Cíl studie: Stanovit endometriální profi ly exprese mesengerové RNA (mRNA) vybraných matrixmetaloproteináz (MMP) a tkáňového inhibitoru metaloproteináz-1 (TIMP-1).Typ studie: Experimentální studie.Název a sídlo pracoviště: Gynekologicko-porodnická klinika, LF UP a FN Olomouc, Česká republika,Gynekologicko-porodnická klinika, Univerzitní nemocnice Lund, Švédsko.Metodika: Studovali jsme MMP-1, -3, -7, -10, -11, -12, -13, -14, -16, -26 a TIMP-1 mRNA ve 39 vzorcíchnormální endometriální tkáně získaných z různých fází menstruálního cyklu. Na základě histologickéhovyšetření byly vzorky klasifi kovány jako časná (n=8), střední (n=6) a pozdní (n=7) proliferace,časná (n=4), střední (n=4) a pozdní (n=8) sekrece a menstruální (n=3) fáze. Hladiny mRNAextrahované ze zmražených tkáňových vzorků byly kvantifi kovány pomocí real time PCR.Výsledky: Celkem byly zjištěny čtyři vzorce exprese MMP mRNA v průběhu menstruačního cyklu.MMP-1, -3, -10, a -12 byly exprimovány převážně v perimenstruálním období, MMP-7 a -11 mělynejvyšší hladiny v proliferační fázi, MMP-13, -14 a -16 byly exprimovány v průběhu celého cyklua MMP-26 měla maximum exprese v periovulatorním období. Hladiny TIMP-1 mRNA zůstávalynezměněny během cyklu.Závěr: Specifi cké profi ly endometriální exprese MMP v průběhu menstruačního cyklu ukazují najejich odlišný biologický význam v průběhu cyklu. MMP-26 vykazuje samostatný vzorec exprese.
Objective: To examine the endometrial expression pattern of messenger RNA (mRNA) for selectedmatrix metalloproteinases (MMP) and tissue inhibitor of metalloproteinases-1 (TIMP-1).Design: Experimental study.Setting: Department of Obstetrics and gynecology of the Palacky University Medical School andUniversity Hospital, Olomouc, Czech Republic, Department of Obsterics and Gynecology, UniversityHospital, Luna, Sweden.Methods: We studied MMP-1, -3, -7, -10, -11, -12, -13, -14, -16, -26 and TIMP-1 mRNA in 39 normalendometrial samples obtained across the menstrual cycle. Based on histological examination, allspecimens were classifi ed according to an ideal 28 day menstrual cycle as early (n=8), mid (n=6)and late (n=7) proliferative phase, early (n=4), mid (n=4) and late (n=8) secretory phase and menstrual(n=3) phase. mRNA extracted from frozen tissue samples was quantitated using real timePCR.Results: Four distinct patterns of MMP mRNA expression were detected in cycling endometrium.MMP-1, -3, -10, and -12 were expressed predominantly in perimenstrual period, MMP-7 and -11had highest levels in proliferative phase, MMP-13, -14 and -16 were expressed throughout thecycle and MMP-26 was found to be maximal in periovulatory period. Levels of TIMP-1 mRNAremained unchanged during the cycle.Conclusion: The specifi c endometrial expression profi les of MMPs during menstrual cycle pointto their specifi c biologic roles during the cycle. MMP-26 exhibits a unique expression pattern.
BACKGROUND/AIM: The presence of circulating tumor cells (CTCs) in the peripheral blood of patients with solid tumors is associated with a poor prognosis. However, there are limited data concerning the detection of CTCs in endometrial cancer (EC). The aim of this study was to evaluate the presence of CTCs in the peripheral blood of patients with EC. MATERIALS AND METHODS: Peripheral blood samples from 92 patients who underwent a surgical procedure were evaluated using MetaCell® separation technology for CTCs. RESULTS: CTCs were detected in 69 (75%) patients with EC. CONCLUSION: CTCs were detected in a higher percentage of patients than in other studies. The results showed that the technology applied in this study can efficiently capture viable tumor cells in the blood that can be cultured while maintaining their original phenotype. This paper discusses the first successful culturing of human circulating endometrial cancer cells for further downstream functional and molecular characterization.
