Uterine Endometrial Adenocarcinoma Research Articles

Metastatic Mullerian adenocarcinoma involving the pancreas initially misdiagnosed as an intraductal papillary mucinous neoplasm (IPMN): A case report, and lessons learned

Abstract Introduction/Objective Metastases to the pancreas from Mullerian primaries are rare and can be confused with primary pancreatic malignancies. Methods/Case Report A 49-year-old woman presented with right lower abdominal pain. She denied any prior history of malignancy but had undergone hysterectomy and bilateral salpingo-oophorectomy for unknown reasons. CT scan revealed an 8.9 cm predominantly cystic mass in the pancreas, highly suspicious of an invasive mucinous neoplasm. EUS-guided FNA of the pancreas showed large, complex papillary fronds of mucinous epithelial cells in a background of abundant extracellular mucin, findings that were interpreted as an intraductal papillary mucinous neoplasm (IPMN) with foci of high-grade dysplasia and possible adenocarcinoma. A subsequent PET/CT scan demonstrated a 3.9 cm hepatic mass and a 3.8 cm perihepatic lesion. FNA of the liver was interpreted as metastatic pancreatic adenocarcinoma. Despite aggressive treatment, her disease progressed. Molecular profiling of her liver tumor demonstrated a pathogenic KRAS G12D mutation and a PDGFRA variant of uncertain significance. Genomic (DNA) and transcriptomic (RNA) analysis indicated a 96% probability that the patient had ovarian or fallopian tube adenocarcinoma and a 4% probability of uterine endometrial adenocarcinoma rather than metastatic pancreatic adenocarcinoma. A panel of immunocytochemical stains was performed on the pancreas and liver FNAs. The tumor cells were strongly positive for CK7, CK19, ER, and WT1, focally positive for PAX8 (nuclear) and P53 (wild-type), and negative for CK20, CDX2, and SATB2. Results (if a Case Study enter NA) NA Conclusion These findings corroborated the molecular analysis suggesting that the patient had metastatic Mullerian adenocarcinoma rather than metastatic pancreatic adenocarcinoma. Her oncologic treatment was adjusted accordingly.

Pan-Cancer Analyses Reveal Oncogenic Role and Prognostic Value of F-Box Only Protein 22.

The F-box protein 22 (FBXO22), an F-box E3 ligase, has been identified to be critically involved in carcinogenesis. However, a systematic assessment of the role of FBXO22 across human cancers is lacking. Here, we performed a pan-cancer analysis to explore the role of FBXO22 in 33 cancer types using multiomic data from The Cancer Genome Atlas (TCGA). First, we found that high FBXO22 expression in multiple cancers was closely associated with poor overall survival and relapse-free survival. Next, we identified ten proteins that interact with FBXO22 and 13 of its target substrates using the STRING database and a literature search to explore the regulatory role of FBXO22 in tumorigenesis. Genes encoding these proteins were found to be significantly enriched in cell cycle negative regulation and ubiquitination pathways. This was confirmed in nonsmall cell lung cancer A549 cells, where FBXO22 overexpression enhanced cyclin-dependent kinase 4 (CDK4) protein levels and promoted cell proliferation. Similarly, overexpression or interference of FBXO22 changed the protein level of one of its substrates, PTEN. Additionally, we found that FBXO22 mutations were accompanied by altered substrate expression, especially in uterine corpus endometrial carcinoma and lung adenocarcinoma; endometrial carcinoma patients with FBXO22 genetic alterations also had better overall and relapse-free survival. Notably, FBXO22 methylation levels were also decreased in most tumors, and hypomethylation of FBXO22 was associated with poor overall survival, relapse-free interval, and progression-free interval in pancreatic adenocarcinoma. Finally, we analyzed the correlation between the abundance of tumor infiltrating lymphocytes (TILs) and FBXO22 expression, copy number variation, and methylation. Multiple algorithms revealed that high FBXO22 expression was associated with lower TIL levels, especially in lung adenocarcinoma, lung squamous cell carcinoma, and sarcoma. Taken together, our findings demonstrate that FBXO22 degrades tumor suppressor genes by ubiquitination and inhibits the cell cycle to promote nonsmall cell lung cancer progression. Our study also provides a relatively comprehensive understanding of the oncogenic role of FBXO22 in different tumors.

