Publications by authors named "Iwona Wertel"

53 Publications

Lensoside Aβ as an Adjuvant to the Anti-Glioma Potential of Sorafenib.

Cancers (Basel) 2021 May 27;13(11). Epub 2021 May 27.

Department of Functional Anatomy and Cytobiology, Institute of Biological Sciences, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland.

Aim: The anti-glioma effect of lensoside Aβ alone and in combination with sorafenib (pro-survival Raf kinase inhibitor) was evaluated for the first time in terms of programmed cell death induction in anaplastic astrocytoma and glioblastoma multiforme cell lines as an experimental model. Apoptosis, autophagy, and necrosis were identified microscopically (fluorescence and scanning microscopes) and confirmed by flow cytometry (mitochondrial membrane potential MMP and cell death). The expression of apoptotic (caspase 3) and autophagic markers (beclin 1) as well as Raf kinase were estimated by immunoblotting. The FTIR method was used to determine the interaction of the studied drugs with lipid and protein groups within cells, while the modes of drug action within the cells were assessed with the FLIM technique.

Results: Lensoside Aβ itself does not exhibit anti-glioma activity but significantly enhances the anti-cancer potential of sorafenib, initiating mainly apoptosis of up to 90% of cells. It was correlated with an increased level of active caspase 3, a reduced MMP value, and a lower level of Raf kinase. The interaction with membrane structures led to morphological changes typical of programmed death.

Conclusions: Our results indicate that lensoside Aβ plays an important role as an adjuvant in chemotherapy with sorafenib and may be a potential candidate in anti-glioma combination therapy.
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http://dx.doi.org/10.3390/cancers13112637DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8198162PMC
May 2021

Involvement of PI3K Pathway in Glioma Cell Resistance to Temozolomide Treatment.

Int J Mol Sci 2021 May 13;22(10). Epub 2021 May 13.

Department of Functional Anatomy and Cytobiology, Institute of Biological Sciences, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland.

The aim of the study was to investigate the anticancer potential of LY294002 (PI3K inhibitor) and temozolomide using glioblastoma multiforme (T98G) and anaplastic astrocytoma (MOGGCCM) cells. Apoptosis, autophagy, necrosis, and granules in the cytoplasm were identified microscopically (fluorescence and electron microscopes). The mitochondrial membrane potential was studied by flow cytometry. The activity of caspases 3, 8, and 9 and Akt was evaluated fluorometrically, while the expression of Beclin 1, PI3K, Akt, mTOR, caspase 12, and Hsp27 was determined by immunoblotting. SiRNA was used to block Hsp27 and PI3K expression. Cell migration and localization of Hsp27 were tested with the wound healing assay and immunocytochemistry, respectively. LY294002 effectively diminished the migratory potential and increased programmed death of T98G and MOGGCCM. Autophagy was dominant in MOGGCCM, while apoptosis was dominant in T98G. LY294002 with temozolomide did not potentiate cell death but redirected autophagy toward apoptosis, which was correlated with ER stress. A similar effect was observed after blocking PI3K expression with siRNA. Transfection with Hsp27 siRNA significantly increased apoptosis related to ER stress. Our results indicate that inhibition of the PI3K/Akt/mTOR pathway sensitizes glioma cells to apoptosis upon temozolomide treatment, which was correlated with ER stress. Hsp27 increases the resistance of glioma cells to cell death upon temozolomide treatment.
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http://dx.doi.org/10.3390/ijms22105155DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8152763PMC
May 2021

The Role of Myeloid-Derived Suppressor Cells (MDSCs) in the Development and/or Progression of Endometriosis-State of the Art.

Cells 2021 Mar 18;10(3). Epub 2021 Mar 18.

Independent Laboratory of Cancer Diagnostics and Immunology, Department of Oncological Gynaecology and Gynaecology, Medical University of Lublin, Staszica 16, 20-081 Lublin, Poland.

Endometriosis (EMS) is a common gynecological disease characterized by the presence of endometrial tissue outside the uterus. Approximately 10% of women around the world suffer from this disease. Recent studies suggest that endometriosis has potential to transform into endometriosis-associated ovarian cancer (EAOC). Endometriosis is connected with chronic inflammation and changes in the phenotype, activity, and function of immune cells. The underlying mechanisms include quantitative and functional disturbances of neutrophils, monocytes/macrophages (MO/MA), natural killer cells (NK), and T cells. A few reports have shown that immunosuppressive cells such as regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) may promote the progression of endometriosis. MDSCs are a heterogeneous population of immature myeloid cells (dendritic cells, granulocytes, and MO/MA precursors), which play an important role in the development of immunological diseases such as chronic inflammation and cancer. The presence of MDSCs in pathological conditions correlates with immunosuppression, angiogenesis, or release of growth factors and cytokines, which promote progression of these diseases. In this paper, we review the impact of MDSCs on different populations of immune cells, focusing on their immunosuppressive role in the immune system, which may be related with the pathogenesis and/or progression of endometriosis and its transformation into ovarian cancer.
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http://dx.doi.org/10.3390/cells10030677DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003224PMC
March 2021

Gender Dysphoria Disrupting the Course of Treatment of a Recurrent Juvenile Granulosa Cell Tumor in an Adolescent Female: A Case Report.