- MeSH
- Humans MeSH
- Neoplastic Cells, Circulating pathology MeSH
- Endometrial Neoplasms blood pathology MeSH
- Neoplasm Staging MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
OBJECTIVE: To evaluate whether the amount of preoperative endometrial tissue surface is related to the degree of concordance with final low- and high-grade endometrial cancer (EC). In addition, to determine whether discordance is influenced by sampling method and impacts outcome. METHODS: A retrospective cohort study within the European Network for Individualized Treatment of Endometrial Cancer (ENITEC). Surface of preoperative endometrial tissue samples was digitally calculated using ImageJ. Tumor samples were classified into low-grade (grade 1-2 endometrioid EC (EEC)) and high-grade (grade 3 EEC + non-endometroid EC). RESULTS: The study cohort included 573 tumor samples. Overall concordance between pre- and postoperative diagnosis was 60.0%, and 88.8% when classified into low- and high-grade EC. Upgrading (preoperative low-grade, postoperative high-grade EC) was found in 7.8% and downgrading (preoperative high-grade, postoperative low-grade EC) in 26.7%. The median endometrial tissue surface was significantly lower in concordant diagnoses when compared to discordant diagnoses, respectively 18.7 mm2 and 23.5 mm2 (P = 0.022). Sampling method did not influence the concordance in tumor classification. Patients with preoperative high-grade and postoperative low-grade showed significant lower DSS compared to patients with concordant low-grade EC (P = 0.039). CONCLUSION: The amount of preoperative endometrial tissue surface was inversely related to the degree of concordance with final tumor low- and high-grade. Obtaining higher amount of preoperative endometrial tissue surface does not increase the concordance between pre- and postoperative low- and high-grade diagnosis in EC. Awareness of clinically relevant down- and upgrading is crucial to reduce subsequent over- or undertreatment with impact on outcome.
- MeSH
- Biopsy methods MeSH
- Endometrium pathology MeSH
- Carcinoma, Endometrioid * surgery pathology MeSH
- Humans MeSH
- Endometrial Neoplasms * pathology MeSH
- Retrospective Studies MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Hepatocyte nuclear factor 1-beta (HNF-1-beta) is a transcription factor involved in cancerogenesis of various tumors, including endometrioid carcinoma. We performed comprehensive analysis of HNF-1-beta in lesions of the endometrium, including protein expression and genetic and epigenetic changes. Expression of HNF-1-beta was analyzed immunohistochemically in 320 cases including both tumor and non-tumor endometrial lesions. Promoter methylation and genetic variants were evaluated, using bisulphite and direct sequencing, in 30 (18 fresh frozen, 12 FFPE tumors) endometrioid carcinomas (ECs) and 15 ovarian clear cell carcinomas (OCCCs) as a control group. We detected expression of HNF-1-beta in 28 % of ECs (51/180 cases), 26 % of serous carcinoma (7/27 cases), 83 % of endometrial clear cell carcinoma (15/18 cases), 93 % of hyperplastic polyps with atypias (13/14 cases), 100 % of hyperplastic polyps without atypias (16/16 cases), 88 % of hyperplasias with atypias (14/16 cases), 91 % of hyperplasias without atypias (10/11 cases), and in ≥80 % of different normal endometrium samples. The control group of OCCCs showed HNF-1-beta expression in 95 % (18/19 cases). Methylation in promoter region was detected in 13.3 % (4/30) of ECs, but not in corresponding normal tissue where available, nor in OCCCs (0/15 cases). Mutation analysis revealed truncating variant c.454C > T (p.Gln152X) in one EC and missense variant c.848C > T (p.Ala283Val) was detected in one OCCC. In conclusion, expression of HNF-1-beta was detected in various extents in all types of lesions analyzed, nevertheless its strong expression was mostly limited to clear cell carcinomas. Biological significance of genetic and epigenetic changes needs further investigation.
