Uterine Endometrial Cancer Research Articles

The combination of Paclitaxel and Gefitinib inhibits endometrial cancer cells by inducing mitotic catastrophe: proof of principle for dual therapy in endometrial cancer

Serous uterine endometrial cancer is a lethal disease for which new therapeutic regimens are urgently needed. Combinations of chemotherapeutic agents and small molecule growth factor inhibitors have demonstrated activity in cancers from other sites. Our objective was to determine whether such a combination using Paclitaxel and Gefitinib could be active in serous endometrial cancer cells.

The cases of Concurrent Chemo-Photodynamic therapy (CCPDT) for the uterine endometrial cancer in young women desiring fertility

The cases of Concurrent Chemo-Photodynamic therapy (CCPDT) for the uterine endometrial cancer in young women desiring fertility

Expression ratio of CCND1 to CDKN2A mRNA predicts RB1 status of cultured cancer cell lines and clinical tumor samples

BackgroundThe retinoblastoma product (RB1) is frequently deregulated in various types of tumors by mutation, deletion, or inactivation through association with viral oncoproteins. The functional loss of RB1 is recognized to be one of the hallmarks that differentiate cancer cells from normal cells. Many researchers are attempting to develop anti-tumor agents that are preferentially effective against RB1-negative tumors. However, to identify patients with RB1-negative cancers, it is imperative to develop predictive biomarkers to classify RB1-positive and -negative tumors.ResultsExpression profiling of 30 cancer cell lines composed of 16 RB1-positive and 14 RB1-negative cancers was performed to find genes that are differentially expressed between the two groups, resulting in the identification of an RB1 signature with 194 genes. Among them, critical RB1 pathway components CDKN2A and CCND1 were included. We found that microarray data of the expression ratio of CCND1 and CDKN2A clearly distinguished the RB1 status of 30 cells lines. Measurement of the CCND1/CDKN2A mRNA expression ratio in additional cell lines by RT-PCR accurately predicted RB1 status (12/12 cells lines). The expression of CCND1/CDKN2A also correlated with RB1 status in xenograft tumors in vivo. Lastly, a CCND1/CDKN2A assay with clinical samples showed that uterine cervical and small cell lung cancers known to have a high prevalence of RB1-decifiency were predicted to be 100% RB1-negative, while uterine endometrial or gastric cancers were predicted to be 5-22% negative. All clinically normal tissues were 100% RB1-positive.ConclusionsWe report here that the CCND1/CDKN2A mRNA expression ratio predicts the RB1 status of cell lines in vitro and xenograft tumors and clinical tumor samples in vivo. Given the high predictive accuracy and quantitative nature of the CCND1/CDKN2A expression assay, the assay could be utilized to stratify patients for anti-tumor agents with preferential effects on either RB1-positive or -negative tumors.

Effects of a selective COX-2 inhibitor in patients with uterine endometrial cancers

COX-2 is highly expressed in endometrial cancers, suggesting that a selective COX-2 inhibitor could be valuable for treating endometrial cancers that overexpress COX-2. In this study, we investigated the anti-tumor effects of the selective COX-2 inhibitor etodolac on endometrial cancer patients. Etodolac (400 mg, bid, for 2 weeks) was administered preoperatively to 21 endometrial cancer patients who had provided informed consent. Using pre-treatment biopsies and post-treatment surgical specimens, the expression levels of COX-2, Ki-67, p53, p21, p27, and cyclin D1 were evaluated by immunohistochemistry and the apoptotic index (AI) was determined by TUNEL staining. Preoperative biopsies and surgical specimens from 32 patients with endometrial cancer not treated with etodolac served as controls. Surgical specimens from COX-2 positive endometrial cancer patients treated with etodolac had significantly reduced expression levels of COX-2, Ki-67, p53, p21, p27, and cyclin D1 as determined by immunohistochemistry, while AI was not affected. These markers were unchanged for COX-2 negative endometrial cancer patients treated with etodolac and the control group. The selective COX-2 inhibitor etodolac showed anti-proliferative effects by suppressing COX-2 and cell-cycle regulator protein expression in patients with endometrial cancer positive for COX-2 expression. This study demonstrates that a selective COX-2 inhibitor is a potentially beneficial treatment for COX-2 positive endometrial cancers.