Case Rep Oncol 2020 Sep-Dec;13(3):1330-1336. Epub 2020 Nov 10.

Independent Laboratory of Cancer Diagnostics and Immunology, Ist Chair and Department of Oncological Gynaecology and Gynaecology, Medical University of Lublin, Lublin, Poland.

We present the case of an adolescent female patient with gender dysphoria (GD) who was diagnosed with a recurrent ovarian neoplasm - juvenile granulosa cell tumor (JGCT). The 17-year-old female patient presented multiple endocrine pathologies and a recurrent JGCT. During the surgery qualification process, the patient admitted having identified herself as a male. The patient reported being uncomfortable with her body and with the expected roles of her assigned gender. Due to that, the patient requested a total hysterectomy with a bilateral salpingo-oophorectomy. As a minor, she required the permission of her parents, which was not granted. The patient underwent several specialist consultations, after which she agreed to the unilateral removal of tumor-changed pathologies and additional hormonal, psychological, and psychiatric diagnostics. To the best of our knowledge, this is the first detailed report of co-occurrence of GD spectrum disorders and JGCT in an adolescent female. This case contains many therapeutic and ethical problems regarding both physical and mental health. It should be noted that adolescents with GD spectrum rarely develop persistent transsexuality. Modulations from developmental psychology, psychotherapy, family dynamics, hormonal treatment, and the removal of JGCT in the presented case may have potential therapeutic implications for GD.
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http://dx.doi.org/10.1159/000510810DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7747049PMC
November 2020

Programmed Death-1 Receptor (PD-1) as a Potential Prognosis Biomarker for Ovarian Cancer Patients.

Cancer Manag Res 2020 7;12:9691-9709. Epub 2020 Oct 7.

Independent Laboratory of Cancer Diagnostics and Immunology, I Chair and Department of Oncological Gynaecology and Gynaecology, Medical University of Lublin, Lublin 20-081, Poland.

Aim: Ovarian cancer (OC) is one of the most lethal gynecological malignancies. Recent studies suggest a crucial role of the PD-1/PD-L1 pathway in OC pathogenesis. Therefore, our study aimed at evaluation of the clinical importance of PD-1 expression in ovarian cancer patients.

Patients And Methods: In this study, we investigated the role of PD-1 in OC patients (n=50) by analyzing its expression on CD4 and CD8 T cells in three OC environments: peripheral blood (PB), peritoneal fluid (PF), and tumor (TT) as well as soluble PD-1 (sPD-1) in plasma and PF in terms of their clinical and prognostic significance. T cells with PD-1 expression were analyzed using flow cytometry. The concentration of sPD-1 was determined with the use of ELISA. Our research demonstrated differences in PD-1 expression on CD4 and CD8 T cells in the OC environments.

Results: We found an elevated level of CD4PD-1 T cells in tumor and PF, compared to PB. Additionally, we found the highest percentage of CD8 PD-1 in tumor, compared to PB and PF. The levels of sPD-1 were higher (p<0.0001) in plasma than in PF. For the first time, we discovered that the higher level of CD4PD-1 T cells in the circulation and the higher sPD-1 level in plasma predict poor survival of OC patients.

Conclusion: We suggest that PD-1 could be a predictive biomarker for OC patients and successful immunotherapy.
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http://dx.doi.org/10.2147/CMAR.S263010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7548235PMC
October 2020

Prognostic and Clinical Value of Interleukin 6 and CD45CD14 Inflammatory Cells with PD-L1/PD-L2 Expression in Patients with Different Manifestation of Ovarian Cancer.

J Immunol Res 2020 30;2020:1715064. Epub 2020 Sep 30.

I Chair and Department of Oncological Gynaecology and Gynaecology, Medical University of Lublin, Staszica 16, Lublin 20-081, Poland.

