Experimental data show that superoxide dismutase 2 (SOD2) is involved in ochratoxin (OTA)-induced nephrotoxicity, whereas clinical data indicate the role of
rs4880 or glutathione peroxidase 1 (
...rs1050450 polymorphisms in end-stage renal disease and urothelial carcinoma risk, known to be the major complications of Balkan endemic nephropathy (BEN). Therefore, we hypothesized that
and
gene polymorphisms would influence the risk of BEN and its associated tumors.
The study was conducted in 207 BEN patients and 86 controls from endemic areas.
Individuals with both copies of variant
allele, known for lower mitochondrial antioxidant protection, are at a significantly higher BEN risk (OR = 2.6,
= 0.021). No association was observed between
gene polymorphism and BEN risk. Combining
and
genotypes did not alter the risk of BEN development. Regarding the risk of urothelial tumors in BEN patients, none of the polymorphisms studied was significantly associated with the risk of these tumors.
Polymorphism in
rs4880 gene affects the risk of BEN development. Hence,
genotyping could, together with a panel of other enzymes, be used as a biomarker of susceptibility in BEN areas.
Chronic lymphocytic leukemia (CLL) develops due to an imbalance between apoptosis and proliferation of B lymphocytes. Chrysin induced apoptosis in leukemia cell lines such as U937, MO7e, THP-1 and ...HL-60, but there has not yet been data demonstrating the apoptotic effect of chrysin on CLL cells. Therefore, in our investigation we examined the cytotoxicity of chrysin against two leukemia cell lines, MOLT-4 and JVM-13, peripheral blood lymphocytes isolated from B-CLL patients and peripheral blood mononuclear cells (PBMC) from healthy individuals in vitro. The effect of chrysin on viability of MOLT-4 and JVM-13 cell lines, B-CLL cells derived from 28 patients and PBMC from 16 healthy subjects was determined by MTT assay. The type of cell death induced by chrysin was verified by Annexin V/7AAD assay and acridine orange and ethidium bromide (AO/EB) staining assay. Intracellular localisation and endogenic expression of apoptotic proteins including Bax, Bcl-2, cytochrome c and caspase-3 were determined by flow cytometry and fluorescent microscopy. Our results demonstrated that exposure of MOLT-4, JVM-13 cell lines and B-CLL cells to the concentration of chrysin of 10μM and higher selectively decreased viability of cells in this cell population, but not in the PBMC derived from healthy subjects; LC50 values of chrysin for B-CLL cells were 51μM for 24 hours and 32μM for 48 hours of incubation, respectively. Our findings demonstrated that chrysin induces the activation of proapoptotic Bax and decreases the expression of antiapoptotic Bcl-2 protein, releases cytochrome c from mitochondria into cytosol and cleavages/activates caspase-3, subsequently leading to the activation of apoptosis of B-CLL cells. Together, these findings suggest that chrysin selectively induces apoptosis of peripheral blood lymphocytes isolated from human chronic lymphocytic leukemia patients via mitochondrial pathway in vitro and that it might have a promising role as a potential future antileukemic remedy.
Endometrial hyperplasia is a condition that occurs as a result of hormonal imbalance between estrogen and progesterone. Morphological disturbance of endometrial cells occurs consequently leading ...towards endometrial cancer. In therapy of endometrial hyperplasia SERMs are used to supress effects of locally high estrogen level in uterus. There is strong evidence suggesting that estrogen could be involved in cell death - apoptosis. There are no experimental data demstrating the direct apoptotic effect of both raloxifene and estrogen on the ThESC cell line. The aim of our study wa sto investigate both cytotoxic and apototic mechanism of raloxifene and estrogen - induced death in the ThESC cell line.
In order to determine their cytotoxic and apoptotic effects, various doses of raloxifene and estrogen were applied to the ThESC cell line for 24 h. After the treatment MTT assay, FACS analysis and immunofluoroscence method were conducted.
The results of this study for the first time demonstrated the cytotoxic and apoptotic effects of raloxifene and estrogen on human endometrial stromal cell line suggesting the involvement of the inner, mitochondrial apoptotic pathway.
Our results demonstrated apoptotic effects of investigated drugs in the ThESC cell line through increasing the Bax/Bcl-2 ratio and activation of caspase 3.