Sex steroids in uterine endometrial cancers.

Some uterine endometrial cancers conserve estrogen dependency in advancement. However, the concept of advancement in tumor is complicated, because it involves simple growth in primary tumor and secondary spreading. The expression manner of estrogen receptor alpha exon 5 splicing variant, ER beta, progesterone receptor-A (N-terminus deletion mutant) is associated with metastatic potential in uterine endometrial cancers. Increased estrogen-related receptor alpha expression is related to tumor advancement with the loss of estrogen dependency. Steroid receptor coactivator-3 contributes to tumor progression and can be used as a treatment target for advanced uterine endometrial cancers. Estrogen responsive oncogenes, c-jun and c-Ha-ras, are not modi-fied by progestin in uterine endometrial cancer cells and are considered to be an instinct phenotype as such cancers. By contrast, metastatic potential of estrogen-dependent uterine endometrial cancers can be partially controlled by progestin via metastasis-related genes, E-cadherin/catenins, plasminogen activator inhibitor-1, vascular endothelial growth factor. Thus, sex steroids related phenomena are impress-ive in the advancement of uterine endometrial cancers.

Use of Localization and Activity of Thymidine Phosphorylase in Human Gynecological Tumors for Predicting Sensitivity to Pyrimidine Antimetabolite Therapy: An Observational Study

Background: Thymidine phosphorylase (dThdPase) is the rate-limiting enzyme in the conversion of 5 ′ -deoxy- 5-fluorouridine (5 ′ -dFUrd), an intermediate metabolite of capecitabine (Xeloda ® ), to 5-fluorouracil (5-FU). We investigated the correlation between dThdPase activity and immunohistochemical staining in gynecological carcinoma and adjacent normal tissues. We hypothesize that the differential dThdPase activity between tumors and adjacent tissue is predictive of response to treatment with pyrimidine antimetabolites. Methods: In 45 samples of carcinoma tissue and 35 of adjacent normal tissue from 45 patients, we measured dThdPase activity as well as immunoreactivity using an anti-dThdPase monoclonal antibody and macrophage and histiocyte-specific antibodies. Results: dThdPase activity in tumor tissue was significantly higher than that in the corresponding adjacent normal tissue in all samples tested (12 uterine cervical, 19 endometrial, and 4 ovarian tumors). Anti-dThdPase immunopositivity was observed in the epithelial tumor cells of 76.9% of uterine cervical cancer samples, 60.0% of endometrial cancer samples and 63.6% of ovarian cancer samples. In stromal tissue, 84.6% of uterine cervical tumors (11/13), 90.0% of endometrial tumors (18/20), and 81.8% of ovarian tumors (9/11) were immunopositive for anti-dThdPase in interstitial cells (mainly macrophages). Macrophages were also strongly reactive in the stromal tissues of uterine cervical, endometrial, and ovarian cancers. The correlation between dThdPase activity and intensity of immunohistochemical staining of epithelial tumor cells with anti-dThdPase monoclonal antibody was statistically significant in endometrial carcinoma ( P = 0.008) but borderline in uterine cervical tumors ( P = 0.077). We found a good correlation between dThdPase activity and staining of epithelial tumor cells, particularly in the case of endometrial cancer. Conclusions: We show that gynecological carcinomas show increased dThdPase activity, and this activity correlates with dThdPase staining of tumor epithelial cells. Thus, dThdPase staining of biopsy specimens could be useful in predicting the outcome of therapy with pyrimidine metabolites.

