bands subdividing liver tissue in cirrhosis and focal nodular hyperplasia, as well as desmoplastic reaction to malignant tumors, contained alpha-SM actin-rich cells admixed with variable proportions of cells coexpressing desmin. In end stages, this population was less numerous than in active fibrotic or cirrhotic processes. Using immunogold electron microscopy, alpha-SM actin was localized in microfilament bundles of typical PSCs. Our results are compatible with the assumption that the appearance of alpha-SM actin and desmin-expressing myofibroblasts results at least in part from a phenotypic modulation of PSCs.     

Comparison of the use of antibodies to alpha smooth muscle actin and desmin in pulmonary lymphangioleiomyomatosis.

To establish if antibodies to alpha smooth muscle actin (ASMA) are better than desmin as a tumour marker in pulmonary lymphangioleiomyomatosis both antisera were applied to five cases. ASMA strongly stained the muscle in all cases but desmin was negative. Five cases of cryptogenic fibrosing alveolitis showed that mature smooth muscle embedded in fibrous tissue surrounding cysts was positively stained with ASMA but the interstitial fibrous tissue was negative. ASMA is a consistent and better marker than desmin for the detection of immature smooth muscle in pulmonary lymphangioleiomyomatosis.     

Increased expression of calcium-binding protein S100 in human uterine smooth muscle tumours

S100 proteins belong to the EF-hand Ca(2+ )-binding protein family and regulate a variety of cellular processes via interaction with different target proteins. Several diseases, including cancer and melanoma, are related to the abnormal expression of S100 proteins, which are expressed in cell- and tissue-specific manners. We investigated the expression of S100 family members in human uterine smooth muscle tumours. Expression of six members of the S100 protein family: S100A1, A4, A6, A7, A10 and A11, was found in human uterine leiomyoma and myometrium tissue, but expression of other members was not detected by RT-PCR. Real-time PCR showed that S100A11 expression was significantly increased in leiomyoma compared with myometrium. Suppression of S100A11 by small interfering RNA (siRNA) led to apoptosis, and the overexpression of S100A11 inhibited apoptosis in human uterine smooth muscle tumour cells. These findings suggest that S100A11 has an anti-apoptotic function and is related to the process of growth of human uterine leiomyoma.     

Concerted upregulation of CLP36 and smooth muscle actin protein expression in human endometrium during decidualization

The human endometrium prepares for implantation of the blastocyst by reorganization of its whole cellular network. Endometrial stroma cells change their phenotype starting around the 23rd day of the menstrual cycle. These predecidual stroma cells first appear next to spiral arteries, and after implantation these cells further differentiate into decidual stroma cells. The phenotypical changes in these cells during decidualization are characterized by distinct changes in the actin filaments and filament-related proteins such as alpha-actinin. The carboxy-terminal LIM domain protein with a molecular weight of 36 kDa (CLP36) is a cytoskeletal component that has been shown to associate with contractile actin filaments and to bind to alpha-actinin supporting a role for CLP36 in cytoskeletal reorganization and signal transduction by binding to signaling proteins. The expression patterns of CLP36, alpha-actinin and actin were studied in endometrial stroma cells from different stages of the menstrual cycle and in decidual stroma cells from the 6th week of gestation until the end of pregnancy. During the menstrual cycle, CLP36 is only expressed in the luminal and glandular epithelium but not in endometrial stroma cells. During decidualization and throughout pregnancy, a parallel upregulation of CLP36 and smooth muscle actin, an early marker of decidualization in the baboon, was observed in endometrial decidual cells. Since both proteins maintain a high expression level throughout pregnancy, a role of both proteins is suggested in the stabilization of the cytoskeleton of these cells that come into close contact with invading trophoblast cells.     

S-100 protein expression by primary and metastatic adenocarcinomas

S-100 protein has been used as a marker of various lesions, including peripheral nerve sheath, cartilaginous and salivary gland tumors, chordomas, histiocytosis X, and melanomas, among others. The list of neoplasms that can express S-100 protein continues to expand. It has been suggested that staining for S-100 protein may be of aid in the differential diagnosis of amelanotic melanoma versus poorly differentiated tumors. Three hundred fifty primary and metastatic adenocarcinomas from various sites were immunostained for S-100 protein with the use of a commercially available polyclonal antibody. Forty-two percent of the adenocarcinomas tested expressed S-100 protein to varying degrees. The relative incidence of S-100-positive tumors varied with the primary sites, some expressing S-100 protein more often than others. A primary neoplasm able to express S-100 protein was usually associated with metastatic foci also expressing this marker. However, occasionally, a primary S-100-positive tumor was associated with metastasis that lacked expression of S-100. This study emphasizes the importance of testing for a panel of tumor markers in the evaluation of poorly differentiated tumors and cautions on possible difficulties that may arise in the interpretation of immunocytochemistry results.     