Background/Aim. The main cause for development of endometrial hyperplasia is unopposed effect of estrogen on endometrial cells. The aim of our study was to investigate and compare cytotoxic and ...apoptotic effects of mifepristone on human endometrial stromal cell line for the first time. Both percentage of cytotoxic and apoptotic cells were determined after 24 h treatment with different doses of mifepristone. Methods. The percentage of cytotoxic cell was evaluated by viability assay while the percentage of apoptotic cells was determined using flow cytometry. Determination of apoptotic effects was confirmed using immunofluorescence method determining expression and localization of active Bax and Bcl-2 proteins. Results. Our results indicated that mifepristone induced cytotoxic and apoptotic effect on human endometrial stromal cell line (ThESC) through changes in expression level of Bcl-2 and active Bax proteins. Conclusion. Cytotoxic and pro-apoptotic effects of mifepristone on human endometrial stromal cell line in vitro was investigated in this study for the first time. It is crucial to point out that mifepristone expressed both cytotoxic and pro-apoptotic effect on ThESC cell line. Our results may contribute to determination of localization and expression level of the crucial proteins involved in apoptosis in ThESC cell line after the treatment with the lowest cytotoxic doses of mifepristone.
Background/Aim. Chronic lymhocytic leukemia (CLL) is considered more as a disease of cells accumulation due to the defect in apoptosis rather than deregulated cell?s proliferation. The activation of ...apoptosis is one of the main molecular mechanisms responsible for anti-cancer activities of most of the currently studied potential anti-cancer agents, including natural compounds. Teucrium polium (TP) extracts exhibited strong cytotoxic effects in murine leukemia cell line, RAW 264.7 and human melanoma cell line, C32, but their cytotoxic effects against human leukemia cells were unknown. Methods. The viability of human leukemia cell lines (MOLT 4 and JVM 13), lymphocytes isolated from 28 patients with CLL (CLL cells), and peripheral blood mononuclear cells (PBMCs) isolated from 16 healthy subjects treated with TP leaves methanolic extract, was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis of TP treated CLL cells was measured by flow cytometry applying Annexin V/7AAD staining. The expressions of active proapoptotic protein Bax, antiapoptotic protein Bcl-2, cytochrome c and the percentage of cells containing cleaved caspase-3 in treated CLL cells was determined by flow cytometry and immunocytochemistry. Results. The TP meth-anolic extract decreased the viability of all tested human leukemia cells but it had no effect on the viability of PBMCs isolated from healthy subjects. The cytotoxic effect of TP was caused by its induction of CLL cells? apoptosis. TP disarranged the ratio of the expressions of proapoptotic Bax and antiapoptotic Bcl-2 protein in favor of Bax, consequently inducing apoptosis by cytochrome c mitochondrial release and activation of caspase-3 in treated CLL cells. Conclusion. The TP leaves methanolic induced selective apoptosis in CLL cells and it affected the expressions of key proteins involved in the regulation of programmed cell death.
Background & Aims We used Concanavalin A-induced liver injury to study the role of Interleukin 33 and its receptor ST2 in the induction of inflammatory pathology and hepatocellular damage. Methods We ...tested susceptibility to Concanavalin A induced hepatitis in ST2 deficient and wild type BALB/c mice and analyzed the effects of single injection of Interleukin 33 as evaluated by liver enzyme test, quantitative histology, mononuclear cell infiltration, cytokine production, intracellular staining of immune cells, and markers of apoptosis in the liver. Results ST2 deficient mice developed significantly more severe hepatitis and had significantly higher number of mononuclear cells in the liver, CD4+ and CD8+ T cells, NKp46+ and CD3+NKp46+ cells, and F4/80+ macrophages. The level of pro-inflammatory cytokines in the sera and number of TNF alpha, IFN gamma, and IL-17 producing cells was higher in ST2 deficient mice. In contrast, number of CD4+Foxp3+ cells was statistically higher in wild type mice. Additionally, treatment of wild type mice with single (1 μg) injection of Interleukin 33 led to attenuation of the liver injury and milder infiltration of mononuclear cells, increase in total number of liver CD4+Foxp3+ cells and IL-4 producing CD4+ T cells. Interleukin 33 also suppressed the activation of caspase 3, prevented the expression of BAX, and enhanced the expression of antiapoptotic Bcl-2 in the liver. Conclusions We concluded that Interleukin 33/ST2 axis downregulated Concanavalin A-induced liver injury and should be evaluated as potential target in fulminant hepatitis in humans.
•Cytokine levels in patients group were heterogeneous, spread in wide range of values.•Grouping data according to disease-relevant factors showed high intragroup diversity.•There was no correlation ...between cytokine levels and each disease-relevant factor.•Inconsistency of literature data is a result of individual differences of patients.