- MeSH
- Adenocarcinoma, Clear Cell genetics pathology MeSH
- Carcinoma, Endometrioid genetics pathology MeSH
- Epigenesis, Genetic genetics MeSH
- Epigenomics methods MeSH
- Genetic Variation genetics MeSH
- Hepatocyte Nuclear Factor 1-beta genetics MeSH
- Immunohistochemistry methods MeSH
- Humans MeSH
- DNA Methylation genetics MeSH
- Biomarkers, Tumor genetics MeSH
- Endometrial Neoplasms genetics pathology MeSH
- Promoter Regions, Genetic genetics MeSH
- Cystadenocarcinoma, Serous genetics pathology MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Phosphatase and tensin homolog (PTEN) is a protein that acts as a tumor suppressor by dephosphorylating the lipid second messenger phosphatidylinositol 3,4,5-trisphosphate. Loss of PTEN function has been implicated in the pathogenesis of a number of different tumors, particularly endometrial carcinoma (ECa). ECa is the most common neoplasia of the female genital tract. Our study evaluates an association between the morphological appearance of endometrial hyperplasia and endometrial carcinoma and the degree of PTEN alterations. A total of 45 endometrial biopsies from Slovak women were included in present study. Formalin-fixed and paraffin-embedded tissue samples with simple hyperplasia (3), complex hyperplasia (5), atypical complex hyperplasia (7), endometrioid carcinomas G1 (20) and G3 (5), and serous carcinoma (5) were evaluated for the presence of mutations in coding regions of PTEN gene, the most frequently mutated tumor suppressor gene in endometrial carcinoma. 75% of the detected mutations were clustered in exons 5 and 8. Out of the 39 mutations detected in 24 cases, 20 were frameshifts and 19 were nonsense, missense, or silent mutations. Some specimens harboured more than one mutation. The results of current study on Slovak women were compared to a previous study performed on Polish population. The two sets of results were similar.
- MeSH
- PTEN Phosphohydrolase genetics MeSH
- Endometrial Hyperplasia genetics MeSH
- Humans MeSH
- Molecular Sequence Data MeSH
- Mutation genetics MeSH
- DNA Mutational Analysis MeSH
- Mutation Rate MeSH
- Endometrial Neoplasms genetics MeSH
- Base Sequence MeSH
- Sequence Analysis, DNA * MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Slovakia MeSH
BACKGROUND: Glucose transporter-1 (Glut-1) is a membrane glycoprotein that is, together with other glucose transporters, responsible for the regulation of glucose uptake. An increased expression of this protein seems to be a general feature of several malignant tumors that are able to reprogram their metabolism and switch from oxidative phosphorylation to aerobic glycolysis. METHODS: We performed comprehensive immunohistochemical analysis of Glut-1 expression in 336 endometrial samples, including tumors, nontumor lesions, and normal tissues. RESULTS: Expression of Glut-1 was found in 87% of endometrioid carcinomas (160/184 cases), 100% of serous carcinomas (29/29 cases), 100% of clear cell carcinomas (17/17 cases), 50% of polyps with atypical hyperplasia (8/16 cases), 12.5% of polyps with non-atypical hyperplasia (3/24 cases), 77% of hyperplasias with atypias (10/13 cases), 9% of hyperplasias without atypias (1/11 cases), 87% of secretory endometrium samples (13/15 cases), and in none of the nonsecretory endometrium samples (0/27 cases). In endometrioid carcinomas, Glut-1 was expressed in a marked geographical pattern. In nontumor lesions, its expression was more common in atypical hyperplasia and polyps with atypical hyperplasia compared with polyps with non-atypical hyperplasia and hyperplasias without atypia ( P = .00032). CONCLUSION: Our study confirms the high expression of Glut-1 not only in endometrioid carcinomas but also in other carcinomas of endometrium including clear cell and serous types. Glut-1 expression can be used as a surrogate marker in differential diagnosis between hyperplasia with and without atypia. Because of common Glut-1 expression in malignant tumors, therapeutic strategies influencing this protein or its signaling pathways can be beneficial.