Ovarian cancer (OC) is one of the deadliest gynecological cancers. Recent studies suggest a crucial role of inflammatory immune system cells in the progression and metastasis of OC. The understanding of inflammatory mechanisms is pivotal for the selection of a biomarker that allows the differentiation between malignant and benign tumors, monitoring the progression of the disease, and identification of patients that will respond to implemented treatment. Our study is aimed at evaluating the profile of IL-6 in the plasma and peritoneal fluid (PF) of patients with various clinical manifestations of OC ( = 78). We also examined the relationship between IL-6 and PD-L1/PD-L2 positive CD45CD14 inflammatory cell (MO/MA) levels in three OC environments (TME): peripheral blood (PB), PF, and tumor (TT) and their clinical and prognostic relevance in OC patients. The expression of PD-L1/PD-L2 molecules was analyzed by flow cytometry. The IL-6 levels were determined by ELISA. We found an elevated level of PD-L1/PD-L2 positive MO/MA in TT compared to PB ( < 0.0001). Significantly higher ( < 0.0001) levels of IL-6 were observed in PF of the OC patients than in the benign ovarian tumor group ( = 31). Additionally, we found higher IL-6 levels in PF than in the plasma of the OC patients. Interestingly, accumulation of IL-6 was observed in PF of patients with low-differentiated OC and correlated with worse prognosis. Moreover, we observed correlations between the level of IL-6 and CD45CD14 cells and between CD45CD14PD-L1 cells and the IL-6 level in PF. For the first time, we discovered that the higher percentage of CD45CD14PD-L2 cells in PF predicts better survival of OC patients. Our study suggests that CD45CD14PD-L2 cells and IL-6 may be predictive biomarkers for OC patients. Understanding how the composition of TME changes during OC development and progression is a prerequisite for projecting new therapeutic strategies. Overall, further validation research is warranted.
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http://dx.doi.org/10.1155/2020/1715064DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7545411PMC
September 2020

Cancer SLC43A2 alters T cell methionine metabolism and histone methylation.

Nature 2020 09 2;585(7824):277-282. Epub 2020 Sep 2.

Department of Surgery, University of Michigan School of Medicine, Ann Arbor, MI, USA.

Abnormal epigenetic patterns correlate with effector T cell malfunction in tumours, but the cause of this link is unknown. Here we show that tumour cells disrupt methionine metabolism in CD8 T cells, thereby lowering intracellular levels of methionine and the methyl donor S-adenosylmethionine (SAM) and resulting in loss of dimethylation at lysine 79 of histone H3 (H3K79me2). Loss of H3K79me2 led to low expression of STAT5 and impaired T cell immunity. Mechanistically, tumour cells avidly consumed methionine and outcompeted T cells for methionine by expressing high levels of the methionine transporter SLC43A2. Genetic and biochemical inhibition of tumour SLC43A2 restored H3K79me2 in T cells, thereby boosting spontaneous and checkpoint-induced tumour immunity. Moreover, methionine supplementation improved the expression of H3K79me2 and STAT5 in T cells, and this was accompanied by increased T cell immunity in tumour-bearing mice and patients with colon cancer. Clinically, tumour SLC43A2 correlated negatively with T cell histone methylation and functional gene signatures. Our results identify a mechanistic connection between methionine metabolism, histone patterns, and T cell immunity in the tumour microenvironment. Thus, cancer methionine consumption is an immune evasion mechanism, and targeting cancer methionine signalling may provide an immunotherapeutic approach.
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http://dx.doi.org/10.1038/s41586-020-2682-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7486248PMC
September 2020

Extracts Affect the Viability and Proliferation of Non-Cancerous and Cancerous Colon Human Epithelial Cells.

Molecules 2020 Jul 6;25(13). Epub 2020 Jul 6.

Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Białystok, ul. Mickiewicza 2a, 15-230 Białystok, Poland.

The aim of this study was to determine the anti-tumor activity of extracts isolated from L. on human colon cancer cells of the HT-29 line and on non-cancer colon epithelial cells of the CCD 841 CoTr line. The research methods we used to determine the cytotoxic and proliferative properties were 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red (NR) assays, the ability to produce nitric oxide, the Griess method, and the biochemical properties like 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) methods indicating reduction activity of tested samples. Finally, the effects of the extracts on the morphology and cell counts were assessed by May-Grünwald-Giemsa staining. After a comprehensive analysis of all the experiments, the extracts were found to demonstrate cytotoxic properties, they stimulated the division of non-cancer cells, and they were able to scavenge free radicals. In the NR method, the cell viability dropped to approximately 80% compared to the control. In the MTT assay, tumor cell proliferation decreased to 9.5% compared to the control. Therefore, we concluded that this plant has medical potential.
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http://dx.doi.org/10.3390/molecules25133080DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7411782PMC
July 2020

Coumarins modulate the anti-glioma properties of temozolomide.

Eur J Pharmacol 2020 Aug 22;881:173207. Epub 2020 May 22.