FAM65A as a novel prognostic biomarker in human tumors reveal by a pan-cancer analysis

BackgroundFamily with sequence similarity 65 member A (FAM65A), also known as RIPOR1, is differentially expressed between human tumor and non-tumor tissues in kinds of cancers. In addition, it was reported that the product of FAM65A may be a biomarker for cholangiocarcinoma patients. However, there is still no evidence on the relationship between the FAM65A and different types of tumors. Our study is mainly for exploring the prognostic values of FAM65A in pan-cancer and for further discovering a potential therapeutics target.MethodsWe analyzed FAM65A expression, prognostic values, genetic alteration, protein phosphorylation, immune infiltration and enrichment analysis across different types of human malignant tumors based on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. Additionally, Real-Time PCR (RT-qPCR) was performed to further confirm the roles of FAM65A in the pathogenesis of colorectal cancer.ResultsWe found that FAM65A expression was associated with the prognosis of multiple human tumors, especially colorectal cancer. Moreover, we also observed that FAM65A was highly expressed in colorectal cancer through RT-qPCR. We observed that decreasing phosphorylation level of the S351 locus in colon adenocarcinoma, uterine corpus endometrial carcinoma and lung adenocarcinoma. And the expression of FAM65A was positively related to cancer-associated fibroblasts (CAFs) infiltration in many tumors, such as colon adenocarcinoma. Therefore, FAM65A may be a potential prognostic biomarker of human tumors.

Pan-cancer landscape of CD274 (PD-L1) copy number changes in 244 584 patient samples and the correlation with PD-L1 protein expression

IntroductionSeveral studies have shown clinical outcomes data that support the use of CD274 (PD-L1) copy-number (CN) gains and/or losses as a biomarker for immune checkpoint inhibitor (ICPI). Here, we present...

Statistical considerations for a chronic bioassay study: Exposure to Decamethylcyclopentasiloxane (D5) and incidence of uterine endometrial adenocarcinomas in a 2-year inhalation study with Fischer rats

Decamethylcyclopentasiloxane (D5) is a cyclic siloxane used in the production of industrial and consumer products. Four groups of 60 Fischer-344 female rats were analyzed for uterine endometrial adenocarcinoma (inhalation study with exposure levels in ppm/number of observed cases: 0/0, 10/1, 40/0, and 160/5) by exact regression (logistic, Poisson), the max poly-3 trend test, and a random effects probit model. When comparing the 160 ppm group to controls after 24 months, the incidence of adenocarcinomas was elevated (borderline significant); it was significant when all exposure levels were included. Four sets of (historical) control groups were formed, with varying heterogeneity. The effect of D5 was either significant or borderline significant when comparing all control sets to the 160 ppm group. When considering all exposure groups using any of the analysis methods, a significant effect was observed when the high dose group was included in the analysis; the effect was not significant when the high dose group was not included. The evidence tends to support the conclusion that D5 at the highest dose level (160 ppm) results in an increased incidence of adenocarcinomas. However, it is important to verify any potential effect through a biological investigation.

GATA3 Is a Sensitive and Specific Marker of Benign and Malignant Mesonephric Lesions in the Lower Female Genital Tract.