- MeSH
- Adenocarcinoma, Clear Cell diagnosis pathology MeSH
- Biopsy MeSH
- Diagnosis, Differential MeSH
- Adult MeSH
- Endometrium pathology MeSH
- Carcinoma, Endometrioid diagnosis pathology MeSH
- Endometrial Hyperplasia diagnosis pathology MeSH
- Immunohistochemistry MeSH
- Middle Aged MeSH
- Humans MeSH
- Young Adult MeSH
- Biomarkers, Tumor metabolism MeSH
- Endometrial Neoplasms diagnosis pathology MeSH
- Polyps diagnosis pathology MeSH
- Precancerous Conditions diagnosis pathology MeSH
- Glucose Transporter Type 1 metabolism MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Neoplasm Staging MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Young Adult MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Background: Endometriosis is a common gynecological disease characterized by the presence of endometrial tissue outside the uterus causing chronic inflammation, severe pain, and infertility. However, the innate immunity of gamma-delta (γδ) T lymphocytes in endometriosis has not been characterized. Women with endometriosis present numerous endocrine and immune dysfunctions and elevated risk for endometrial, ovarian, and breast cancers. The tyrosine kinase EphA2 is often overexpressed in cancer including endometrial carcinoma. Methods: We analyzed Vδ1 and Vδ2 γδ T cells in peripheral blood and paired peritoneal fluid samples in endometriosis patients (n = 19) and compared the counts with that of age- and sex-matched healthy donors (n = 33) using flow cytometry. Vδ1 and Vδ2 T cells isolated from healthy donors were used against KLE, RL-95, and Ishikawa endometrial tumor cells in 4 h flow cytometric cytotoxicity assays. The EphA2 blocking studies were performed using antibody, small-molecule inhibitor ALW-II-41-27, and the CRISPR/Cas9. Results: We determined Vδ1 T cells substantially reduced in patients' peripheral blood (p < 0.01) and peritoneal fluid (p < 0.001). No differences were found for circulating Vδ2 T cells compared with peritoneal fluid samples. We observed inherent cytotoxic reactivity of Vδ1 and Vδ2 γδ T lymphocytes against endometrial tumor cells. Importantly, we found reduced specific lysis of EphA2-positive cell lines KLE and RL-95 by Vδ1 T cells in the EphA2 antibody blocking studies and by the EphA2 inhibitor. Furthermore, Vδ1 T-cell-mediated killing was significantly decreased in RL-95 cell EPHA2 knockout. Finally, potent cytolytic activity exerted by Vδ1 T cells was significantly reduced in EPHA2 knockouts in renal A-498 and colon HT-29 carcinoma cell lines. Conclusions: We determined variable levels of Vδ1 and Vδ2 γδ T cells in endometriosis patients. We observed inherent cytotoxic reactivity of γδ T-cell subsets against endometrial cell lines. Specifically, we found that blocking of EphA2 expression resulted in significant inhibition of endometrial tumor killing mediated by Vδ1 γδ T cells. These results suggest that EphA2 is involved in tumor cell lysis and contributes to susceptibility to Vδ1 γδ T cells cytotoxic reactivity.
