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BACKGROUND Endometrial cancers (EC) are a heterogeneous group of malignant neoplasms differing in etiology, clinical-pathological features and prognosis. OBJECTIVES To determine the differences between the expression of selected molecular factors and find connections between them in order to isolate possible biomarkers influencing treatment options. MATERIAL AND METHODS The investigated data involved archival histological preparations obtained from uterine EC samples taken from 137 patients, treated surgically between 2007 and 2014. The immunohistochemical Dako EnVisionTM Flex+ method was applied. RESULTS The expression of ERβ, MLH1 and BRCA1 was lower in ECI than in ECII patients. The ERα expression was higher in early Federation internationale de gynecologie et d'obstetrique (FIGO) (IA) stages than in advanced (IB-IV) stages, while ERβ expression was significantly higher in advanced stages compared to stage IA and increased with grading. The BRCA1 expression also increased with grading. In both type I and type II EC patients, ERα expression correlated with MYH9 and BRCA1, while ERβ expression correlated with BAP expression. High expression of BRCA1 correlated with several proteins: BAP, MYH9 and FAK. High BAP expression also correlated with high MYH9 expression. A correlation in the expression of these proteins was also demonstrated in the group consisting only of patients with ECI. A significant correlation was found between BAP expression and MYH9 among patients diagnosed with ECI. In the ECII group, no correlation was found between the tested proteins. CONCLUSIONS The ECI and ECII patients differed in the studied molecular factors, mainly in terms of ER and BRCA1 expression. Changes in BRCA1 expression were linked to alterations in BAP expression, but were also associated with the proteins MYH9 and FAK.

The aim of the study was to assess the activity of fucoidan on the uterine sarcomas (MES-SA and ESS-1) and carcinosarcoma cell lines (SK-UT-1 and SK-UT-1B) and its toxicity on the human skin fibroblasts (HSF). Two uterine sarcomas and two carcinosarcoma cell lines were examined, as a control HSF were used. Cell viability was assessed with MTT test, apoptosis with caspase-3 activity and cell cycle by assessment of DNA synthesis. Fucoidan significantly decreases cell viability in SK-UT-1, SK-UT-1B, and ESS-1 cell lines, such effect was not observed in MES-SA. Fucoidan was not substantially affecting proliferation among normal cells. The tested agent induced apoptosis in all cell cultures used in the experiment. Fucoidan affects cell cycle of all tested cell lines except MES-SA by increasing percentage of cells in G0/sub-G1/G1 phase. Fucoidan do not only affect proliferation but induces apoptosis in selected uterine sarcoma and carcinosarcoma cell lines, so it has potential to be used as cytotoxic agent. Fucoidan seems to be promising anti-cancer agent for endometrial stromal sarcoma and carcinosarcoma. Electronic supplementary material The online version of this article (10.1007/s00005-019-00534-9) contains supplementary material, which is available to authorized users.

Recent evidence suggests that immunological aspects play a pivotal role in this disorder. Toll-like receptor 2 (TLR2) is crucial in recognizing microbial infections and mediating innate immune response. The objective of our study was to rate with flow cytometry the levels of several subsets of dendritic cells, monocytes, and basic peripheral blood lymphocytes expressing TLR2, aiming at the determination of a possible correlation between the expression of TLR2 and the clinical outcomes of endometriosis in 40 patients and 40 age-matched healthy women. Our study showed the importance of TLR2 expression, mainly on myeloid dendritic cells (mDCs) and B cells in patients with endometriosis. Both mDCs BDCA1+CD19-TLR2+ and B lymphocytes CD19+TLR-2+ proved useful in the differentiation of affected individuals with stages 3&ndash, 4 of the disease (area under the receiver operating characteristic curve /AUC/ = 0.96, p &lt, 0.0001 for mDCs, AUC = 0.78, p = 0.0001 for B lymphocytes), and those presenting adhesion (AUC = 0.92, p &lt, AUC = 0.82, p &lt, 0.0001 for B lymphocytes) or infertility (AUC = 0.83, p &lt, AUC = 0.73, p = 0.006 for B lymphocytes). Our findings suggest that the levels of TLR2-expressing cells, particularly mDCs and B lymphocytes, may be an effective biomarker of endometriosis, because the disease currently lacks clinically useful noninvasive biomarkers enabling early and cost-effective diagnosis.

Previous studies have shown clinical relevance of programmed death-ligand 1 (PD-L1) and soluble PD-L1 (sPD-L1) in human cancers. However, still contradictory results exist. Our aim was evaluation of PD-L1-expressing monocytic myeloid-derived suppressor cells (M-MDSCs), monocytes/macrophages (MO/MA), tumour cells (TC) and immune/inflammatory cells (IC) as well as investigation of the sPD-L1 in ovarian cancer (OC) patients.The group of 74 pretreatment women were enrollment to the study. The expression of PD-L1 on M-MDSCS and MO/MA was assessed by flow cytometry. The profile of sPD-L1 was examined with ELISA. The expression of PD-L1 in mononuclear cells (MCs) was analyzed using real time PCR. PD-L1 immunohistochemical analysis was prepared on TC and IC. An in silico validation of prognostic significance of PD-L1 mRNA expression was performed based microarray datasets.OC patients had significantly higher frequency of MO/MA versus M-MDSC in the blood, ascites and tumour (each p 0.0001). In contrast, PD-L1 expression was higher on M-MDSCs versus MO/MA in the blood and ascites (each p 0.0001), but not in the tumour (p 0.05). Significantly higher accumulation of blood-circulating M-MDSC, MO/MA, PD-L1Although PD-L1 may not be a prognostic factor for OC, our study demonstrated impaired immunity manifested by up-regulation of PD-L1/sPD-L1. Furthermore, there was a positive association between PD-L1