Department of Functional Anatomy and Cytobiology, Maria Curie-Sklodowska University, Akademicka 19, 20-033, Lublin, Poland. Electronic address:

In the recent years, coumarin bioactive compounds have been identified to posess anticancer properties. Therefore, the aim of the present study was to investigate for the first time the efficacy of osthole, umbelliferone, esculin, and 4-hydroxycoumarin, alone and in combination with Temozolomide, in the elimination of deadly brain tumors, anaplastic astrocytoma (AA) and glioblastoma multiforme (GBM) cells via programmed death. Our results indicated that osthole, umbelliferone, esculin, and 4-hydroxycoumarin initiated mainly apoptosis in the T98G and MOGGCCM cells. Osthole was the most effective. It also initiated autophagy in a small percentage of the cell population. The co-incubation with Temozolomide did not increase the pro-apoptotic potential of natural compounds but decreased the level of autophagy in the T98G cells. Apoptosis was associated with reduced mitochondrial membrane potential, activation of caspase 3, inhibition of Bcl-2 expression and the presence of a Bcl-2/Beclin 1. Blocking of Bcl-2 expression resulted in promotion of apoptosis, but not autophagy, in the MOGGCCM and T98G lines. It also sensitized astrocytoma cells, but not GBM, to the combined osthole and TMZ treatment, which was correlated with a reduced level of Beclin 1 and increased expression of caspase 3. Osthole and TMZ, alone and in combination, inhibited the migratory phenotype of the GBM and AA cells. In summary, our results indicated that osthole effectively eliminated glioma cells via apoptosis, what was correlated with Bcl-2/Beclin 1 complex formation. Considering the anti-migratory effect, osthole and Temozolomide display antiglioma potential but it needs further extensive studies.
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http://dx.doi.org/10.1016/j.ejphar.2020.173207DOI Listing
August 2020

Th17 Cells and IL-17 As Novel Immune Targets in Ovarian Cancer Therapy.

J Oncol 2020 21;2020:8797683. Epub 2020 Feb 21.

Independent Laboratory of Cancer Diagnostics and Immunology, The First Department of Gynecologic Oncology and Gynaecology, Medical University of Lublin, Staszica 16, Lublin 20-081, Poland.

Ovarian cancer (OC) is usually diagnosed at an advanced stage and is related with poor prognosis. Despite numerous studies, the pathogenesis of OC is still unknown. Recent studies indicate the role of the immune system in the development and spread of OC. The identification of factors and mechanisms involved in that process and their modulation is crucial for creating effective antitumor therapy. We investigated the potential role of Th17 cells in OC patients ( = 71) by analyzing the frequencies of Th17 cells in three different environments, i.e., peripheral blood (PB), peritoneal fluid (PF), and tissue (Th17 infiltrating cells), and the concentration of IL-17A in plasma and PF of patients in terms of their clinical and prognostic significance. Th17 cells were analyzed by flow cytometry as a percentage of CD4 lymphocytes that expressed intracellular expression of IL-17A. The level of IL-17A in plasma and PF were determined by ELISA. Our results showed accumulation of Th17 cells among tumor-infiltrating CD4 lymphocytes ( < 0.001 in relation to PB). Moreover, the percentage of Th17 cells in both PB and PF of OC patients was significantly lower than that in benign tumors group ( = 35). There were no significant differences in the percentage of Th17 cells in PB, PF, and tissue in relation to clinicopathological characteristics of OC patients and survival. The lower percentage of Th17 cells in the PB and PF of OC patients may promote evasion of host immune response by cancer cells. The concentration of IL-17A in plasma of OC patients was higher ( < 0.0001) than that in both benign tumors and control group ( = 10). The PF IL-17A level in OC patients was higher ( < 0.0001) than that in women with benign ovarian tumors, indicating its synthesis in OC microenvironment. Higher IL-17A level in PF is correlated with longer (median: 36.5 vs. 27 months) survival of OC patients.
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http://dx.doi.org/10.1155/2020/8797683DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7054820PMC
February 2020

[Selected mechanisms inducing resistance to immunotherapy in patients with ovarian cancer].

Wiad Lek 2019 ;72(7):1397-1402

Pracownia Immunologii Nowotworów, i Katedra i Klinika Ginekologii Onkologicznej i Ginekologii, Uniwersytet Medyczny w Lublinie, Lublin, Polska.

Recently, the intensive development of immunotherapies in the treatment of malignant tumors has been observed. The investigated treatment approaches including specific monoclonal antibodies, adoptive therapy and also anticancer vaccinations. The implementation of immunotherapy seems to be promising in treatment of the most malignant and fatal tumors including ovarian cancer. However, current findings have shown only a nonsignificant improvement of patients' survival. The possible cause of failure may be immunotherapy barriers that are a result of low immunogenicity level of ovarian cancer cells, mutation variability, and also the presence of a specific, immunosuppressive tumor microenvironment, which stimulates the cancer progression. The review presents the selected mechanisms of tumor resistance to immunological therapy. In order to project effective treatment approaches, it is necessary to understand both, mechanisms leading to the correct response for the treatment and causing therapeutic failures, resulting from resistance to therapy.
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August 2019

Clinical Relevance and Immunosuppressive Pattern of Circulating and Infiltrating Subsets of Myeloid-Derived Suppressor Cells (MDSCs) in Epithelial Ovarian Cancer.