Evaluation of Parametrial Spread in Endometrial Carcinoma

To evaluate the detailed clinicopathologic characteristics of parametrial spread in uterine endometrial cancer. We retrospectively identified 334 individuals with uterine endometrial cancer who had undergone radical hysterectomy between 1988 and 2007. Parametrial spread was determined by histopathological analysis of surgically resected specimens. Twenty-eight (8.4%) individuals had histopathologically confirmed parametrial spread, and lymphatic or blood vessel invasion (22 cases) was the most frequently observed type of parametrial spread; direct invasion to parametrial connective tissue (five cases) or cardinal lymph node metastasis (four cases) were less frequently observed. Parametrial spread occurred not only in individuals with cervical involvement but also in individuals with more than half myometrial invasion, retroperitoneal (pelvic, paraaortic, or both), lymph node metastasis, ovarian metastasis, positive peritoneal cytology results, and lymphovascular space invasion. Twenty-six individuals (92.9%) with parametrial spread showed more than one of these histopathological factors (median number of factors 3, range 1-6); the other two individuals had lymphovascular space invasion alone. In 10 individuals with parametrial spread (35.7%), the condition recurred during the median follow-up period of 49 months, and initial recurrence was observed in the lung in six individuals (60.0%). Although the long-term prognosis for those with parametrial spread was significantly poorer than that of those without parametrial spread, both among all individuals (P<.001) and among individuals with International Federation of Gynecology and Obstetrics stage III (P<.05), multivariate analysis showed that parametrial spread was not an independent prognostic factor for uterine endometrial cancer. Parametrial spread cannot be predicted by cervical involvement alone but may be predicted by various lymphovascular space invasion-related histopathologic factors. Further, parametrial spread may not be an independent prognostic factor in individuals with uterine endometrial cancer. III.

Lipid Peroxidation in Erythrocyte Membranes of Women with Benign and Malignant Neoplasms of the Endometrium

The content of primary and secondary products of membrane peroxidation, total cholesterol concentration, and relative microviscosity of erythrocyte membranes were measured in women with endometrial cancer and uterine leiomyoma. The content of conjugated dienes in erythrocytes was highest in female patients with malignant tumors of the endometrium. The relative microviscosity of erythrocyte membranes in patients with uterine leiomyoma and endometrial cancer was elevated in the zone of lipid--lipid and protein--lipid contacts.

Abstract 1795: The Combination of Paclitaxel and Gefitinib Inhibits Endometrial Cancer Cells by Inducing Mitotic Catastrophe: Proof of Principle for Dual Therapy in Endometrial Cancer

Abstract Serous uterine endometrial cancer is a lethal disease for which new therapeutic regimens are urgently needed. Combinations of chemotherapeutic agents and small molecule growth factor inhibitors have demonstrated activity in cancers from other sites. Our objective was to determine whether such a combination could be active in serous endometrial cancer cells. The effects of the EGFR inhibitor gefitinib (ZD1839, Iressa) alone, paclitaxel alone, and the combination of both agents on the cell cycle and on cell proliferation were studied using Hec50co cells, a validated model for type II serous endometrial cancer with absent p53. First, we established the IC50 for paclitaxel alone (14 nM) compared to that of paclitaxel and gefitinib in combination (1.3 nM). This 10-fold reduction in the IC50 with dual therapy yielded a combination index of 0.25, strongly suggesting that the paclitaxel and gefitinib combination resulted in synergistic growth inhibition. Sixty-seven percent of the cells treated with paclitaxel and gefitinib arrested in the G2/M phase. This arrest of the cell cycle at G2/M with combination therapy was significantly greater than with single agent treatment alone (p &amp;lt; 0.001) and was associated with inhibition of p38 phosphorylation and the induction of CDC25C phosphatase and Cyclin B1. When induced, these factors abrogate the G2/M checkpoint and result in rapid, premature progression to M phase; this mechanism is particularly active in the setting of absent p53. With rapid progression into M, the cells were highly sensitive to paclitaxel and were killed by the mechanism of mitotic catastrophe. Similar findings were observed in the same cells treated with the combination of paclitaxel and a specific inhibitor of p38, SB203580. Our study suggests that inhibition of EGFR or downstream p38 pathway can abrogate the mitotic stress checkpoint induced by cytotoxic agents targeting microtubules such as paclitaxel. These findings suggest a new therapeutic strategy for the treatment of serous endometrial carcinoma worthy of clinical confirmation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1795.