- MeSH
- Adult MeSH
- Endometriosis immunology metabolism MeSH
- Intraepithelial Lymphocytes immunology MeSH
- Middle Aged MeSH
- Humans MeSH
- Cell Line, Tumor MeSH
- Endometrial Neoplasms immunology metabolism MeSH
- Receptor, EphA2 metabolism MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
BACKGROUND: In the primary treatment of apparent uterine-confined endometrial carcinoma, pelvic ± para-aortic lymphadenectomy has been considered the standard of care. Although some retrospective data suggest that the sentinel lymph node algorithm without complete lymphadenectomy can be used without jeopardizing oncologic outcome, prospective data are lacking. PRIMARY OBJECTIVES: To assess the 36 month incidence of pelvic/non-vaginal recurrence in women with pathologically confirmed stage I intermediate-risk endometrioid endometrial carcinoma who have bilateral negative pelvic sentinel lymph nodes. STUDY HYPOTHESIS: We hypothesize that patients with stage I, intermediate-risk endometrioid endometrial carcinoma who have bilateral negative pelvic sentinel lymph nodes will demonstrate a pelvic/non-vaginal recurrence rate comparable to historical estimate of stage I, intermediate-risk endometrioid endometrial carcinoma patients (estimated 2.5%). TRIAL DESIGN: This prospective multicenter single-arm observational study will follow women with stage I, intermediate risk endometrioid endometrial adenocarcinoma who have undergone successful hysterectomy, bilateral salpingo-oophorectomy, and bilateral sentinel lymph node biopsies, for recurrence. All patients will undergo lymphatic mapping using indocynanine green and will either receive no adjuvant treatment or vaginal brachytherapy only. Patients will be followed for 36 months. MAJOR INCLUSION/EXCLUSION CRITERIA: Patients will be enrolled in the study cohort if all the following criteria are met: (i) at time of surgery: hysterectomy with bilateral adnexectomy, and successful bilateral pelvic sentinel lymph node mapping; (ii) on final pathology: pathologic stage I, intermediate-risk endometrioid endometrial carcinoma (grade 1 or grade 2 with ≥50% myometrial invasion, or grade 3 with <50% myometrial invasion), negative pelvic peritoneal cytology, and bilateral sentinel lymph nodes negative for malignancy; (iii) recommended adjuvant treatment: vaginal brachytherapy or no adjuvant treatment. PRIMARY ENDPOINT: Incidence of pelvic/non-vaginal recurrence at 36 months. SAMPLE SIZE: 182 patients for study cohort ESTIMATED DATES FOR COMPLETING ACCRUAL AND PRESENTING RESULTS: Accrual will be completed in 2023 with results reported in 2026. TRIAL REGISTRATION: NCT04291612.
- MeSH
- Adenocarcinoma pathology MeSH
- Sentinel Lymph Node Biopsy methods MeSH
- Humans MeSH
- Neoplasm Recurrence, Local diagnosis MeSH
- Lymphatic Metastasis pathology MeSH
- Endometrial Neoplasms pathology MeSH
- Prospective Studies MeSH
- Sentinel Lymph Node pathology MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
- Observational Study MeSH
- Research Support, N.I.H., Extramural MeSH
A specific form of endometrial cancer (EC) can develop in breast cancer patients previously treated with tamoxifen (ET), an antagonist of estrogen receptor (ER) that inhibits proliferation of ER positive breast cancer. ET tumors have a different phenotype than endometrial tumors, which typically develop de novo without previous exposure to tamoxifen (EN). Here we aimed to identify specific protein markers that could serve as specific molecular targets in either phenotype. A set of total 45 formalin-fixed paraffin-embedded (FFPE) endometrial tumor tissues and adjacent myometrium tissue samples were analyzed using LC-MS/MS in SWATH-MS mode. We found that calcyphosin (CAPS) levels were elevated in EN tumors compared to ET tumors. The higher CAPS level in EC tissue invading to myometrium supports its relationship to EC aggressiveness. Further, stathmin (STMN1) levels were found significantly elevated in ET versus EN tumors and significantly associated with patient survival. This finding connects elevated levels of this cell cycle regulating, proliferation-associated protein with tamoxifen exposure. In summary, using SWATH-MS we show that CAPS and STMN1 should be recognized as clinicopathologically different EC markers of which STMN1 is specifically connected with a previous tamoxifen exposition.
- MeSH
- Chromatography, Liquid MeSH
- Humans MeSH
- Endometrial Neoplasms * drug therapy MeSH
- Breast Neoplasms * MeSH
- Stathmin genetics MeSH
- Tamoxifen adverse effects MeSH
- Tandem Mass Spectrometry MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