Introduction: mTOR inhibitors are anticancer agents affecting mTOR/AKT/PI3K pathway that is one of the most important in human cancer cells. Hyperactivation of mTOR/AKT/PI3K and overexpression of this pathway members are frequently reported in uterine sarcoma and carcinosarcoma. Present study is aimed to assess the activity of the two mTOR inhibitors (rapamycin - RAP and sapanisertib - MLN) as a single agent and combined with gemcitabine (GEM, one of substances commonly used in systemic anticancer treatment) in uterine sarcoma and carcinosarcoma in vitro models. Material and methods: SK-UT-1 and SK-UT1-B (uterine carcinosarcoma), MES-SA (leiomyosarcoma) and ESS-1 (endometrial stromal sarcoma) cell lines were used. An MTT assay was performed to examine the cytotoxicity of RAP, MLN and mixtures: RAP+MLN, RAP+GEM, MLN+GEM against these cells. The interactions between tested compounds were assessed in isobolographic analysis. Results and conclusions: Carcinosarcoma cell lines (both SK-UT-1 and SK-UT-1B) do not respond to RAP and respond relatively weakly to MLN treatment. Additive and supraadditive effects were noted for combined treatment with GEM and MLN. Endometrial stromal sarcoma cell line (ESS-1) occured to be sensitive to both RAP and MLN, but the response was stronger for MLN. Additive effect of all tested drug combinations was observed for ESS-1. Leiomyosarcoma cell line (MES-SA) was found sensitive to both mTOR inhibitors. Additive effects in combinations of GEM, RAP and MLN were observed, what makes them promising for future preclinical and clinical trials. Additivity with slight tendency towards antagonism between GEM and MLN observed in MES-SA cell line is unexpected finding and might prompt the mechanistic research aimed to explain this phenomenon.

Vitamin D

One of the many factors involved in the development of uterine fibroids is vitamin D deficiency. One aspect of this deficiency is decreased serum concentration of calcidiol-25(OH)D, a metabolite of D3 vitamin. The active form of vitamin D3, which arises after numerous enzymatic reactions, is calcitriol-1,25(OH)2D3; this compound is transported to various body tissues. Vitamin D possesses extra-genomic effects due to its influence on various signaling pathways, i.e., through activating tyrosine kinases and by genomic effects via binding to a specific nuclear receptor, vitamin D receptor (VDR). The vitamin D/VDR complex regulates the expression of genes and is involved in the pathogenesis of fibroids. Numerous studies have shown that vitamin D supplementation reduces fibroid size. It has also been shown that the expression of VDR in myoma tissue is significantly lower than in the uterine muscle tissue at the tumor periphery. However, the expression of VDR in non-myoma uterine muscle has not previously been investigated. Our VDR expression studies were performed immunohistochemically with tissue microarrays (TMA) in three tissue groups: 98 uterine myoma tissues, 98 uterine tissues (tumor margin), and 12 tissues of normal uterine muscle (i.e., without fibroids). A statistical analysis showed significantly lower VDR expression in uterine muscle at the periphery of the fibroid than in healthy uterine muscle. Lower expression of VDR at the periphery of the myoma compared to that in normal uterine muscle may indicate potential for new myomas. This observation and the described reduction in the size of fibroids after vitamin D supplementation supports the hypothesis of causal development of uterine fibroids and may be useful for the prevention of re-development in the event of their excision from the uterus.

Introduction Data on the possible role of peritoneal fluid free radical-mediated oxidative damage in the pathogenesis of endometriosis still remains inconsistent. The aim of the study was to determine iron metabolism markers and their influence on oxidative stress arameters in the peritoneal fluid of women with endometriosis. Material and methods 110 women with endometriosis and 119 patients with benign ovarian cysts were included in the study. All visible peritoneal fluid was aspirated during laparoscopy from the anterior and posterior cul-de-sacs. under direct vision to avoid blood contamination. Haemoglobin, iron, total oxidative status, and total antioxidant status were measured using standard colourimetric kits. Results Haemoglobin, iron levels, as well as total oxidative status values were significantly higher, whereas total antioxidant status values were significantly lower in the peritoneal fluid of patients with endometriosis, in comparison to the reference groups. No differences were observed in peritoneal fluid concentrations of all parameters measured in relation to the phase of the menstrual cycle. Conclusions Peritoneal fluid of women with endometriosis is characterized by disrupted iron metabolism. This is most likely related to an increased number of erythrocytes in the peritoneal cavity of endometriotic women, which leads to a higher concentration of haemoglobin in this environment. Impaired iron homeostasis may have a significant influence on the pathophysiology of peritoneal endometriosis by the direct impact of haemoglobin derivatives and/or formation of the pro-inflammatory and pro-oxidative environment. Peritoneal cavity oxidative stress occurs predominantly in women in advanced stages of the disease.

PURPOSE The aim of this study was to evaluate the influence of ovarian cancer cell lysates isolated from type I or type II ovarian cancer (OC) on the phenotype of monocyte-derived dendritic cells (Mo-DCs) and the cytokine profile. We also determined whether the Mo-DCs and tumor microenvironment, reflected by peritoneal fluid (PF) from type I or II ovarian cancer, could promote regulatory T cell (Tregs) differentiation from naive CD4+ lymphocytes in vitro. RESULTS Our results show a significant role of the ovarian cancer microenvironment reflected by PF from type I or II OC in the inhibition of the DC differentiation process. Interestingly, the percentage of cells co-expressing CD45 and CD14 antigens in the cultures stimulated with PF from both type I and type II OC was higher than in the control. Furthermore, the percentage of cells expressing CD1a, i.e., a marker of immature DCs, was significantly reduced in the cultures stimulated with PF from type I and type II OC. The results obtained show that ovarian cancer type II lysates induce differentiation of monocytes into macrophage-like cells with a CD1a+/HLA-DR+/CD83− phenotype and significantly higher CD86/HLA-DR expression. We show that ovarian cancer type II Mo-DCs are able to prevent an immune response by release of IL-10, whereas OC type I Mo-DCs can promote the generation of Tregs. CONCLUSIONS We demonstrate that each type of ovarian cancer can induce a unique phenotype of DCs and differentiation of Tregs, both associated with immune-suppressive function, which may be an obstacle while developing effective anticancer dendritic cell vaccination.