MicroRNA regulation of the expression of the estrogen receptor in endometrial cancer

microRNAs (miRNAs) are non-coding RNA transcripts thought to be instrumental in controlling eukaryotic cell function. This study aimed to identify miRNAs associated with the differential expression of estrogen receptor α (ERα) and progesterone receptor (PR) in uterine endometrial cancer. First, the pathology of Ishikawa and KLE cell lines was identified by transplant and biopsy. Expression levels of ERα and PR and their response to estrogen and progesterone were examined, and total RNA was isolated to identify differentially expressed miRNAs by microarray analysis. miRNAs targeting ESR1 and PGR were predicted by miRANDA and TargetScan software, and their expression levels in cell lines and in patient samples were validated by real-time RT-PCR. Ishikawa and KLE represent estrogen-dependent and estrogen-independent endometrial cancers, respectively. As identified by miRNA microarray, 126 miRNAs were differentially expressed between the two cell lines; hsa-miR-100 and hsa-miR-99a were predicted to target ESR1, and hsa-miR-378 and hsa-miR-768-3p to target PGR. The differential expression of these miRNAs between Ishikawa and KLE was consistent in vivo and in vitro. Hsa-miR-100 was significantly down-regulated in the estrogen-dependent endometrial cancer samples as compared to the estrogen-independent samples and thus has the potential to target ESR1.

Role of inhibitor of DNA binding-1 protein is related to angiogenesis in the tumor advancement of uterine endometrial cancers

The inhibitor of DNA binding (ID)-1 protein, an inhibitor of basic helix-loop-helix transcription factors, has been found to be involved in multiple cellular functions. In the present study, ID-1 histoscores and mRNA levels were both significantly (p<0.05) increased in uterine endometrial cancers according to clinical stage, histological grade and depth of myometrial invasion. Furthermore, the 60-month survival rate of the 25 patients with high ID-1 was poor (52%), while that of the other 25 patients with low ID-1 was significantly higher (80%) (p<0.05). ID-1 histoscores and mRNA levels significantly (p<0.0001) correlated with microvessel counts in uterine endometrial cancers. Therefore, ID-1 acts on tumor advancement via angiogenic activity and can be considered a candidate prognostic indicator in uterine endometrial cancers.

Tamoxifen and arrhythmia

Tamoxifen is an orally active selective estrogen receptor modulator that is used in the treatment of breast cancer and is currently the world's largest selling drug for that purpose. However, it has some side effects including hot flashes, menstrual irregularity, vaginal discharges, uterine bleeding, uterine endometrial cancer, hypercoagulability, steatosis hepatis, risk of trombembolism. Long-term data from clinical trials have failed to demonstrate a cardioprotective effect and beneficial effects on serum lipid profiles. Arrhythmia secondary to tamoxifen is very rare.

Diffusion-weighted magnetic resonance imaging of endometrial cancer: differentiation from benign endometrial lesions and preoperative assessment of myometrial invasion

Uterine endometrial cancer is the most common gynecologic malignancy, and benign endometrial hyperplasia or polyps should be differentiated from endometrial cancer. In evaluating endometrial cancer on magnetic resonance imaging (MRI), the assessment of the depth of myometrial invasion is important because it closely correlates with the patient's prognosis. To verify the feasibility of diffusion-weighted magnetic resonance imaging (DWI) to distinguish benign and malignant endometrial lesions, and to evaluate myometrial invasion of endometrial cancer. Sixty-seven endometrial lesions including 45 cancers and 22 benign lesions (hyperplasia and polyps) were evaluated by DWI with apparent diffusion coefficient (ADC) measurement. The staging accuracies of DWI and gadolinium-enhanced T1-weighted images in the assessment of myometrial invasion were evaluated in 33 patients with endometrial cancer. The ADC values (x10(-3) mm(2)/s) in cancer and benign lesions were 0.84+/-0.19 and 1.58+/-0.36, respectively (P<0.01). The staging accuracy (superficial or deep myometrial invasion) was 94% for DWI and 88% for gadolinium-enhanced T1-weighted images. Coexisting adenomyosis and infiltrative myometrial invasion caused staging errors on gadolinium-enhanced T1-weighted images, whereas DWI could demonstrate the tumor extent correctly. DWI provides helpful information in evaluating benign and malignant endometrial lesions.