Thioredoxin is associated with proliferation, p53 expression and negative estrogen and progesterone receptor status in breast carcinoma

We investigated the expression of thioredoxin and thioredoxin reductase in a large set of breast invasive and in situ carcinomas by immunohistochemistry. Additionally, NF-kappa B, p53 and proliferating cell nuclear antigen (PCNA) expression was studied. Thioredoxin and thioredoxin reductase expression was located in both cytoplasmic and nuclear compartments of the cell. Cytoplasmic thioredoxin positivity was found in 67 % and nuclear in 59 % of the cases, while thioredoxin reductase was found in 55 % and 6 % of cases, respectively. Ductal carcinomas showed stronger cytoplasmic thioredoxin immunoreactivity than lobular ones. Nuclear thioredoxin positivity was more often found in in situ lesions, and lobular carcinomas were more often negative than ductal ones. Both cytoplasmic and nuclear thioredoxin-positive cases had a high proliferation measured by PCNA staining. Positive nuclear immunostaining was associated with negative estrogen and progesterone receptor status. Cases with high p53 expression showed significantly higher nuclear thioredoxin positivity, but lower thioredoxin reductase positivity. Whilst thioredoxin or thioredoxin reductase was not associated with patient survival, cases showing both cytoplasmic and nuclear thioredoxin reductase-positive tumours had a shorter disease-free interval than those with negative immunostaining.     

Immunoreactivity of alpha smooth muscle actin in salivary gland tumours: a comparison with S100 protein.

Antibodies against alpha smooth muscle actin (ASMA) and S100 protein were applied to paraffin wax embedded sections from 40 salivary gland tumours and seven normal salivary glands. The results indicate that ASMA is useful in the diagnosis of epithelial-myoepithelial carcinoma, but is otherwise only of limited use in diagnostic practice. An unexpected finding was the failure of ASMA to react with myoepitheliomas.     

The role of estrogen receptor, progesterone receptor and p53 in development of stress urinary incontinence

Recent studies have been reported the roles of the estrogen receptor (ER), progesterone receptor (PR) and p53 in the development of a pelvic organ prolapse (POP). The pathogenesis of stress urinary incontinence (SUI) is related to that of POP in the weakness of pelvic support. Therefore, this study was carried out to assess the relationship between ER, PR, p53 and the development of SUI, and to elucidate the biomolecular pathophysiology of SUI. The periurethral fascia was obtained from 6 menopausal patients diagnosed with SUI and 10 menopausal patients without SUI who visited the Department of Obstetrics and Gynecology, Severance Hospital, Seoul, Korea. The relative ER, PR and p53 protein levels in the periurethral fascia were obtained by western blot analysis and densitometry. A Mann-Whitney U test was used for statistical analysis (p < 0.05). The mean age (+/- SD) of the 16 patients was 59.0 +/- 5.5 years (range, 50-74 years). The mean body mass index was 25.2 +/- 2.7 kg/m2 (21.8 - 30.8) and the average number of vaginal deliveries was 2.8 +/- 1.9 (1.0 - 9.0). The ER level (0.33 +/- 0.17 vs. 1.86 +/- 0.83, p= 0.02) and the p53 level (1.25 +/- 0.67 vs. 4.71 +/- 2.40, p= 0.01) were lower in the experimental group. However, the PR level of the two groups were similar (0.64 +/- 0.13 vs. 0.48 +/- 0.33, p=0.56). The p53 and ER levels were significant lower in the study group. This suggests that p53 and ER might be important factors in the development of SUI. Further prospective studies about the association of ER, p53 and SUI will be needed to elucidate the molecular pathogenesis of SUI.     