Myeloid-derived suppressor cells (MDSCs) expansion is a hallmark of cancer. Three major MDSC subsets defined as monocytic (M)-MDSCs, polymorphonuclear (PMN)-MDSCs and early stage (e)MDSCs can be revealed in human diseases. However, the clinical relevance and immunosupressive pattern of these cells in epithelial ovarian cancer (EOC) are unknown. Therefore, we performed a comprehensive analysis of each MDSC subset and immunosupressive factors in the peripheral blood (PB), peritoneal fluid (PF), and the tumor tissue (TT) samples from EOC and integrated this data with the patients' clinicopathological characteristic. MDSCs were analyzed using multicolor flow cytometry. Immunosuppressive factors analysis was performed with ELISA and qRT-PCR. The level of M-MDSCs in the PB/PF/TT of EOC was significantly higher than in healthy donors (HD); frequency of PMN-MDSCs was significantly greater in the TT than in the PB/PF and HD; while the level of eMDSCs was greater in the PB compared with the PF and HD. Elevated abundance of tumor-infiltrating M-MDSCs was associated with advanced stage and high grade of EOC. An analysis of immunosuppressive pattern showed significantly increased blood-circulating ARG/IDO/IL-10-expressing M- and PMN-MDSCs in the EOC patients compared with HD and differences in the accumulation of these subsets in the three tumor immune microenvironments (TIME). This accumulation was positively correlated with levels of TGF-β and ARG1 in the plasma and PF. Low level of blood-circulating and tumor-infiltrating M-MDSCs, but neither PMN-MDSCs nor eMDSCs was strongly associated with prolonged survival in ovarian cancer patients. Our results highlight M-MDSCs as the subset with potential the highest clinical significance.

Purpose Cerebral metastases develop in 10–30% of patients with breast cancer (BC) and in around 3.3 to 4% of patients with ovarian cancer (OC). The aim of the multicenter study is to investigate the correlation between the expression of estrogen alpha receptors (ERα), progesterone receptors (PR), human epidermal growth factor receptor 2 (HER2), stromal cell-derived factor 1 (SDF1) and its receptor C-X-C chemokine receptor type 4 (CXCR4), breast cancer metastasis suppressor 1 (BRMS1), astrocyte elevated gene 1 (AEG1), depending on the status of BRCA1 protein, in patients suffering from OC and BC with brain metastases. Patients and methods The analysis included 51 patients: 29 with BC and 22 with OC, in whom brain metastases were disclosed. Results In most patients (65.5% of BC patients and 68.2% of patients with OC tumors) BRCA1 protein loss was found. No correlation was disclosed between the levels of ERα, PR receptors, HER2, SDF1, CXCR4, AEG1, BRMS1 and BRCA1 status, patient age, stage of disease advancement, grade of histological maturity of the cells, presence of metastases to lymph nodes. A statistically significant correlation was disclosed between the negative expression of PR receptors and a high expression of CXCR4 in patients with BC. High values of the AEG1 protein (linked to metastases) were detected alongside a high expression of BRMS1 (a suppressor of metastases). Conclusions Patients with BC and OC and brain metastases have a frequent loss of BRCA1 expression. The role of ERα, PR, HER2, SDF1, CXCR4, AEG1, BRMS1 in metastatic process needs further studies.

Objectives: The objective of the study was to retrospectively evaluate the density of vessels exhibiting positive glycoprotein CD34 expression in the uterine leiomyosarcoma tissues and their correlation with the age of patients at the time of tumor diagnosis. Material and methods: The archival paraffin blocks with the cancer tissues collected from 50 patients suffering from uterine leiomyosarcoma were used together with their clinical and demographic data. The immunohistochemical peroxidase-de­pendent methods were used to detect microvessels with positive CD34 expression. The glycoprotein CD34 expression was evaluated as a density of microvessel showing the positive immunohistochemical reaction (MVD CD34 ). Results: The negative, statistically significant correlation between the age of patients (at the moment diagnosis) and the MVDCD34+ (R = –0.289, p = 0.042) was found. Conclusions: The study’s findings may suggest that the tissues of younger people constitute a permissive environment for pro-angiogenic factors.

BACKGROUND Endometrial carcinomas (EC) differ in etiology, clinical course and prognosis. OBJECTIVES This multi-center study aimed at a closer recognition of molecular factors linked to heterogeneity of EC by evaluating estrogen and progesterone receptors, proteins dependent on MMR genes, proteins linked to poor prognosis and metastases, and mutations in BRCA1. MATERIAL AND METHODS Using sections of paraffin-embedded preparations, in 115 patients with EC type I and 31 with EC type II, expression of ERα, ERβ1, PR, MLH1, and MSH2 proteins, as well as ARID1A, c-MET and BRCA1, was estimated by immunohistochemistry using specific antibodies. RESULTS Expression of ERβ1 was augmented in EC type II, in poorly differentiated cancers and with growing clinical advancement. An augmented expression of ERα was noted in well-differentiated EC and at lower clinical stage. An increased expression of PR and decreased of MLH1 were detected in type I EC. The expression of ARID1A and c-MET proteins showed no differences between the types of EC, stages of clinical advancement or grading. In 51.6% patients with type II EC, a loss of BRCA1 expression was disclosed; in this group of cancers a decreased expression of ERα was noted. CONCLUSIONS An augmented expression of ERβ1 was linked to type II EC. A higher expression of ERα in EC cancers was associated with a lower histopathological grade. A decreased expression of MLH1 protein was estimated in EC type I. Type II EC may be connected to BRCA1 mutation.

Ovarian cancer is a serious diagnostic and clinical issue. It belongs to the group of cancers with the highest mortality rate, that is why new, effective methods of therapy have been sought after. In recent years, researchers have been paying attention to the use of immunothetapy in the treatment of ovarian cancer. Currently, the numer of studies with the use of PD-1/PD-L1 pathway inhibitors is increasing. It has been reported that PD-1 receptor and its ligand are expressed on tumor cells and immunology system cells in patients with ovarian cancer. Increased expression of PD-1/PD-L1 is one of the inhibition mechanisms of the anti-tumor response by induction of peripheral tolerance. That seems why blocking PD-1/PD-L1 may be so important. A significant role in activation of programmed death cell-1 is attributed to tumor microenvironment (TME). In this review we have described the meaning of PD-1/PD-L1 pathway in ovarian cancer pathogenesis and current results of clinical trials using PD-1/PD-L1 inhibitors. Numerous clinical trials are focused on the effectiveness of immunotherapies as both monotherapy and combination therapy. The promising results of initial research phases are the basis for taking action on a larger scale. Perhaps this will allow in the future to use inhibitors of the PD-1/PD-L1 pathway in the treatment of ovarian cancer.