The data addressing cytokine profile in chronically infected HCV patients are conflicting, ranging from Th1 or Th2 cytokine prevalence to the expression of both types of cytokines. Therefore, the aim of this study was to evaluate cytokine profile in these patients. Cytokine sera levels in HCV patients and healthy controls were evaluated using 13plex FlowCytomix Multiplex. Median values of both proinflammatory and anti-inflammatory cytokines were lower in HCV patients then in controls. In addition, the number of subjects producing detectable quantities of cytokines was significantly lower in the group of HCV patients. Yet, cytokine levels in those patients were remarkably heterogeneous ranging from low to extremely high, much higher than the maximal values in control group. Similarly, grouping data according to HCV genotype, HCV RNA load, ALT/AST ratio and the stage of fibrosis showed marked standard deviations, reflecting high intragroup diversity. No correlation was found between each disease-related factor and cytokine levels. Patients investigated in our and similar studies were disparate pursuant to characteristics of the hosts, pathogen and course of the disease. Therefore, the inconsistency of the literature data regarding cytokine pattern in chronic HCV patients may be a consequence of the disregarded/overlooked heterogeneity of these patients.
Endometrial hyperplasia is a condition that may lead to the development of endometrial carcinoma. Initially, changes of the endometrium are caused by the estrogen's hyperstimulation that may lead to ...the development of an irregular bleeding and the infertility problems. Therapy of endometrial hyperplasia is limited to medical and surgical approaches. During the past decade, the new types of drugs were developed for the treatment of the endometrial hyperplasia. Here, for the first time, we investigated the cytotoxic effects of the various combinations of estrogen, raloxifene, and methotrexate in human ThESC cell line as a possible potential treatment of the endometrial hyperplasia. Our aim was to investigate and to determine the most efficient combination of investigated drugs in ThESC cells during 24‐hr period using MTT assay, FACS analysis, and immunofluorescence staining. Our results demonstrated that the combination of raloxifene with methotrexate efficiently induced both the cytotoxicity and apoptosis in ThESC cells when compared to their single effect, as well as to the effect of combined treatment of raloxifene with estrogen. The application of the low doses of methotrexate combined with raloxifene offers all advantages of a potential beneficial antitumor match in cancer chemoprevention and therapy.
Our results demonstrated that the combination of raloxifene with methotrexate more efficiently induced both the cytotoxicity and apoptosis in ThESC cells when compared to their single effect, as well as to the effect of combined treatment of raloxifene with estrogen. The application of the low doses of methotrexate combined with raloxifene offers all advantages of a potential beneficial antitumor match in cancer chemoprevention and therapy.
The treatment options of endometrial hyperplasia consist of surgical, interventional and medical therapies including apoptosis-inducing agents. The purpose of the study was to evaluate the effects of ...ultraviolet (UV) radiation on the viability and the type of cell death on the human endometrial stromal cells (ThESC) line.
We investigated the effect of UV exposure on human endometrial stromal cell line (ThESC) on cell viability using MTT assay as well as changes in cell morphology using phase microscopy and acridine orange (AO)/ethidium bromide (EB) cell staining.
UV treatment significantly decreased the percentage of the viable ThESC cells compared to the viability of untreated control cells using MTT assay (p<0.05). In addition, UV treatment of ThESC cells for 60 and 90 min induced high level of cell morphology disruption, followed with loss of both the cell shape and the presence of defragmented debris and stained with intense red color.
The obtained results suggest the potential role of UV light application as additional treatment option of benign endometrium hyperplasia alone or in combination with other treatment modalities.
Background and purpose: Cytotoxic effects of Ligustrum vulgare leaves on HeLa cervical tumor cells suggested that Ligustrum vulgare extracts should be investigated as potential anticancer agents. ...Therefore, we examined a potential antileukemic activity of methanolic extracts of Ligustrum vulgare leaves and fruit extracts on two types of leukemia cells, MOLT-4 and JVM-13, lymphocytes isolated from the blood of 33 chronic lymphocytic leukemia (CLL) patients and on mononuclear leukocytes isolated from the blood of 18 healthy individuals.Material and methods: The cytotoxicity of examined extracts was measured by MTT assay and LDH activity test. The antiapoptotic potential of tested extracts was measured by Annexin V/7AAD flowcytometric assay.Results: The results showed that both extracts exhibited a moderate cytotoxic effect on all three types of leukemia cells. The Ligustrum vulgare leaf extract was the most effective on MOLT-4 cells, the fruit extract on JVM-13 cells and both extracts were equally effective on CLL cells. In addition, none of the tested extracts was toxic to healthy mononuclear cells. Both extracts acted by inducing apoptosis of leukemic cells.Conclusion: Ligustrum vulgare extracts exhibit significant antileukemic potential and should be further investigated.
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