Bcl-2 and p53 protein expression, apoptosis, and p53 mutation in human epithelial ovarian cancers

Bcl-2 and p53 gene products have been both linked to cell death by apoptosis. In the present study, we examined the relationship of Bcl-2 and p53 protein expression, p53 mutation and apoptosis in normal human ovaries and different types of human ovarian epithelial tumors by immunohistochemical localization, in situ terminal transferase-mediated dUTP nick end labeling and polymerase chain reaction-single strand conformation polymorphism. It was found that Bcl-2 expressed strongly in the surface epithelium of normal ovaries and benign and borderline ovarian tumors but weakly in the malignant tumors. On the contrary, strong protein expression of p53 was found in 54% (25/46) of the malignant epithelial tumors examined but similar expression of p53 was not observed in borderline and benign tumors and normal ovarian surface epithelium. A significant inverse correlation between Bcl-2 and p53 expression was found in the malignant ovarian tumors examined. p53 gene mutation at exons 5-11 was however not a pre-requisite for p53 expression in both borderline and malignant tumors. Apoptotic activities, as reflected by apoptotic indices, were low in normal ovarian surface epithelium and benign tumors but were increased in borderline and malignant tumors, with the highest average apoptotic index found in grade III malignant tumors. Statistical analyses showed a positive correlation between apoptosis and p53 expression, but similar correlation was not found between apoptosis and Bcl-2 expression. Our results also indicate that although expression of Bcl-2 is important during ovarian carcinogenesis, the Bcl-2 protein may have other roles to play apart from being a modulator of apoptosis in human ovarian epithelial cancers.     

Extent and level of fascial involvement in 100 cases with nodular fasciitis

Summary Nodular fasciitis (NF) shows a cellular proliferation which leads to widening of the fascia. Frequently unilateral or more often bilateral disruption of the fascia, with an infiltrative pattern is present. Subcutaneous fascia and surrounding fat are involved. Superficially the cellular proliferation may extend into dermal connective tissue. Deeper muscular tissue may be involved, with transitional forms or purely intramuscular changes, compatible with proliferative myositis. Proliferative myositis is considered to be a deep-seated variant of NF with muscular involvement. Intramuscular myxoma may be thought of as an intramuscular and mucoid variant of NF. A bilateral infiltrative pattern was most frequently found at all levels; in cases with muscular involvement it was always present.

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Zusammenfassung Die Beurteilung der Ausdehnung und des topographischen Niveaus der FascienverÄnderungen bei 100 FÄllen mit nodulÄrer Fasciitis ergab z.T. nur intrafasciale Zellproliferation mit spindeliger Auftreibung der Fascie. HÄufiger fand sich jedoch einseitig von der Fascie ausgehendes infiltratives Wachstum, am hÄufigsten eine vollstÄndige Unterbrechung der ursprünglichen Fascienstruktur mit beidseitiger Infiltration der Umgebung.Die nodulÄre Fasciitis befiel nicht nur die subcutane Fascie und das umgebende Fettgewebe. VerÄnderungen der oberflÄchlichen Fascie führen zu einer Mitbeteiligung der Cutis. Bei tief gelegener nodulÄrer Fasciitis greifen die VerÄnderungen auf die Muskulatur über. Es finden sich entweder übergangsformen zur proliferativen Myositis bei Befall der supramuskulÄren Fascie oder bei rein intramuskulÄren VerÄnderungen das typische Bild des proliferativen Myositis, die demnach als tiefsitzende Variante der nodulÄren Fasciitis mit Muskelbeteiligung aufgefa\t werden kann. IntramuskulÄre Myxome, die ebenfalls z.T. übergangsformen zur typischen nodulÄren Fasciitis erkennen lie\en, scheinen eine intramuskulÄre mucoide Variante der nodulÄren Fasciitis darzustellen.Insgesamt fand sich am hÄufigsten eine bilaterale Infiltration in allen Lagen der nodulÄren Fasciitis. In FÄllen mit Muskelbeteiligung fand sich ausschlie\lich eine Zellproliferation mit Zerstörungen der Fascie und bilateraler Infiltration der Umgebung.