Introduction and objective. The objective of the study was to present preliminary results of a pilot study concerning the level of knowledge of gynaecological prevention, conducted in teenagers referred by court rulings to a juvenile attendance centre. Materials and method. The instrument was an anonymous survey questionnaire completed by participants in health prevention classes in late 2010 and early 2011. The studied group consisted of teenagers aged 15–17 years (mean age: 15.72 years, median = 16 years, SD = 0.679) who were under probation officers’ supervision in the Zamość region of south-eastern Poland. The sample size was 101 persons – 51 boys (50.50%), 50 girls (49.5%). Results. According to the respondents, the most important reasons for seeing a gynaecologist were: menstrual disorders (70.30%), suspicion of pregnancy (63.37%) and pain or burning sensations while urinating (58.42%). The following were regarded as prevalent cancers in women: cancers of the breast (99.01%), cervix (89.1%), and ovaries (62.38%). Over 92% of subjects stated that it was possible to protect oneself from cervical cancer, but only 41.5% of respondents indicated the correct definition of the term ‘cytology’. Statistical analysis focused on differences between genders. A higher self-assessment of mental health was shown in boys. Conclusions. Teenagers of similar background may find it more difficult to gain access to knowledge about health prophylaxis, including gynaecological prevention. Efforts should be intensified in order to ‘equalize health opportunities’ through appropriate preparation of teaching curricula (including health education and philosophy of medicine).

Uterine leiomyosarcomas (LMS) are rare tumors typically presenting rapid growth and unfavorable outcome. Nowadays the results of uterine LMS treatment do not meet expectations. Angiogenesis is one of processes investigated to be target for future treatment. The aim of the research was to assess microvessels density (MVD) in tumor samples collected from 50 patients with histological confirmed uterine leiomyosarcoma and to investigate statistical relations between MVD, patients survival, and FIGO stage of tumor. The assessment was carried out using immunohistochemistry methods with anti-CD34 antibody. No significant difference in MVD between FIGO stages was observed. Furthermore, contrary to many other malignancies, we found no significant relation between MVD and patients overall and 2-year survival. Results obtained in the study suggest that processes on vascular mimicry and mesenchymal to epithelial transition (MET) may play important role in development of LMS. No statistical relation between MVD and survival leads to conclusion that not only angiogenesis but other mechanisms as well should be taken into consideration in planning future research.

Objectives: The role of angiogenesis in leiomyosarcomas still remains unclear. The aim of this study was to evaluate the NRP1 expression in the leiomyosarcoma tissues and to find the relations between its expression and the clinical features. Material and methods: The study group consisted of 50 patients with diagnosis of the uterine leiomyosarcoma. Clinical and follow up data were collected. Using immunohistochemical methods the expression of NRP1 was detected. Results: The lack of NRP1 expression was found in 14 cases, positive (weak or moderate) expression was noted in 36 cases. The significantly higher expression of NRP1 was observed in more severe clinical stages in comparison to lower stages of the disease. The significantly shorter survival of patients with the positive expression of NRP1 in leiomyosarcoma was observed. Conclusions: The expression of NRP1 is associated with clinical advancement and worse prognosis in uterine LMS. Neuropilin 1 can be widely used as a postoperative survival predictor for the patients suffering from uterine LMS.

Ovarian cancer is a malignancy of high mortality rates. In respect of the number of deaths caused by cancers it occupies the fourth place among women in Poland. Recent studies are focusing on the role of immune system in ovarian cancer pathogenesis. It has been reported that immune response against ovarian cancer cells may be inhibited by a number of immunosuppressive mechanisms active in cancer microenvironment. It causes difficulties in recognizing and destroying cancer cells by immune system which leads to the development of immune tolerance and is associated with a low efficacy of standard therapeutic strategies. In the presented paper we have described selected, new immunosuppressive mechanisms in ovarian cancer patients. They may be a novel, additional and relevant criterion that should be considered whilst developing new therapeutic strategies. Possibly, modulation of immunosuppressive mechanisms could contribute to modifying standard therapies and in consequence improve treatment outcome in ovarian cancer patients.

The low incidence of uterine sarcomas requires many issues associated with its biology and clinical course to be followed with more research. Unsatisfactory surgical outcomes and a high risk of cancer dissemination make it worthwhile to consider the feasibility of supplementary systemic treatment. The currently employed chemo- and hormonal therapy is characterised by low efficacy. There is some hope in reports on targeted treatment. However, a comprehensive assessment of the efficacy of this kind of therapy is restricted by a small number of patients using it. Moreover, clinical studies using targeted therapies involve patients with a highly advanced disease, and the therapeutic results are assessed mainly via analysis of progression-free survival but not the clinical response.

Objective To evaluate 8-hydroxy-2-deoxyguanosine (8-OHdG) and 8-isoprostane levels in the peritoneal fluid (PF) of women with endometriosis. Study design One hundred and ten women with laparoscopically and histopathologically confirmed endometriosis and, as reference groups, 119 patients with simple serous (n = 78) and dermoid (n = 41) ovarian cysts were studied. Peritoneal fluid 8-OHdG and 8-isoprostane concentrations were evaluated by enzyme-linked immunosorbent assays. Results 8-OHdG and 8-isoprostane levels in peritoneal fluid were significantly higher in patients with endometriosis compared with the reference groups. Higher PF 8-OHdG and 8-isoprostane concentrations were observed in patients with advanced stages of endometriosis. A statistically significant positive correlation was found between 8-OHdG and 8-isoprostane levels in peritoneal fluid. Conclusion Endometriosis induces greater oxidative stress and frequent DNA mutations in peritoneal fluid than nonendometriotic ovarian cysts. The most severe oxidative stress occurs in the peritoneal cavity of women with more advanced stages of the disease.

The majority of adult women with endometriosis report that their symptoms started in adolescence. Early diagnosis and optimal treatment prevent disease progression and mitigate long-term morbidities, such as infertility and chronic pelvic pain.

Cyclin A is a cell-cycle regulatory gene and its overexpression promotes tumor cell growth. Y-Box-binding protein 1 (YB1) is a transcription/translation factor involved in tumor growth, invasion, and drug resistance. We investigated whether an association exists between protein products of these genes in epithelial ovarian cancer (EOC) specimens and clinicopathological parameters, patient response and EOC sensitivity to platinum-based first-line chemotherapy.Cyclin A and YB1 expression were analyzed by immunohistochemistry in 54 human primary EOC tissues. Immunolabeling of both proteins was graded according to their staining intensity (scale 0-3) and the proportion of immunostained cancer cells (scale 0-4) to obtain a staining index (SI; value=0-12).Significantly higher cyclin A immunostaining (SI≥4) in EOC specimens was discovered in patients with advanced (International Federation of Gynaecology and Obstetrics (FIGO) III and IV, p=0.003), poorly differentiated (G3, p0.001) tumors, and tumors of those with residual disease1 cm (p=0.001). YB1 immunostaining was significantly higher in EOCs from patients with suboptimal debulking (p=0.025). Over-expression of cyclin A (SI≥9) in EOCs was significantly linked with poorer patient response (p=0.001) and higher resistance of tumors to platinum-based first-line chemotherapy (p=0.007), while immunolabeling of YB1 in EOCs was not significantly associated with either of these variables (p0.05). Cyclin A expression was significantly and positively correlated with that of YB1 (R=0.588, p0.001).Increased cyclin A expression in EOC is related to a more aggressive tumor behavior and predicts the response of patients to first-line platinum-based chemotherapy.