GATA3 is a transcription factor critical for embryogenesis, development, and cell differentiation. Recent studies have suggested that GATA3 is a sensitive and relatively specific biomarker for urothelial and breast carcinomas, with most Müllerian carcinomas being negative. We investigated GATA3 expression in mesonephric/Wolffian remnants and tumors in the female genital tract. A western blot was performed to assess specificity for the GATA3 antibody. GATA3 immunohistochemistry was performed on 59 formalin-fixed paraffin-embedded mesonephric samples, including 17 mesonephric remnants (MR; 11 cervical and 6 fallopian tube), 15 mesonephric hyperplasias, 21 mesonephric carcinomas, and 6 female adnexal tumors of probable Wolffian origin. Thirty conventional endocervical adenocarcinomas (ENDO-CA), 9 gastric-type cervical adenocarcinomas, and 165 endometrial adenocarcinomas (EM-CA) were also evaluated. GATA3 nuclear intensity and extent of staining was evaluated. The western blot revealed GATA3 expression in seminal vesicle and cell lines derived from breast and urothelial carcinomas, but not in other cell lines including ovarian, cervical, and endometrial cancers. All cervical MRs and mesonephric hyperplasias, 5/6 (83%) fallopian tube MRs, and 20/21 (95%) mesonephric carcinomas were GATA3 positive, although with great variability in both intensity (weak to strong) and extent (1+ to 3+) of staining. Only 1/6 (17%) female adnexal tumors of probable Wolffian origin showed weak multifocal staining. One of 30 (3%) usual-type ENDO-CAs and 3/165 EM-CAs exhibited weak-moderate GATA3 immunoreactivity; all gastric-type cervical adenocarcinomas were negative. GATA3 is a highly sensitive and specific marker for mesonephric lesions in the lower genital tract; however, its utility in the upper genital tract may be more limited. In addition, GATA3 can aid in distinguishing lower genital mesonephric lesions from usual-type and gastric-type ENDO-CAs and uterine EM-CAs.

Chronic toxicity and oncogenicity of decamethylcyclopentasiloxane in the Fischer 344 Rat

Decamethylcyclopentasiloxane (D5) is a cyclic polydimethylsiloxane used in the synthesis of silicon-based materials and as a component in consumer products. Male and female Fischer 344 rats were exposed to D5 vapor (0, 10, 40, 160 ppm; whole-body inhalation) for 6 h/d, 5 d/wk, for up to 104 weeks. Microscopic examination of tissues revealed test article effects at 160 ppm in the upper respiratory tract (hyaline inclusions in males and females at 6, 12, and 24 months) and an increased incidence of uterine endometrial adenocarcinoma at 24-months. The hyaline inclusions were considered a non-adverse tissue response for lack of any other respiratory tract non-neoplastic or neoplastic changes. Uterine endometrial adenocarcinoma was not anticipated. Toxicity testing (mutagenicity/genotoxicity, acute, sub-acute and sub-chronic descriptive toxicity) performed prior to the conduct of the chronic bioassay provided no indication that the uterus was a potential target organ. The target organ and tumor type specificity (adenocarcinoma is a common spontaneous tumor in the aged Fischer 344 rat) suggests the effect is associated with estrous cycle alteration. A robust assessment of potential mode(s) of action responsible for the uterine tumors and relevance to humans is addressed in a companion manuscript (Klaunig et al., 2015).

Biological relevance of decamethylcyclopentasiloxane (D5) induced rat uterine endometrial adenocarcinoma tumorigenesis: Mode of action and relevance to humans

Decamethylcyclopentasiloxane (D5) is a cyclic siloxane used in the production and formulation of consumer products with potential exposure to manufacturing workers, consumer, and the general public. Following a combined 2-year inhalation chronic bioassay performed in Fischer 344 (F344) rats, an increase in uterine endometrial adenocarcinomas was noted at the highest concentration to which animals were exposed. No other neoplasms were detected. In this study, a dose of 160 ppm produced an incidence of 8% endometrial adenocarcinomas. Based on a number of experimental studies with D5, the current manuscript examines the biological relevance and possible modes of action for the uterine endometrial adenocarcinomas observed in the rat following chronic exposure to D5. Variable rates of spontaneous uterine endometrial adenocarcinomas have been reported for untreated F344 CrlBr rats. As such, we concluded that the slight increase in uterine endometrial adenocarcinomas observed in the D5 chronic bioassay might not be the result of D5 exposure but may be related to variability of the spontaneous tumor incidence in this strain of rat. However, if the uterine endometrial adenocarcinomas are related to D5-exposure, alteration in the estrous cycle in the aging F344 rat is the most likely mode of action. D5 is not genotoxic or estrogenic. The alteration in the estrous cycle is caused by a decrease in progesterone with an increase in the estrogen:progesterone ratio most likely induced by a decrease in prolactin concentration. Available data support that exposure to D5 influences prolactin concentration. Although the effects on prolactin concentrations in a number of experiments were not always consistent, the available data support the conclusion that D5 is acting via a dopamine receptor agonist-like mechanism to alter the pituitary control of the estrous cycle. In further support of this mode of action, studies in F344 aged animals showed that the effects of D5 on estrous cyclicity produced a response consistent with a dopamine-like effect and further suggest that D5 is accelerating the aging of the reproductive endocrine system in the F344 rat utilized in this study. This mode of action for uterine endometrial adenocarcinoma tumorigenesis is not relevant for humans.