FGFR2 Abnormalities Underlie a Spectrum of Bone, Skin, and Cancer Pathologies

Fibroblast growth factor receptor (FGFR)2 is regulated on the basis of the balance of FGFs, heparan-sulfate proteoglycans, FGFR2 isoforms, endogenous inhibitors, and microRNAs. FGFR2 signals cross-talk with hedgehog, bone morphogenetic protein, and other regulatory networks. Some cases of congenital skeletal disorders with an FGFR2 mutation show skin phenotypes, including acne, cutis gyrata, and acanthosis nigricans. Gain-of-function mutations or variations of human FGFR2 occur in estrogen receptor-positive breast cancer, diffuse-type gastric cancer, and endometrial uterine cancer. Oral administration of AZD2171 or Ki23057 inhibits in vivo proliferation of cancer cells with aberrant FGFR2 activation in rodent therapeutic models. However, loss-of-function mutations of FGFR2 are reported in human melanoma. Conditional Fgfr2b knockout in the rodent epidermis leads to increased macrophage infiltration to the dermis and adipose tissue, epidermal thickening accompanied by basal-layer dysplasia and parakeratosis, and the promotion of chemically induced squamous-cell carcinoma. Dysregulation of FGFR2 results in a spectrum of bone and skin pathologies and several types of cancer.

Uterine Junctional Zone and Endometrial Cancer

PURPOSE: The human uterus consists of the ontogenetically and phylogenetically older endometrium, the adjacent junctional zone and the more recent outer myometrium. This understanding of uterine zonal anatomy could provide an answer to the question why the risk of lymph node metastasis increases abruptly when an endometrial adenocarcinoma invades more than 50 % of the myometrial wall. MATERIAL AND METHODS: High resolution vaginal ultrasound was performed in 100 healthy postmenopausal women and their zonal anatomy was documented. In a meta-analysis the relationship between the depth of myometrial invasion by endometrial cancer and the rate of regional lymph node metastases was analyzed. RESULTS: In postmenopausal women the mean diameter of the junctional zone of the myometrium was found to be almost exactly half the inner diameter of the myometrium. Our meta-analysis confirmed the existence of a threshold whereby, if an endometrial adenocarcinoma invaded more than 50 % of the myometrial wall, this resulted in a more than 5-fold higher prevalence of lymph node metastasis. CONCLUSION: Endometrial cancer rarely results in lymph node metastasis as long as the disease is confined to the junctional zone of the myometrium. The dissimilarity between the inner and outer myometrial layers, based on their very different phylogenetic and ontogenetic origins, might be of interest for further research.

Evaluation of The Efficacy of Systematic Pelvic Lymphadenectomy in Endometrial Cancer

Evaluation of: The ASTEC Study Group: Efficacy of systematic pelvic lymphadenectomy in endometrial cancer (MRC ASTEC trial): a randomized study. Lancet 373, 125–136 (2009). Pelvic lymph node metastasis is one of the most common causes of extrauterine tumor spread in early-stage endometrial uterine cancer. Many retrospective studies have shown the significance of pelvic lymphadenectomy in patients with what was suspected preoperatively to be early-stage endometrial carcinoma. The study evaluated here is the largest prospective randomized trial evaluating pelvic lymphadenectomy in early-stage endometrial cancer, and provides the direct and fully reported overall and recurrence-free survival rates comparing standard surgery with standard surgery plus pelvic lymphadenectomy in patients with early-stage endometrial carcinoma. Overall, the results showed no evidence of benefit in terms of overall or recurrence-free survival for pelvic lymphadenectomy in women with early-stage endometrial cancer. However, this trial analyzed the results of overall and recurrence-free survival without considering the combination of histopathological prognostic risk factors. It may be important to analyze overall and recurrence-free survival between the subgroups by combination of the histopathological prognostic risk factors. A large, prospective clinical trial might be necessary to evaluate the role of pelvic lymphadenectomy by considering the combination of histopathological prognostic risk factors in early-stage endometrial cancer.

Clinical implication of estrogen-related receptor (ERR) expression in uterine endometrial cancers

Estrogen receptor (ER)α and ERβ mRNAs levels decreased with clinical stage, myometrial invasion and dedifferentiation. On the other hand, ERRα mRNA levels and histoscores increased with clinical stage and myometrial invasion, regardless of dedifferentiation. ERRα can bind to the steroid receptor coactivator family without any ligands, and drive transcription activity of the target genes. The competitive interaction of ERRα/ER expression associated with the use of common cofactors during loosing estrogen dependency might cause their expression manner. The up-regulation of ERRα might be related to tumor growth and advancement in uterine endometrial cancers. It is speculated that ERRα is a candidate for prognostic factors in uterine endometrial cancer, although ERRs are not directly related to growth of uterine endometrial cancer.