Objectives: Most investigators agree that endometriosis is associated with a state of subclinical, non-infectious peritoneal inflammation. The objective of the study was to assess concentrations of two markers of the acute inflammatory phase proteins, haptoglobin and ceruloplasmin, in peritoneal fluid of endometriotic women. Material and methods: 229 women who underwent diagnostic or therapeutic laparoscopy were included in the study. Minimal, mild, moderate and severe endometriosis according to ASRM was confirmed in 119 women (study groups), whereas 110 patients suffered from simple serous or dermoid ovarian cysts (reference groups). Haptoglobin and ceruloplasmin concentrations in the peritoneal fluid samples aspirated during laparoscopy were measured using commercially available radial immunodiffusion kits. Results: The concentration of haptoglobin in the peritoneal fluid of women with endometriosis was significantly higher as compared to patients with serous and dermoid ovarian cysts. Significantly higher haptoglobin level was observed in patients with severe and moderate endometriosis as compared to women from both reference groups. No significant difference in the peritoneal fluid ceruloplasmin levels was found between patients with endometriosis and women from reference groups. However, it was noted that ceruloplasmin levels are higher in the subgroup of patients with severe endometriosis as compared to both reference groups and women with mild disease. Conclusions: Our results support the hypothesis that endometriosis is associated with subclinical inflammation within the peritoneal cavity. It may be speculated that pro-inflammatory stimuli strong enough to cause an increase in acute inflammatory phase proteins peritoneal fluid concentrations are observed only in the advanced stages of the disease.

The etiopathogenesis of endometriosis still remains unknown. Recent data provide new valuable information concerning the role of oxidative stress in the pathophysiology of the disease. It has been proved that levels of different lipid peroxidation end products are increased in both peritoneal fluid (PF) and serum of endometriotic patients. We assessed the concentration of oxidized low-density lipoproteins (oxLDL) in PF of 110 women with different stages of endometriosis and 119 women with serous () or dermoid () ovarian cysts, as the reference groups. PF oxLDL levels were evaluated by ELISA. We found that concentrations of oxLDL in PF of endometriotic women were significantly higher compared to women with serous but not dermoid ovarian cysts. Interestingly, by analyzing concentrations of oxLDL in women with different stages of the disease, it was noted that they are significantly higher only in the subgroup of patients with stage IV endometriosis as compared to women with ovarian serous cysts. In case of minimal, mild, and moderate disease, PF oxLDL levels were similar to those noted in reference groups. Our results indicate that disrupted oxidative status in the peritoneal cavity of women with endometriosis may play a role in the pathogenesis of advanced stages of the disease.

The process of T cells activation is necessary for the performance of the defense functions. Successful activation depends on direct lymphocyte - antigen-presenting cell contact and signal transmission to the lymphocyte. Activated T cells exhibit surface expression of molecules such as CD69, CD25 and HLA-DR. The effect of cell activation is a cascade of molecular events leading to proliferation and clonal expansion of antigen-specific T cells.The aim of this study was to evaluate the phenotype and T cell activation markers: CD69, CD25 and HLA - DR in the peripheral blood and tumor tissue of ovarian cancer patients.The study group consisted of 26 patients operated due to ovarian cancer (FIGO Ilb - IV). Mononuclear immune cells were isolated from peripheral blood and ovarian cancer tissue. To obtain peripheral blood lymphocytes, blood was collected into heparinized tubes and diluted 1:1 with PBS, then layered on Gradisol L and centrifuged 20 minutes at 2800 rpm. Mononuclear cells were washed twice with PBS and labeled with monoclonal antibodies. A small piece of tumor tissue (about 1cm3) was fragmented with a surgical blade. Minced tissue was suspended in PBS and layered on Gradisol L for mononuclear cells isolation. To assess the phenotype and activation status of peripheral blood and tumor infiltrating lymphocytes, we used FACS Canto cytometer and monoclonal antibodies conjugated with fluorochromes: anti-CD3-FITC, anti-CD4-PE-Cy5, anti-CD8-APC, anti-CD25-PE, anti-CD69-PE-Cy7, anti-HLA-DR-PE-Cy7. Statistical analysis was performed using the Statistica 5.0 and Wilcoxon test.In all cases we detected T helper CD3+CD4+ and cytotoxic CD3+CD8+. T lymphocytes in both blood samples and tumor tissues. We observed no statistically significant difference in the percentage of CD3+ CD4+ cells among the mononuclear cells present in peripheral blood and tumor tissue. The percentage of CD3+CD8+ cytotoxic T lymphocytes was higher among mononuclear cells isolated from the tumor tissue. The percentage of CD3+ lymphocytes expressing the very early activation marker CD69 was significantly higher among tumor infiltrating lymphocytes compared with peripheral blood lymphocytes. Similarly the percentages of CD3+CD4+CD69+ T helper lymphocytes and CD3+CD8+CD69+ cytotoxic T lymphocytes were significantly higher on lymphocytes isolated from tumor tissue when compared to blood. The expression of an early activation marker - CD25 was significantly higher on the CD3+ and CD3+CD8+ peripheral blood lymphocytes compared to CD3+ and CD3+CD8+ tumor infiltrating lymphocytes. There were no statistically important differences between the percentages of, isolated from blood and tissue, CD3+CD4+ T helper lymphocytes. The expression of the late activation marker - HLA-DR was significantly higher on CD3+ lymphocytes isolated from tumor tissue compared with peripheral blood. Similarly the percentages of CD3+CD4+ lymphocytes and CD3+CD8+ cytotoxic T cells expressing HLA-DR were significantly higher among tumor infiltrating lymphocytes when compared to peripheral blood ones.T cells obtained from ovarian cancer tissues are activated cells. The state of T cell activation may be the result of direct contact of these cells with tumor antigens. The low expression of CD25 may suggest abnormal clonal expansion of antigen-specific lymphocytes.