57. Uterine endometrial adenocarcinoma with an unusual immunoprofile – A case report

This case report is of a 68 year old woman who presented with post menopausal bleeding. Uterine curettings showed an adenocarcinoma with high grade features favouring a clear cell carcinoma. Subsequent hysterectomy showed a FIGO Grade 3 adenocarcinoma with predominant areas of clear cell differentiation and minor endometrioid adenocarcinoma component. Two years later, the patient presented with lung metastases. A core biopsy of lung was obtained which showed features of an adenocarcinoma with dirty necrosis, no evidence of clear cell carcinoma and immunophenotype more suggestive of a colorectal carcinoma. The original uterine tumour was then subjected to a histological review and immunohistochemical analysis. The histological features were of a high grade adenocarcinoma with areas of clear cell differentiation. In some areas, glandular differentiation with intraluminal necrosis was observed. Immunoperoxidase markers suggested a metastatic adenocarcinoma of gastrointestinal origin, whilst the immunomarkers for mullerian tract origin were negative. Gastroscopy and colonoscopy showed no evidence of a lesion. The patient meanwhile has been commenced on oral chemotherapy. This case highlights an unusual profile for an endometrial adenocarcinoma. Negative mullerian tract markers however, do not exclude a primary endometrial adenocarcinoma. It is important to know the origin of the tumour for the purpose of choosing appropriate chemotherapy for the patient.

Napsin A is a useful marker for metastatic adenocarcinomas of pulmonary origin

To address whether napsin A is useful for identifying metastatic adenocarcinomas of pulmonary origin. Fifty-four cases of adenocarcinoma that metastasized from the lungs to various sites and 1762 cases of carcinoma from various organs were immunostained for napsin A, TTF-1, CK7, CK20 and CDX2 using tissue microarray. The expression patterns of napsin A and TTF-1 in metastatic pulmonary adenocarcinomas were compared with matched primary lung tumours. Napsin A and TTF-1 were expressed in 87.0% and 81.5% of the metastatic pulmonary adenocarcinomas, respectively. Although there was no significant difference in the positivity of napsin A and TTF-1 as a single marker in metastatic pulmonary adenocarcinomas, the expression scores for napsin A were much higher than those for TTF-1 (P<0.001). Moreover, the positivity and expression scores of napsin A in primary pulmonary adenocarcinomas were maintained in metastatic adenocarcinomas better than TTF-1. Most non-pulmonary adenocarcinomas were negative for napsin A, except for renal cell carcinomas (13.4%), ovarian adenocarcinomas (7.1%) and uterine endometrial adenocarcinomas (14.5%). In particular, clear cell adenocarcinomas of ovary (68.8%) and uterus (66.7%) frequently expressed napsin A. These data suggest that napsin A may be a useful marker for identifying metastatic adenocarcinomas of pulmonary origin in combination with TTF-1.

子宮体部原発腺癌における腹腔細胞診の意義

子宮体部原発腺癌における腹腔細胞診の意義

Role of prostaglandin E2 in the experimental L-carnitine-induced endometriosis-like inflammatory model