     

Loss of heterozygosity in P53, BRCA1, and estrogen receptor genes and correlation to expression of p53 protein in ovarian epithelial tumors of different cell types and biological behavior

Loss of heterozygosity (LOH) of tumor suppressor genes (TSGs) in ovarian epithelial tumors of differing cell types and biological behavior has not been thoroughly investigated. Moreover, there have been conflicting reports correlating LOH of the p53 gene to overexpression of p53 protein. This study evaluated 34 formalin-fixed, paraffin-embedded ovarian epithelial tumors for LOH by polymerase chain reaction (PCR) for the following microsatellite markers: TP53(17p13.1/p53 gene), D17S579(17q/BRCA1 gene), and ESR (6q24-27/estrogen receptor gene). LOH of the TP53 marker was detected in 4 (44%) of 9 informative serous cystadenocarcinomas (SCa) but in 0 of 4 informative clear cell carcinomas (CCa) and 0 of 5 informative serous tumors of low malignant potential (SLMP). LOH of the BRCA1 marker was detected in 5 (83%) of 6 informative SCa, but in 1 (13%) of 8 informative CCa and 1 (14%) of 7 informative SLMP. LOH of the ESR marker was detected in 4 (50%) of 8 informative SCa, but in 0 of 4 informative CCa and 1 (16%) of 6 informative SLMP. p53 protein overexpression was present in 8 of 12 SCa but did not correlate to TP53 LOH. LOH for TP53, D17S579/ BRCA1, and ESR is common in ovarian SCa, and is observed in primary tumors as well as metastases. In contrast, these genetic alterations are less common in CCa and in the biologically less aggressive SLMP tumors. These data suggest different mechanisms of oncogenesis in ovarian epithelial tumors of different cell types and biological behavior.     

S100A6 expression in cutaneous smooth muscle neoplasms

The S100A6 protein is expressed in a variety of tissues and distinct staining patterns in S100A6 immunohistochemistry may be useful in the differential diagnosis of difficult lesions. We evaluated the staining pattern of the S100A6 antibody in 22 cases each of pilar leiomyoma (LM), angioleiomyoma (ALM), and cutaneous leiomyosarcoma (LMS). S100A6 labeled both the nucleus and cytoplasm of myocytes in positive cases. About 64% of LM and 86% ALM had positive staining to the S100A6 antibody but predominantly in a weak staining pattern. In contrast, 95% of the LMS exhibited moderate to strong staining with the S100A6 antibody. The difference in the frequency of positive cases was statistically significant in the LM vs LMS comparison (p = 0.025), but we found intensity of staining to be of greatest practical utility. Analysis between the groups taking in to consideration differences in intensity of staining using the nonparametric rank sum (Mann–Whitney U test) demonstrated that there was a statistically significant difference between LM and LMS and between ALM and LMS. Weak or absent S100A6 staining supports a diagnosis of LM, whereas strong positive staining supports a diagnosis of LMS.     

Expression of desmin and smooth muscle myosin heavy chain in dermatofibromas

BACKGROUND: The histopathologic features of dermatofibroma vary remarkably, and this diversity may occasionally cause problems in differentiating between benign and malignant mesenchymal lesions, including smooth muscle neoplasms. Immunohistochemical stains are sometimes necessary to clarify the histogenesis of a lesion. OBJECTIVE: To evaluate dermatofibromas for expression of desmin and smooth muscle myosin heavy chain (SM-MHC) antigens, which are commonly used as evidence of smooth muscle differentiation. METHODS: We studied 100 consecutive cases of dermatofibroma using hematoxylin-eosin-stained sections and immunoperoxidase staining with antibodies against desmin, SM-MHC, and smooth muscle actin. RESULTS: We found focal positivity for desmin in 9 cases, and in 2 of these cases, at least 10% of lesional cells showed strong expression. We found focal staining for SM-MHC in 10 cases, and in 2 of these cases, at least 10% of the lesional cells were positive. Regions positive for desmin and/or SM-MHC did not show definite histologic features of myogenous differentiation on hematoxylin-eosin-stained sections. All dermatofibromas expressing desmin and SM-MHC were also strongly positive for smooth muscle actin. CONCLUSIONS: About 10% of dermatofibromas show focal expression of desmin and SM-MHC, and this expression may be present in up to 10% to 15% of lesional cells. Thus, in dermal spindle cell lesions, focal expression of these muscle antigens, like that of smooth muscle actin, is not diagnostic of a smooth muscle tumor.     