Knowledge of the role of interleukin 17 (IL-17) has led to the identification of new subpopulation of T helper lymphocytes--Th17 and T cytotoxic lymphocytes secreting IL-17 (Tc17). An increasing amount of attention is paid to their role in anti-tumor immunity.The aim of this study was to evaluate the percentage of peripheral blood, peritoneal fluid and cancer tissue CD4+ and CD8+ T lymphocytes producing IL-17 in patients with ovarian cancerForty patients operated due to advanced ovarian carcinoma and twenty-four patients with functional follicle ovarian cysts were recruited. Peripheral blood, peritoneal fluid and cancer tissue mononuclear cells from ovarian cancer patients were stimulated for 4 hours ex vivo with phorbol myristate acetate (PMA) (50 ng/ ml) and ionomycin (1 microg/ml). The percentage of CD4+ and CD8+ T cells producing IL-17 was measured using flow cytometry.CD4+ and CD8+ T lymphocytes producing IL-17 were detected in the peripheral blood, peritoneal fluid and cancer tissue of ovarian cancer patients. The percentage of CD4+ T cells producing IL-17 was higher in the peripheral blood, peritoneal fluid and cancer tissue when compared to CD8+/IL 17+ T cells. The percentage of CD4+/ IL-17+ was significantly higher in cancer tissue compared to T cells derived form peripheral blood. There was no difference in the percentage of CD4+/IL-17 + T cells between peripheral blood and peritoneal fluid and peritoneal fluid and cancer tissue of ovarian cancer patients. There was no difference in the percentage of CD8+/IL-17 + T lymphocytes in the peripheral blood, peritoneal fluid and cancer tissue in patients suffering from ovarian cancer.Increased percentage of CD4+/IL-17+ and CD8+/IL-17+ T cells in cancer tissue indicates that these cells are accumulated in ovarian cancer microenvironment.

Podoplanin is a transmembrane glycoprotein expressed in endothelial lymphatic cells. It was proven to be a predictive marker in a variety of cancers e.g. mesothelioma and head and neck squamous-cell carcinoma. Ovarian clear cell carcinoma (OCCC) is a rare and unique histopathologic subtype of epithelial ovarian cancer (EOC). The molecular basis of that phenomenon remains unknown.The aim of our study was to assess podoplanin expression on the protein level in OCCC.Immunohistochemistry was performed on paraffin-embedded tissues from 19 patients with diagnosed OCCC.Podoplanin expression was present (moderate or strong) in 52% of OCCC cases (10/19). Nine of eleven (81.2%) postmenopausal and one of eight (12.5%) premenopausal women were podoplanin positive. No differences in podoplanin expression were found in relation to clinical features of the tumorThe incidence of podoplanin expression is higher in ovarian clear cell adenocarcinoma in postmenopausal patients.

This study was undertaken to evaluate RANTES levels in the peritoneal fluid (PF) and plasma of patients with ovarian cancer (n=73), serous cystadenoma (n=32) or normal controls (n=9). RANTES levels were correlated to myeloid and lymphoid dendritic cells (DCs). RANTES levels were evaluated using the ELISA assay. DCs were quantified using flow cytometry. The PF and plasma RANTES concentrations were elevated in the ovarian cancer (OVC) patients when compared to the patients with benign tumor (the reference group). Plasma levels of RANTES were higher in OVC patients compared with the reference group and with the controls. There were no significant differences in the plasma RANTES levels based on tumor stage, grade or histology. Women with serous cystadenocarcinoma, clear cell carcinoma and endometrioid cystadenocarcinoma had significantly higher PF RANTES levels than patients with undifferentiated carcinoma. Women with clear cell carcinoma and patients with endometrioid cystadenocarcinoma had higher PF RANTES levels than women with mucinous cystadenocarcinoma. We concluded that RANTES production in the peritoneal cavities of OVC patients depends on the histological type of the tumor cells.

Objectives. The aim of our study was to generate dendritic cells (DCs) from peripheral blood monocytes (PBMC) of patients suffering from ovarian cancer. Materials and Methods. Immature DCs were generated from PBMC cultured in RPMI 1640 medium with 2% human serum albumin (HSA), supplemented with recombinant human (rh) granulocyte macrophage colony stimulating factor (GM-CSF) and rh interleukin (IL)-4. After 5 days of culture, DC maturation was induced by the addition of an ovarian cancer cell line (CAOV3) lysate and after 6 days of rh tumor necrosis factor (TNF)-α for a further 2 days. The phenotype of the generated cells was assessed by flow cytometry for the expressions of human leukocyte antigen (HLA)-DR, costimulatory molecules (e.g., CD86, CD80), CD83, CD1a, and CD14. PBMC cultured in 2% HSA without rhIL-4, rhGM-CSF, rh-TNF-α, or tumor cell lysate formed the control group. Results. The 2.41% (interquartile range, 1.51%-3.52%) of CD45 + /CD14 + cells in cultures with rhIL-4, rhGM-CSF, rhTNF-α and tumor cell lysate was significantly lower than in the control group (31.10%; interquartile range, 11.11%-64.06%). Cultures with rhIL-4, rhGM-CSF, rhTNF-α, and tumor cell lysate showed a higher percentage (19.96%; interquartile range, 9.30%-24.42%) of fully mature CD83 + /CD1a - /HLA-DR + DCs compared with control culture (6.02%; interquartile range, 3.01%-7.37%). There was no significant difference in the expression of the immature DC marker (CD1a) between the cultures. The expression of co-stimulatory markers (CD80, CD86, HLA-DR) was greater (24.29%; interquartile range, 18.68%-33.95%) on DCs from cultures with rhIL-4, rhGM-CSF, TNF-α, and tumor cell lysate versus controls (4.93%; interquartile range, 2.67%-9.09%). Conclusion. Our data demonstrated that immature and mature DCs can be generated from adherent human PBMC from ovarian cancer patients cultured with rhIL-4, rhGM-CSF, rhTNF-α, and tumor cell lysates.