Front Immunol 2019 3;10:691. Epub 2019 Apr 3.

The First Department of Gynecologic Oncology and Gynecology, Medical University of Lublin, Lublin, Poland.

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.
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http://dx.doi.org/10.3389/fimmu.2019.00691DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6456713PMC
July 2020

[Characterization of chosen pro-angiogenic factors and anti-angiogenic therapies in ovarian cancer].

Wiad Lek 2018;71(8):1603-1607

I Katedra i Klinika Ginekologii Onkologicznej I Ginekologii Uniwersytetu Medycznego W Lublinie, Lublin, Polska.

Ovarian cancer is a neoplasm characterized by notably malignancy. The poor results of treatment result not only from the lack of screening tests making diagnosis possible at an early stage, but also because of the insufficiently effective treatment methods. High hopes are placed in targeted therapies, using agents such as angiogenesis inhibitors, immune checkpoint inhibitors or anticancer vaccines. The aim of the work is to present the latest results of research on antiangiogenic drugs. Databases such as PubMed, Google Scholar and ClinicalTrials.gov were used. Antiangiogenic drugs are substances of various structure, the common feature of which is the influence on signaling pathways associated with such factors as VEGF, PDGF or Ang1 / 2. Bevacizumab is an antibody directed against VEGF-A. It is the first anti-angiogenic drug with proven efficacy, expressed in the extension of overall survival. This was demonstrated both in the group of patients with newly diagnosed advanced disease and in the situation of relapse. Other anti-angiogenic agents, such as trebananib, nintedanib or pazopanib, currently have not been proven to possess comparably high efficacy in the treatment of ovarian cancer. There are no known causes of disease progression despite maintenance therapy. The potential for combining bevacizumab with other targeted drugs such as PD-L1 inhibitors is currently being investigated.
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June 2019

The Effect of Fucoidan, a Potential New, Natural, Anti-Neoplastic Agent on Uterine Sarcomas and Carcinosarcoma Cell Lines: ENITEC Collaborative Study.

Arch Immunol Ther Exp (Warsz) 2019 Apr 18;67(2):125-131. Epub 2019 Jan 18.

1st Chair and Department of Gynecological Oncology and Gynecology, Medical University of Lublin, Staszica 16, 20-081, Lublin, Poland.

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.
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http://dx.doi.org/10.1007/s00005-019-00534-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6420609PMC
April 2019

Disrupted iron metabolism in peritoneal fluid may induce oxidative stress in the peritoneal cavity of women with endometriosis.

Ann Agric Environ Med 2018 Dec 11;25(4):587-592. Epub 2017 Jul 11.

Department of Oncological Gynecology and Gynecology, Medical University of Lublin.

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.
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http://dx.doi.org/10.26444/aaem/75802DOI Listing
December 2018

Assessment of the clinicopathological relevance of mesothelin level in plasma, peritoneal fluid, and tumor tissue of epithelial ovarian cancer patients.

Tumour Biol 2018 Oct;40(10):1010428318804937

1 Tumor Immunology Laboratory, The First Department of Gynecologic Oncology and Gynecology, Medical University of Lublin, Lublin, Poland.

Ovarian cancer remains the most lethal gynecologic malignancy. This is due to lack of effective screening, diagnosis predominance in late stage of disease, a high recurrence rate after primary therapy, and poor treatment response in platinum-resistant tumor. Thus, unique biomarkers, predictive of individual disease course, and prognosis are urgently needed. The aim of our study was to assess the clinicopathological significance of plasma, peritoneal fluid, and tumor tissue levels of mesothelin in epithelial ovarian cancer patients. Plasma and peritoneal fluid levels of mesothelin were measured by enzyme-linked immunosorbent assay. Tissue expression of MSLN was evaluated using quantitative real-time polymerase chain reaction. Preoperative plasma mesothelin levels were significantly higher in epithelial ovarian cancer patients in comparison to the patients with benign tumor and controls. There have been noticed significant differences in the plasma mesothelin levels based on International Federation of Gynecology and Obstetrics stage, grade, and histology type. No significant changes were observed between Kurman and Shih type I versus type II epithelial ovarian cancer. Interestingly, peritoneal fluid mesothelin levels revealed significant differences based on both grade and Kurman and Shih-type epithelial ovarian cancer. There were no relevant changes in the mesothelin level in peritoneal fluid between different stages and histology types compared to benign tumor. MSLN expression level in tumor tissue was significantly higher based on stage, grade, and Kurman and Shih-type epithelial ovarian cancer than in the benign masses. In addition, data showed significant higher MSLN expression in endometrioid tumors compared to benign masses and serous tumors. Plasma, peritoneal fluid, and tumor tissue levels of mesothelin positively correlated with level of CA125. Low mesothelin concentrations in plasma were also associated with prolonged patient survival. More importantly, we revealed that plasma mesothelin level was correlated with both peritoneal fluid mesothelin level and tumor MSLN expression. This study highlights that plasma mesothelin level may be a useful noninvasive biomarker surrogate for local tumor mesothelin status in monitoring of epithelial ovarian cancer patients.
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http://dx.doi.org/10.1177/1010428318804937DOI Listing
October 2018