Homeobox gene HOPX is epigenetically silenced in human uterine endometrial cancer and suppresses estrogen‐stimulated proliferation of cancer cells by inhibiting serum response factor

HOPX (homeodomain only protein X) is a newly identified homeobox gene whose loss of expression has been reported for several types of neoplasm. Although we found most human uterine endometrial cancers (HEC) defective in HOPX expression, genetic mutations in the HOPX gene were undetectable. As is the case with several tumor suppressor genes, the promoter region of HOPX is densely methylated in HEC tissue samples obtained by laser capture microdissection. HOPX mRNA and protein levels were reduced in the majority of samples, and this correlated with hypermethylation of the HOPX promoter. Forced expression of HOPX resulted in a partial block in cell proliferation, in vivo tumorigenicity and c-fos gene expression in HEC and MCF7 cells in response to 17beta-estradiol (E(2)) stimulation. Analysis of the serum response element (SRE) of c-fos gene promoter showed that the effect of HOPX expression is associated with inhibition of E(2)-induced c-fos activation through the serum response factor (SRF) motif. Knockdown of HOPX in immortalized human endometrial cells resulted in accelerated proliferation. Our study indicates that transcriptional silencing of HOPX results from hypermethylation of the HOPpromoter, which leads to HEC development.

Large‐scale quantitative clinical proteomics by label‐free liquid chromatography and mass spectrometry

We previously reported the development of an integrated proteome platform, namely 2-Dimensional Image Converted Analysis of Liquid chromatography and mass spectrometry (2DICAL), for quantitative comparison of large peptide datasets generated by nano-flow liquid chromatography (LC) and mass spectrometry (MS). The key technology of 2DICAL was the precise adjustment of the retention time of LC by dynamic programming. In order to apply 2DICAL to clinical studies that require comparison of a large number of patient samples we further refined the calculation algorithm and increased the accuracy and speed of the peptide peak alignment using a greedy algorithm, which had been used for fast DNA sequence alignment. The peptide peaks of each sample with the same m/z were extracted every 1 m/z and displayed with along the horizontal axis. Here we report a precise comparison of more than 150,000 typtic peptide ion peaks derived from 70 serum samples (40 patients with uterine endometrial cancer and 30 controls). The levels of 49 MS peaks were found to differ significantly between cancer patients and controls (P < 0.01, Welch's t-test and interquartile range [IQR] of >40), and the differential expression and identification of selected three proteins was validated by immunoblotting. 2DICAL was is highly advantageous for large-scale clinical proteomics because of its simple procedure, high throughput, and quantification accuracy.

Novel therapeutic strategy for uterine endometrial cancers

In general, tumors induce angiogenic factors specific to them, which leads to angiogenesis with tumor progression. However, angiogenesis in uterine endometrial cancers is complicated because the hormone dependency in their growth also modifies the angiogenic potential. Therefore, angiogenic potential in uterine endometrial cancers must be thoroughly analyzed. The upstream of the vascular endothelial growth factor (VEGF) gene conserves estrogen-responsive elements. Progesterone primed with estrogen induces thymidine phosphorylase (TP) in the uterine endometrium. Sex steroid-dependent VEGF and TP are highly expressed in early-stage and well-differentiated uterine endometrial cancers, and basic fibroblast growth factor (bFGF) is highly expressed in advanced and poorly differentiated uterine endometrial cancers. A transcriptional factor for angiogenesis, E26 transformation specific (ETS-1), is linked to VEGF in well-differentiated uterine endometrial cancers, and to bFGF in poorly differentiated uterine endometrial cancers. Therefore, even if dedifferentiation and angiogenic switching occur due to tumor progression and long-term hormone therapy, the inhibition of ETS-1, along with inhibition of the main angiogenic factors, may be an effective strategy to suppress uterine endometrial cancers as a novel antiangiogenic therapy.