Neuropilin (NRP) may be used as a marker of lymphangiogenesis in malignant tumors. Significant correlations of the expression of NRP with tumor progression and overall survival prognosis were found in several cancer types. However; its potential role in epithelial ovarian cancer (EOC) has not been clarified.The aim of the work was to study a possible correlation of neuropilin-1 (NRP-1) expression with selected clinical and histological features of EOC.The study included 53 women (aged 23 to 81, mean 56.6 +/- 14.4 yrs), 38 of which were postmenopausal (71.7%). Immunohistochemical staining with a specific anti-NRP-1 antibody was performed in representative tumor tissue samples of patients with EOC. Both, percentage of stained lymphatic cells and intensity of staining were assessed under 200x magnification. The results were correlated with the menopausal status, FIGO stage, histological type and histological grade of EOC.Histological examination revealed that there were 27 cases of serous cancers (50%), 15 cases of mucinous cancer (28.3%) and 11 endometrioid cancers (20.7%). In 41.5% (n = 22) cases of EOC no NRP-1 staining was found, a weak (+) or strong (++) staining were found in 13 (24.5%) and 18 tumors, respectively There were no significant differences between neuropilin-1 expression and both menopausal status of women and histological type of EOC. Except for stage II, clinical EOC patients' FIGO stage was not correlated with the lack of expression (38.8% for stage I, 71.4% for stage II and 35.7% for stage III).We believe that neuropilin-1 expression is probably not related to clinical and histological features of epithelial ovarian cancer

One of the most commonly assessed angiogenesis markers is microvessel density which is determined on the bases of specific endothelial antigen expression (CD34, CD105). Angiogenesis modulators include growth factors and their receptors (EGFR), proteases and their inhibitors, oncogenes and suppressor genes (p53).The aim of the study was to evaluate whether there are any differences in selected angiogenesis markers and modulators expressions in ovarian cancer patients with different menopause status.The study included 100 women, age 30-70, who underwent surgical treatment due to ovarian cancer. As far as their menopause status was concerned, the women were divided into three groups: pre-, peri-, and postmenopausal. Microvessel density was assessed on the basis of CD34 (MVD(CD34) and CD 105 (MVD(CD105) expression. Additionally, tumor tissue p53 protein and EGFR expression were investigated. Active EGFR form in blood serum samples of cancer patients was assessed before the surgery.Microvessel density, assessed on the basis of CD34 and CD105 expression, as well as p53 and EGFR expression were similar in all three groups of patients. Active EGFR serum concentration in women with ovarian cancer did not prove to be significantly different and did not depend on the menopause status.Intensity of the angiogenesis process does not depend on the menopausal status of women and is similar in pre-, peri- and postmenopausal patients.

Background: Evaluation of immature myeloid and lymphoid dendritic cells (DCs) in the peritoneal fluid (PF) and peripheral blood (PB) mononuclears of women with ovarian carcinoma (n 5 47) and benign ovarian tumors (n 5 37). Methods: Mononuclear cells were isolated from PF and PB, stained with monoclonal antibodies (mAbs) against DC antigens (anti-BDCA-1, anti-BDCA-2), and estimated using flow cytometry. Results: The percentage of PF myeloid DC (MDC) in mononuclears was significantly lower in patients with ovarian cancer in comparison to the group of nonmalignant ovarian tumors (0.65% and 6.95%). The percentage of PF lymphoid DCs (LDCs) was higher in patients with ovarian cancer than in the reference group (0.64% and 0.09%). The percentage of PB MDCs and LDCs did not differ significantly between studied groups. In women suffering from ovarian cancer the percentage of both MDCs and LDCs was higher in the PF than in the PB. In the reference group the percentage of MDCs was higher but that of LDCs was lower in the PF than in the PB. In women with ovarian cancer, PF MDCs/LDCs ratio was lower in comparison to patients with serous cystadenoma. In PB the ratio of MDCs to LDCs did not differ significantly between studied groups. Conclusions: We concluded that MDCs population may be affected by the presence of malignant disease. LDC subsets may have influence on the local immune response in the PF of women with malignant tumors of the ovary. q 2008 Clinical Cytometry Society Key terms: myeloid DCs; lymphoid DCs; peritoneal fluid; cystadenocarcinoma

Tension-free vaginal tape (TVT) has within the last decade become one of the most popular procedure for treatment of female urinary stress incontinence. It is still persistent the risk of complications peri- and postoperative in spite of high efficacy of this method. Investigators still have been looking for new variations of this method to diminish the risk of complications and to maintain the efficacy of the treatment. The aim of this study is the description of the pre pubic TVT method and the preliminary short time (3 month) results of treatment. The results of pre-pubic TVT treatment were compared with "classic" TVT method. The pre-pubic TVT technique was performed in 25 women, operated for stress incontinence. The average operation time was 19 min (10-30 min). During the operation in one of patients there was excessive bleeding from venous plexes of vagina during preparation of tissues and in 4 of them it was perforation of vaginal mucosa during introducing of needle with tape. All these complications were recognised and corrected during the surgery. The mean hospitalisation time was two days. Our 3-months observations allow to confirm that efficacy of pre-pubic TVT is similar to the "classic" method, but the frequency of complications is lower in the pre-pubic TVT. There were no postoperative complications and voiding dysfunctions.

Of the study was to evaluate utility of magnetic resonance (MR) in the assessment of the women who fulfilled inclusion criteria for uterine artery embolisation as a symptomatic leiomyoma treatment and outcome of procedures.MR examination was performed in 220 patients. Uterine artery embolizations were performed in 170 patients.The reasons for disqualification were: no correlation between symptoms and size an position of leiomyoma--61%, adenomyosis--20%. In MR imaging dominant leiomyoma volumes were diminished by 59.6% (p0.05) over a period of 3 months.MR is the method of choice in the assessment of women referred to uterine artery embolisation as a symptomatic leiomyoma treatment. MR imaging is effective and objective method of evaluation outcomes of uterine embolisation.

Three-dimensional sonography is a novel diagnostic method proposed to be an additional non-invasive tool in the assessment of ovarian tumors.To study diagnostic potential of 3D sonography and power Doppler angiography in the preoperative differentiation of adnexal masses.One hundred twenty-eight women with tumors thought to be of adnexal origin were examined preoperatively. Following morphological (papillae, septae, tumor size and volume) and color Doppler (PI, RI, Vmax and TAMX) assessment, 3D ultrasound of adnexal tumors according to Kurjak et al. (2000) was performed. Various scanners were used and included: ATL 5000 HDI (Phillips, USA) and Combison 530 and Voluson 730 (Kretztechnik, Austria) machines. Following variables were studied: inner wall structure, presence of papillae, thickening3 mm of septa as well as vascular branching pattern, number and localization of small blood vessels and the presence of vascular anastomoses.Twenty-one tumors were malignant (3 FIGO stage I) and 101 masses were benign. Power Doppler combined with 3D sonography predicted malignancy with a sensitivity of 92.6% (25 of 27 patients). Commonly used morphological and Doppler criteria produced lower sensitivity, the values being in range of 45% to 87.5%. Negative predictive value of 97.2% was the highest for 3D sonography (7 points in Kurjak's scale).Selective use of 3D ultrasound and power Doppler angiography could be used to better characterize adnexal tumors. Detailed 3D sonography may help to identify women who, if needed, may have less invasive surgical procedure such as laparoscopy or be referred to gynecological oncologist.