[The meaning of PD-1/PD-L1 pathway in ovarian cancer pathogenesis].

Wiad Lek 2018;71(5):1089-1094

I Katedra I Klinika Ginekologii Onkologicznej I Ginekologii Uniwersytetu Medycznego W Lublinie, Lublin, Polska.

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.
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March 2019

Blood-based analyses of cancer: Circulating myeloid-derived suppressor cells - is a new era coming?

Crit Rev Clin Lab Sci 2018 09 21;55(6):376-407. Epub 2018 Jun 21.

a 1st Chair and Department of Oncological Gynaecology and Gynaecology, Tumor Immunology Laboratory , Medical University of Lublin , Lublin , Poland.

Progress in cancer treatment made by the beginning of the 21st century has shifted the paradigm from one-size-fits-all to tailor-made treatment. The popular vision, to study solid tumors through the relatively noninvasive sampling of blood, is one of the most thrilling and rapidly advancing fields in global cancer diagnostics. From this perspective, immune-cell analysis in cancer could play a pivotal role in oncology practice. This approach is driven both by rapid technological developments, including the analysis of circulating myeloid-derived suppressor cells (cMDSCs), and by the increasing application of (immune) therapies, the success or failure of which may depend on effective and timely measurements of relevant biomarkers. Although the implementation of these powerful noninvasive diagnostic capabilities in guiding precision cancer treatment is poised to change the ways in which we select and monitor cancer therapy, challenges remain. Here, we discuss the challenges associated with the analysis and clinical aspects of cMDSCs and assess whether the problems in implementing tumor-evolution monitoring as a global tool in personalized oncology can be overcome.
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http://dx.doi.org/10.1080/10408363.2018.1477729DOI Listing
September 2018

Temozolomide and sorafenib as programmed cell death inducers of human glioma cells.

Pharmacol Rep 2017 Aug 14;69(4):779-787. Epub 2017 Mar 14.

Department of Medical Biology, Institute of Agricultural Medicine, Lublin, Poland; Department of Immunology and Virology, Maria Curie-Sklodowska University, Lublin, Poland. Electronic address:

Background: Gliomas are aggressive brain tumors with very high resistance to chemotherapy. Therefore, the aim of the present study was to investigate the effectiveness of sorafenib and Temozolomide in elimination of human glioma cells through apoptosis and autophagy.

Methods: MOGGCCM (anaplastic astrocytoma) and T98G (glioblastoma multiforme) cell lines incubated with sorafenib and/or Temozolomide were used in the experiments. Cell morphology (ER stress, apoptosis, autophagy, and necrosis) was analyzed microscopically while apoptosis and mitochondrial membrane potential were assessed with flow cytometry. Beclin1, LC3, p62, Hsp27, and Hsp72 levels were analyzed by immunoblotting. The activity of caspase 3, 8, and 9 was evaluated fluorometrically. Expression of Hsps was blocked by transfection with specific siRNA.

Results: In MOGGCCM cells, Temozolomide most frequently induced autophagy, which was accompanied by decreased p62 and increased beclin1 and LC3II levels. Sorafenib initiated mainly apoptosis. Additional incubation with Temozolomide, synergistically potentiated the pro-apoptotic properties of sorafenib, but it was mediated in a caspase-independent way. In T98G cells, the effect of the analyzed drugs on programmed cell death induction was different from that in MOGGCCM cells. Sorafenib induced autophagy, while Temozolomide initiated mainly apoptosis. After simultaneous drug application, apoptosis dominated, suggesting synergistic action of both drugs. Inhibition of Hsp27 and Hsp72 expression increased the sensitivity of both cell lines to ER stress and, to a lesser extent, to induction of apoptosis, but not autophagy.

Conclusions: Sorafenib and Temozolomide applied in combination are potent apoptosis inducers in T98G and MOGGCCM cells. ER stress precedes the elimination. Blocking of Hsp expression has a greater impact on ER stress rather than apoptosis induction.
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http://dx.doi.org/10.1016/j.pharep.2017.03.008DOI Listing
August 2017

Does the patients age at cancer diagnosis affect microvessels density in uterine sarcoma tissues?