Purpose Ectopic implantation of endometrium leading to endometriosis has been correlated with various immune and non-immune parameters. Inflammatory cytokines including IL-2, IFN-γ, TNF-α, increase in serum and peritoneal fluid of endometriotic patients, while elevated levels of prostaglandins PGE1 and PGE2 have been accused of facilitating the inflammation process. Using the experimental model of L-carnitine (L-Cn)-induced endometriosis, the aim of this study was to explore the role of PGE1 and PGE2 in endometriosis development and define whether L-Cn had a direct effect on endometrial cells. Methods During treatment with L-Cn, experimental mice were injected with indomethacin or specific PGE1 and PGE2 analogs and their effect on the L-Cn-induced inflammatory state was examined. Primary uterine cells or human endometrial adenocarcinoma cell lines were used to assess the direct or indirect effect of L-Cn. Results The seven-day treatment of virgin females with L-Cn induced obvious endometrial abnormalities with accumulation of CD90 and CD11 cells. Administration of indomethacin alleviated some of the L-Cn-induced effects, but could not inverse the phenotype. Interestingly, 11-deoxy-PGE1 aggravated the L-Cn-induced effects, whereas 17-phenyl-trimor-PGE2 reversed them. In vitro experiments showed that the effect of L-Cn depended on the nature of endometrioma cells. L-Cn could aggravate the inflammatory state of Ishikawa-inoculated mice, but could alleviate symptoms from MFE-319 inoculated females. Conculsions Although PGE2 appeared to play a key role in the L-Cn-induced uterus inflammation, the actual role of L-Cn depends on the nature of endometrioma cells and can aggravate or protect from inflammation according to the induced regulatory pathways.

In Papanicolaou smears, benign appearing endometrial cells bear no significance in predicting uterine endometrial adenocarcinomas

Reporting of benign appearing endometrial cells (BECs) in the Papanicolaou smears of women aging 40 years or older was mandated in the Bethesda System 2001 aiming at predicting the uterine pathology. The purpose of this study was to determine the clinical significance of the BECs in patients in our Medical Center. A two-arm study was designed in ≥40-years-old women with BECs and without BECs in their Pap smears from January 2002 to December 2004. Of 21,965 patients, 882 had BECs in their Pap smears and the rest did not. Among the patients with BECs, 186 (study group) and among those without BECs, 2,064 (control group) had histopathologic follow-ups. There were 4 patients in the study and 47 in the control groups who had uterine adenocarcinoma. The Chi-square P-value for adenocarcinoma between the two groups was 0.91; indicating insignificant differences between the two groups. We conclude that presence of BECs in the Pap smears of ≥40-years-old women signal no significance as a harbinger of endometrial adenocarcinoma.

Transcriptional Regulation of Aldehyde Dehydrogenase 1A1 Gene by Alternative Spliced Forms of Nuclear Factor Y in Tumorigenic Population of Endometrial Adenocarcinoma

High activity of aldehyde dehydrogenase (ALDH) is characteristic of normal and cancerous stem cells. Recently, high ALDH expression was shown to be associated with poor prognosis in uterine endometrial adenocarcinoma. The population with high ALDH activity (ALDH-hi) was more invasive, anti-apoptotic, and tumorigenic than that with low activity (ALDH-lo). Here, the transcriptional regulation of ALDH1A1 gene, which is responsible for ALDH activity, was examined in endometrial adenocarcinoma. The promoter region of ALDH1A1 contained CCAAT and octamer binding motifs, and their mutation diminished promoter activity. Among CCAAT-recognizing transcription factors, nuclear factor YA (NFYA) was involved in ALDH1A1 transcription. Two alternatively spliced isoforms of NFYA (NFYA-long and NFYA-short) have been reported. The sorted ALDH-hi population of endometrial adenocarcinoma preferentially expressed NFYA-short, whereas ALDH-lo dominantly expressed NFYA-long. NFYA-short possessed higher transactivation ability than did NFYA-long. In addition, an additive effect of NFYA with Oct-1, which recognizes octamer binding motif, was observed in ALDH1A1 transactivation. These results indicate that the alternatively spliced isoforms of NFYA, in cooperation with Oct-1, play an important role in ALDH1A1 expression in endometrial adenocarcinoma.

Inhibin/activin betaE-subunit in uterine endometrioid adenocarcinoma and endometrial cancer cell lines: From immunohistochemistry to clinical testing?