The aim of the study was the evaluation of the way of diagnostic and therapeutic management in case of neoplasm of the reproductive organ in pregnant women treated in the I Clinic of the Operative Gynaecology in Lublin.In the years 1990-2000 in the I Clinic of the Operative Gynaecology in Lublin, 3556 operations were performed. In 9 cases the operations were carried out because of the neoplasm of the reproductive organ diagnosed in the course of pregnancy. In 8 cases, pregnant women with tumors in the pelvis minor were operated on and in 6 cases benign ovary neoplasms were stated.In one patient, extraperitoneal tumor coming out from the region of the sacral bone was diagnosed. In one case the patient in the 16th week of pregnancy was operated on and she had 10 cm in diameter myoma enucleated. The myoma was localized on the anterior uterus wall. In patient in the 33rd week of pregnancy with the diagnosed cervical carcinoma of the 2nd degree, diagnosed on the basis of histopathological examination, Caesarean section and Meigs operation were performed. In all operated patients tocolysis was employed. In one case there was a miscarriage in the 16th week of pregnancy following the myoma enucleation. The postoperative course in the remaining women was uncomplicated and pregnancies ended with the delivery of healthy infant.The most common neoplasms occurring in pregnancy are simple ovarian cysts and uterine myomas. Among malignant changes sporadically ovarian and cervical carcinomas are met. Although tumors of the reproductive organ in pregnant women are quite rarely met, the diagnostic and therapeutic difficulties reveal the necessity of the though differential diagnosis and accurate establishment of indications for the operation.

To assess the prevalence of human papilloma virus (HPV) in esophageal squamous cell carcinoma (ESCC) in the south-eastern region of Poland.The study population consisted of 56 ESCC patients and 35 controls. The controls were patients referred to our department due to other non-esophageal and non-oncological disorders with no gross or microscopic esophageal pathology as confirmed by endoscopy and histopathology. In the ESCC patients, samples were taken from normal mucosa (56 mucosa samples) and from the tumor (56 tumor samples). Tissue samples from the controls were taken from normal mucosa of the middle esophagus (35 control samples). Quantitative determination of DNA was carried out using a spectrophotometric method. Genomic DNA was isolated using the QIAamp DNA Midi Kit. HPV infection was identified following PCR amplification of the HPV gene sequence, using primers MY09 and MY11 complementary to the genome sequence of at least 33 types of HPV. The sequencing results were computationally analyzed using the basic local alignment search tool database.In tumor samples, HPV DNA was identified in 28 of 56 patients (50%). High risk HPV phenotypes (16 or/and 18) were found in 5 of 56 patients (8.9%), low risk in 19 of 56 patients (33.9%) and other types of HPV (37, 81, 97, CP6108) in 4 of 56 patients (7.1%). In mucosa samples, HPV DNA was isolated in 21 of 56 patients (37.5%). High risk HPV DNA was confirmed in 3 of 56 patients (5.3%), low risk HPV DNA in 12 of 56 patients (21.4%), and other types of HPV in 6 of 56 patients (10.7%). In control samples, HPV DNA was identified in 4 of 35 patients (11.4%) with no high risk HPV. The occurrence of HPV in ESCC patients was significantly higher than in the controls [28 of 56 (50%) vs 4 of 35 (11.4%), P0.001]. In esophageal cancer patients, both in tumor and mucosa samples, the predominant HPV phenotypes were low risk HPV, isolated 4 times more frequently than high risk phenotypes [19 of 56 (33.9%) vs 5 of 56 (8.9%), P0.001]. A higher prevalence of HPV was identified in female patients (71.4% vs 46.9%). Accordingly, the high risk phenotypes were isolated more frequently in female patients and this difference reached statistical significance [3 of 7 (42.9%) vs 2 of 49 (4.1%), P0.05]. Of the pathological characteristics, only an infiltrative pattern of macroscopic tumor type significantly correlated with the presence of HPV DNA in ESCC samples [20 of 27 (74.1%) vs 8 of 29 (27.6%) for ulcerative or protruding macroscopic type, P0.05]. The occurrence of total HPV DNA and both HPV high or low risk phenotypes did not significantly differ with regard to particular grades of cellular differentiation, phases in depth of tumor infiltration, grades of nodal involvement and stages of tumor progression.Low risk HPV phenotypes could be one of the co-activators or/and co-carcinogens in complex, progressive, multifactorial and multistep esophageal carcinogenesis.

The study was undertaken to evaluate macrophage-derived chemokine (CCL22) levels in the peritoneal fluid (PF) and plasma of patients with ovarian cancer (n = 93) in relation to regulatory T cells (Tregs; n = 75). The peritoneal fluid CCL22 concentrations were significantly higher in epithelial ovarian cancer (EOC) patients than in patients with benign tumors-serous cystadenoma (n = 32). There was no difference in plasma levels of CCL22 in EOC patients compared with the non-cancer and healthy volunteers (n = 10). There were no significant differences in the plasma and PF CCL22 levels based on tumor grade. However, women with stage IV FIGO (International Federation of Gynecologists and Obstetricians) had significantly higher plasma CCL22 levels than patients with stages I and III. Women with stage I FIGO had significantly higher PF CCL22 levels than patients with stages II and III. Women with endometrioid cystadenocarcinoma had higher PF CCL22 levels than women with undifferentiated carcinoma. The percentage of tumor-infiltrating Tregs (11.06 %) was significantly higher compared to PF (3.05 %) and peripheral blood (PB) (2.01 %). Moreover, the percentage of Tregs was higher in the PF than in the PB of EOC patients. There were no significant differences in the PB, PF, and tumor-infiltrating Tregs percentage based on tumor stage, grade, or histology. Elevated levels of CCL22 found in the ascites could create a chemokine gradient aiding in Treg cells migration. Increased Tregs percentage in the local microenvironment of ovarian cancer might be an important mechanism of immunosuppression.