Ginekol Pol 2017 ;88(3):138-140

1st Chair and Department of Gynecological Oncology and Gynecology, Medical University of Lublin, Poland.

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-de-pendent 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 (MVDCD34).

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.
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http://dx.doi.org/10.5603/GP.a2017.0026DOI Listing
July 2018

[Why ovarian cancer cells escape from immune surveillance?]

Wiad Lek 2017;70(1):74-80

I Katedra I Klinika Ginekologii Onkologicznej I Ginekologii Uniwersytetu Medycznego, Lublin, Polska.

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.
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July 2017

Management of uterine leiomyosarcoma.

Wiad Lek 2016;69(6):799-803

Katedra i Klinika Ginekologii Onkologicznej i Ginekologii, Uniwersytet Medyczny, Lublin, Polska.

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.
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July 2017

Tumor-Associated Macrophages and Myeloid-Derived Suppressor Cells as Immunosuppressive Mechanism in Ovarian Cancer Patients: Progress and Challenges.

Int Rev Immunol 2016 09 19;35(5):372-385. Epub 2016 Sep 19.

a Department of Oncological Gynaecology and Gynaecology , Medical University , Lublin , Poland.

Cancers are complex masses of malignant cells and nonmalignant cells that create the tumor microenvironment (TME). Non-transformed cells of the TME such as tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) have been observed in the TME of ovarian cancer (OC) patients. Although these subsets may contribute to each step of carcinogenesis and are commonly associated with poor prognosis, still little is known about creation of the protumor microenvironment in OC. In this review, we focused on the nature and prognostic significance of TAMs and MDSCs in OC patients. Moreover, we discuss the main problems and challenges that must be overcome by researchers and clinicians to enrich our knowledge about the immunosuppressive microenvironment of cancers.
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http://dx.doi.org/10.1080/08830185.2016.1206097DOI Listing
September 2016

[Assessment of regulatory T cells, cytotoxic lymphocytes and dendritic cells in ovarian cancer patients before and after menopause].

Ginekol Pol 2016 ;87(1):11-8

I Katedra i Klinika Ginekologii Onkologicznej i Ginekologii., I Katedra i Klinika Ginekologii Onkologicznej i Ginekologii, Uniwersytet Medyczny w Lublinie.

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http://dx.doi.org/10.17772/gp/60976DOI Listing
July 2018

[The prognostic value of selected immunological factors in ovarian cancer patients].

Wiad Lek 2015 ;68(4 Pt 2):690-4

I Katedra i Klinika Ginekologii Onkologicznej i Ginekologii, Uniwersytet Medyczny, Lublin.

Ovarian cancer is the most aggressive gynecological cancer and is often diagnosed in advanced stage. Constantly we are looking for new prognostic factors which would enable early diagnosis, increase the effectiveness of therapeutic intervention. There is to little data about immunological predictors in ovarian cancer. The tumor's microenvironment is designated by regulatory T cells, cytotoxic T cells, dendritic cells (DCs), tumor - associated macrophages (TAMs), monocytes, plasma cells and cytokines, such as IL-6, IL-8, IL-10, IL-17 and TGF-beta. Some of them are responsible for the inhibition and others induce tumor growth. Ovarian cancer patients with high ratio of CD8 + TILs to Treg present longer overall survival time (OS). The presence of T helper cells in ascites is associated with longer OS. Furthermore, patients with a lower rate plasmocytoid DCs infiltrating tumor tissue demonstrate longer progression-free survival time (PFS). Women with increased M1/M2 ratio present higher five-year survival rate. The presence of immunologically competent cells and secreted cytokines give motivation to evaluate their prognostic value. Perhaps this strategy will contribute to longer progression-free survival time and overall survival time in those patients.
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June 2016

Quercetin and sorafenib as a novel and effective couple in programmed cell death induction in human gliomas.

Neurotox Res 2014 Jul 24;26(1):64-77. Epub 2013 Dec 24.

Department of Comparative Anatomy and Anthropology, Maria Curie-Skłodowska University, Akademicka 19, 20-033, Lublin, Poland,

The aim of the present study was to investigate the effect of sorafenib and quercetin on the induction of apoptosis and autophagy in human anaplastic astrocytoma (MOGGCCM) and glioblastoma multiforme (T98G) cell lines. In MOGGCCM cells, sorafenib initiated mainly apoptosis, mediated by the mitochondrial pathway with mitochondrial membrane permeabilization, cytochrome c release to the cytoplasm, and activation of caspase 9 and 3. Additional incubation with quercetin potentiated the pro-apoptotic properties of sorafenib. In T98G cells, autophagy was observed most frequently after the sorafenib treatment. It was accompanied by increased beclin 1 and LC3II expression. Administration of quercetin after the sorafenib treatment resulted in an increased number of autophagic cells. After simultaneous drug application, the level of autophagy was lower in favour of apoptosis. Inhibition of heat shock proteins expression by specific small interfering RNA significantly increased the sensitivity of both the cell lines to induction of apoptosis, but not autophagy. We demonstrated for the first time that sorafenib and quercetin are very effective programmed cell death inducers in T98G and MOGGCCM cells, especially in cells with blocked expression of heat shock proteins.
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http://dx.doi.org/10.1007/s12640-013-9452-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4035551PMC
July 2014

Silencing of Hsp27 and Hsp72 in glioma cells as a tool for programmed cell death induction upon temozolomide and quercetin treatment.