Objective Inhibins and activins are important regulators of the female reproductive system and have also been described to be involved in gynecologic cancer development. A new inhibin/activin subunit betaE has been identified, but only limited data on its expression in human uterine adenocarcinomas and endometrial cancer cell lines exist. Methods A series of 223 uterine endometrial adenocarcinomas were immunohistochemically analyzed with a specific antibody against the inhibin betaE-subunit. In addition, established endometrial cancer cell lines were analyzed for the synthesis of the inhibin betaE subunit by immunofluorescence and RT-PCR analysis. Results In this analysis, the inhibin betaE staining intensity was associated with histological grading along with a significant decrease in cases with ovarian invasion. However, inhibin-betaE did not affect patient survival nor did it constitute an independent prognostic parameter in endometrial adenocarcinoma patients. Additionally, the inhibin/activin betaE-subunit was found to be expressed in the human endometrial carcinoma cell lines HEC1a, HEC1b, Ishikawa, and RL95-2, although the level of expression was found to be highly differing among the cancer cell lines tested. Conclusion The isolated analysis of the inhibin betaE subunit might be of minor prognostic value in identifying high-risk patients. However, its differential expression in cancer of different histological differentiation and ovarian metastasis suggests a putative but yet undefined role in endometrial carcinogenesis. Whether this novel and not well studied inhibin subunit has a substantial function in the pathogenesis and malignant transformation of human endometrium is still under investigation.

Diverse facets of cervical adenocarcinoma: comprehensive review of clinicopathologic features and diagnostic criteria

Cervical adenocarcinomas span a diverse group of tumours in terms of their pathogenesis, morphology and patients’ prognosis. The diagnostic challenges are numerous and arise with virtually every case. The issues that may be particularly difficult in histopathologic evaluation of cervical adenocarcinomas include 1) differentiating in-situ and invasive tumours from their benign mimics, 2) differentiating adenocarcinoma in-situ from an early invasive lesion, 3) measuring the depth of invasion, 4) differentiating primary cervical adenocarcinoma from uterine endometrial adenocarcinoma or tumours metastatic from other primary sites such as colon. The paper reviews the pathogenesis, clinical characteristics, diagnostic criteria, immunohistochemical markers, differential diagnosis and prognosis of various subtypes of cervical adenocarcinoma.

P53 alteration independently predicts poor outcomes in patients with endometrial cancer: A clinicopathologic study of 131 cases and literature review

Objective The aim of this study was to evaluate the prognostic impact of p53 alteration in human uterine endometrial adenocarcinoma. Methods One hundred and thirty-one patients with primary endometrial adenocarcinoma were included in the study. The p53 mutation and/or protein expression were evaluated by polymerase chain reaction-single-strand conformational polymorphism and by immunohistochemical analysis, respectively. Clinical and pathological parameters were obtained from medical records. Survival data were estimated using Kaplan–Meier estimates and compared with the log-rank test where indicated. Multivariate analysis was performed using the Cox regression method. Results Thirty nine cases (29.8%) containing p53 alterations had a lower disease specific-survival rate and disease-free survival rate than those without p53 alterations. Statistically significant correlations were seen between p53 alteration and non-endometrioid histology type, high grade tumors, and the absence of progesterone receptor. Multivariate analyses showed that both p53 alteration and FIGO stage at diagnosis were adverse prognostic factors. The group of women with p53 alteration had an 11.0-fold increased risk of disease specific death (95% confidence interval: 1.008–120.765) compared to women whose tumors lacked p53 alteration. Conclusion p53 alteration defines a subset of endometrial adenocarcinoma with highly aggressive behavior and predicts lower survival in patients with endometrial adenocarcinoma.

Apoptotic effects of Tian-Long compound on endometrial adenocarcinoma cells in vitro

The Tian-Long (TL) compound is a water-soluble extract of six Chinese medicinal herbs. To explore its antitumor properties and the mechanism for activity in gynecological malignancies, the present studies were carried out using Ishikawa cells derived from uterine endometrial adenocarcinoma. Morphologically, cell death and decrease in the number of viable cells were observed in the presence of the TL compound. The proliferation of Ishikawa cells was significantly suppressed in a time- and dose-dependent manner, as indicated by both the WST-1 and the BrdU incorporation assay. Results from both the WST-1 and the BrdU incorporation assay demonstrated that the compound could inhibit the cell proliferation despite the presence of 17beta-estradiol in the medium. It is generally noted that the disturbance in mitochondrial function and DNA synthesis during cell proliferation can result in apoptosis. Being consistent with this notion, redistribution of the plasma membrane phosphatidylserine was identified with fluoromicroscopy and flow cytometry. Analysis of the fluorescent patterns of JC-1 staining revealed depolarization of the mitochondrial membrane in the exposed cells. Moreover, the amount of Bcl-2 enhanced in the presence of 17beta-estradiol was repressed by the compound. The present results indicate that the ingredients of TL compound are very promising for use in the treatment of endometrial adenocarcinoma. Further studies are necessary to elucidate the therapeutic mechanisms in its antitumor activity.