Immunohistochemical expression of S-100 protein in human embryonal fat cells

The aim of the present study was to investigate immunocytochemically the presence of S-100 protein in subcutaneous fat cells during early human embryogenesis (6-12 weeks of gestation). We found that preadipocytes in the subcutaneous tissue which were at different stages of differentiation were positive for S-100 protein. The other cells in the embryonal subcutis (mesenchymal cells differentiating into fibroblasts and fibrocytes and endothelial cells) showed a negative reaction for S-100. Our results imply that the S-100 protein is expressed from the beginning of lipidogenesis and possibly acts as a factor regulating lipid storage and body fat formation. It can be used as a reliable biochemical marker of human fat cell differentiation and for distinguishing them from the mesenchymal and the fibroblast cells in the human embryonal subcutis.     

雌激素受体在大鼠血管平滑肌细胞中的表达及其调控

目的：观察血管平滑肌细胞(VSMC)中雌激素受体(ER)的表达,研究雌激素和血清(含多种生长因子)对细胞中：ER表达的影响。方法：原位杂交和RT-PCR检测大鼠主动脉中层VSMC中的ERmRNA。分别在有或无他莫昔芬(TAM)(ER拮抗剂)预处理的情况下,免疫细胞化学观察VSMC中的ER蛋白,比较雌二醇(E2)以及新生小牛血清(NCS)对VSMC中ER表达的影响。结果：VSMC中存在ERmRNA和蛋白,E2以及NCS均能上调细胞中ER的表达,TAM能阻断其效应。结论：ER是雌激素作用于VSMC的靶基因之一。     

Prognostic significance of p53 gene mutations and p53 protein expression in synovial sarcomas

Alterations to p53 seem to be of prognostic significance in soft tissue sarcomas, but their significance for synovial sarcomas has not been studied. We analysed 34 synovial sarcomas in 19 patients for p53 alterations (p53 gene mutations + p53 immunopositivity) and examined this factor for its prognostic value in a group of 15 primary tumours. DNA was prepared from paraffin-embedded tumour material by a modified proteinase K/phenol/chloroform extraction. p53 gene mutations of exons 5–8 were analysed by the PCR-SSCP-sequencing method. p53 protein expression was evaluated by immunohistochemistry using the murine monoclonal antibody DO1. We found two missense mutations (5.9%) and ten p53 immunopositive cases (29.4%). Both tumours with p53 mutations showed p53 protein expression. There was no significant correlation between p53 alteration and histological subtype, age, sex, or tumour size. The 5-year survival rate was 24.1%. Overall survival was significantly reduced in patients having synovial sarcomas with p53 alterations (P<0.001). In the multivariate Cox’s analysis, only p53 alterations (P=0.032) and tumour size (P=0.023) emerged as independent prognostic factors. We suggest that p53 alterations may be a useful prognostic indicator in synovial sarcomas, allowing rational clinical treatment and follow-up. Received: 13 April 1999 / Accepted:18 May 1999     

Relationship between p53 pathway and estrogen receptor status in endometrioid-type endometrial cancer

We analyzed the mechanism of estrogen receptor (ER) loss and status of the p53 pathway in 64 cases of endometrial cancer. 26.6% (17 of 64) of endometrial cancers lost ER. Methylation of the ER CpG island was significantly related to ER status (P = 0.0074). However, the methylation site of the ER CpG island differed between breast and endometrial cancers. The abnormal expression rate of p14ARF, MDM2, p53, and the p53 pathway were 7.8% (5 of 64), 32.8% (21 of 64), 25.0% (16 of 64) and 53.1% (34 of 64), respectively. There was no significant difference in the overexpression of MDM2 between p53-positive cases (43.8%: 7 of 16) and p53-negative cases (29.2%; 14 of 48) (P = 0.3595). Abnormal p53 was higher in grade 3 tumors (55.6%; 5 of 9) than in grade 1 and 2 tumors (20.0%; 11 of 55) (P = 0.0364). The abnormality of the p53 pathway was higher in grade 3 tumors (88.9%; 8 of 9) than in grade 1 and 2 tumors (47.3%; 26 of 55) (P = 0.0294). However, there was no significant difference in abnormal p53 pathway between ER-negative and ER-positive cases. In endometrial cancer, ER CpG island methylation was the important mechanism of ER loss. However, there was no significant relationship between the p53 pathway and ER status.