Toxicol Appl Pharmacol 2013 Dec 12;273(3):580-9. Epub 2013 Oct 12.

Department of Comparative Anatomy and Anthropology, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland. Electronic address:

The aim of the present study was to investigate whether silencing of Hsp27 or Hsp72 expression in glioblastoma multiforme T98G and anaplastic astrocytoma MOGGCCM cells increases their sensitivity to programmed cell death induction upon temozolomide and/or quercetin treatment. Transfection with specific siRNA was performed for the Hsp gene silencing. As revealed by microscopic observation and flow cytometry, the inhibition of Hsp expression was correlated with severe apoptosis induction upon the drug treatment studied. No signs of autophagy were detected. This was correlated with a decreased mitochondrial membrane potential, increased level of cytochrome c in the cytoplasm, and activation of caspase 3 and caspase 9. All these results suggest that the apoptotic signal was mediated by an internal pathway. Additionally, in a large percentage of cells treated with temozolomide, with or without quercetin, granules within the ER system were found, which was accompanied by an increased level of caspase 12 expression. This might be correlated with ER stress. Quercetin and temozolomide also changed the shape of nuclei from circular to "croissant like" in both transfected cell lines. Our results indicate that blocking of Hsp27 and Hsp72 expression makes T98G cells and MOGGCCM cells extremely vulnerable to apoptosis induction upon temozolomide and quercetin treatment and that programmed cell death is initiated by an internal signal.
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http://dx.doi.org/10.1016/j.taap.2013.10.003DOI Listing
December 2013

Low-density lipoproteins oxidation and endometriosis.

Mediators Inflamm 2013 19;2013:624540. Epub 2013 Jun 19.

1st Department of Oncological Gynecology and Gynecology, Medical University of Lublin, Staszica 16, 20-081 Lublin, Poland.

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 (n = 78) or dermoid (n = 41) 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.
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http://dx.doi.org/10.1155/2013/624540DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3703791PMC
February 2014

Apoptosis induction in human glioblastoma multiforme T98G cells upon temozolomide and quercetin treatment.

Tumour Biol 2013 Aug 12;34(4):2367-78. Epub 2013 Apr 12.

Department of Comparative Anatomy and Anthropology, Maria Curie-Skłodowska University, Akademicka 19, 20-033, Lublin, Poland.

Glioblastoma multiforme is the most aggressive primary brain tumour. At the cellular and molecular levels, several mechanisms responsible for apoptosis or autophagy induction are blocked. Identification of molecular targets stimulating cells to initiate programmed cell death should be performed for therapeutic purposes. A promising solution is the combination of temozolomide and quercetin. The aim of our study was to evaluate the effect of both drugs, applied alone and in combinations, on apoptosis and autophagy induction in human glioblastoma multiforme T98G cells. Our results clearly indicate that quercetin and temozolomide induce apoptosis very significantly, having no effect on autophagy induction. At the molecular level, it was correlated with caspase 3 and 9 activation, cytochrome c release from the mitochondrium and a decrease in the mitochondrial membrane potential. Both drugs are also potent Hsp27 and Hsp72 inhibitors. This suggests that the apoptotic signal goes through an internal pathway. Increased expression of caspase 12 and the presence of several granules in the cytoplasm after temozolomide treatment with or without quercetin preceding appearance of apoptosis may suggest that apoptosis is initiated by ER stress. Additionally, it was accompanied by changes in the nuclear morphology from circular to 'croissant like'.
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http://dx.doi.org/10.1007/s13277-013-0785-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3713258PMC
August 2013

[The role of iron metabolism and oxidative stress in the pathogenesis of endometriosis].

Ginekol Pol 2013 Jan;84(1):62-4

Katedra i Klinika Ginekologii Onkologicznej i Ginekologii UM w Lublinie, Lublin, Polska.

Despite many years of extensive investigations and increasing number of studies, the pathogenesis of endometriosis remains unclear Accumulated data suggests that disrupted iron metabolism may induce oxidative stress in the peritoneal cavity of endometriosis patients.
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http://dx.doi.org/10.17772/gp/1542DOI Listing
January 2013