The Genomic Response of a Human Uterine Endometrial Adenocarcinoma Cell Line to 17α-Ethynyl Estradiol

We have determined the gene expression profile induced by 17 alpha-ethynyl estradiol (EE) in Ishikawa cells, a human uterine-derived estrogen-sensitive cell line, at various doses (1 pM, 100 pM, 10 nM, and 1 microM) and time points (8, 24, and 48 h). The transcript profiles were compared between treatment groups and controls (vehicle-treated) using high-density oligonucleotide arrays to determine the expression level of approximately 38,500 human genes. By trend analysis, we determined that the expression of 2560 genes was modified by exposure to EE in a dose- and time-dependent manner (p </= 0.0001). The annotation available for the genes affected indicates that EE exposure results in changes in multiple molecular pathways affecting various biological processes, particularly associated with development, morphogenesis, organogenesis, cell proliferation, cell organization, and biogenesis. All of these processes are also affected by estrogen exposure in the uterus of the rat. Comparison of the response to EE in both the rat uterus and the Ishikawa cells showed that 71 genes are regulated in a similar manner in vivo as well as in vitro. Further, some of the genes that show a robust response to estrogen exposure in Ishikawa cells are well known to be estrogen responsive, in various in vivo studies, such as PGR, MMP7, IGFBP3, IGFBP5, SOX4, MYC, EGR1, FOS, CKB, and CCND2, among others. These results indicate that transcript profiling can serve as a viable tool to select reliable in vitro systems to evaluate potential estrogenic activities of target chemicals and to identify genes that are relevant for the estrogen response.

PIK3CA mutations in the kinase domain (exon 20) of uterine endometrial adenocarcinomas are associated with adverse prognostic parameters

Mutations of the oncogene PIK3CA occur frequently in endometrial carcinomas, but their prognostic significance is unclear. To determine the clinicopathological and molecular implications of these mutations, PIK3CA status was investigated in 109 endometrial (102 endometrioid and 7 mixed) carcinomas and the results were compared with clinicopathological parameters associated with prognosis. Tumors were also investigated for microsatellite instability and PTEN, β-catenin gene (CTNNB1), K-RAS, and B-RAF mutations. We found 35 PIK3CA somatic missense mutations in 32 (29%) endometrial carcinomas. Eighteen mutations occurred in exon 20 (kinase domain), and 17 in exon 9 (helical domain). Almost all mutated tumors were pure endometrioid adenocarcinomas. All tumors with PIK3CA mutations exhibited myometrial invasion (P=0.032). Lymphovascular invasion was found more frequently in mutated (28%) than nonmutated carcinomas (18%). Histological grade varied significantly according to the location of the PIK3CA mutations whether in exon 9 or exon 20 (P=0.033). The frequency of exon 9 mutations was higher in grade 1 carcinomas (57%) than in grade 2 (29%) or grade 3 (14%) tumors. Conversely, mutations in exon 20 were more common in grade 3 (60%) than in grade 2 (20%) or grade 1 (20%) carcinomas. None of the tumors confined to the endometrium (stage IA) had PIK3CA mutations. Furthermore, whereas 64% of adenocarcinomas with exon 9 mutations had invaded ≤½ of the myometrial thickness (stage IB), 73% of tumors with exon 20 mutations had either deeper myometrial invasion (stage IC) or cervical involvement (stage II) (P=0.045). PIK3CA mutations coexisted with microsatellite instability and mutations in PTEN, CTNNB1, K-RAS, and B-RAF genes. These results favor that PIK3CA mutations are associated with myometrial invasion and, moreover, that tumors harboring PIK3CA mutations in exon 20 are frequently high-grade, deeply invasive endometrial carcinomas that tend to exhibit lymphovascular invasion.