TLR family gene expression in relation to the HIF1α and the VEGFR pathway activation in endometrial cancer

Introduction: Malignant neoplasm of the endometrium is the most common malignant neoplasm of the female reproductive system. Toll Like Receptors (TLR) play a significant role in innate and late-immunity against infections or damaged tissues. TLRs are also involved in the development of tumors in their natural microenvironment. TLRs play an important role in angiogenesis, necessary for survival and growth of the tumor. Hypoxia playing a critical role in angiogenesis, carcinogenesis, tumor progression and distant metastasis is primarily mediated through hypoxia inducible factors (HIFs). Vascular endothelial growth factor family proteins (VEGF) are also strongly involved in tumor angiogenesis and their action is strongly associated with TLR receptors.Objectives: The aim of the study was to correlate the expression of selected TLRs and VEGFR’s as well as HIF1α with clinicopathological data of endometrial cancer patients. Material and methods: 123 neoplastic endometrial samples were included in the study. 51 samples of healthy endometrium served as control. The expression of TLR1, TLR2, TLR3, TLR4, VEGFR1 and VEGFR2, VEGF-A and HIF1α was examined after RNA isolation at the mRNA level by Real Time-PCR.Results: We have noted a significant correlation between the expression of selected TLR and VEGFR’s and clinical stage as well as pathological grading of endometrial cancer.Conclusions: Received correlations confirm a significant contribution of some TLR expression and the receptor for VEGF in the pathogenesis of epithelial endometrial cancer

Expression of TopBP1 in hereditary breast cancer

TopBP1 protein displays structural as well as functional similarities to BRCA1 and is involved in DNA replication, DNA damage checkpoint response and transcriptional regulation. Aberrant expression of TopBP1 may lead to genomic instability and can have pathological consequences. In this study we aimed to investigate expression of TopBP1 gene at mRNA and protein level in hereditary breast cancer. Real-time quantitative PCR was performed in 127 breast cancer samples. Expression of TopBP1 mRNA in lobular carcinoma was significantly lower compared with ductal carcinoma (p < 0.05). The level of TopBP1 mRNA appeared to be lower in poorly differentiated (III grade) hereditary breast cancer in comparison with moderately (II grade) and well-differentiated cancer (I grade) (p < 0.05 and p < 0.001 respectively). We analyzed TopBP1 protein expression using immunohistochemistry and Western blot techniques. Expression of TopBP1 protein was found to be significantly increased in poorly differentiated breast cancer (III grade) (p < 0.05). The percentage of samples with cytoplasmic apart from nuclear staining increased with increasing histological grade. There was no significant association between level and intracellular localization of TopBP1 protein in hereditary breast cancer and other clinicopathological parameters such as estrogen and progesterone receptors status, appearance of metastasis in the axillary lymph nodes and type of cancer. Our data suggest that decreased level of TopBP1 mRNA and increased level of TopBP1 protein might be associated with progression of hereditary breast cancer

Relationship of urinary isoprostanes to prostate cancer occurence

To estimate the oxidative stress in patients with prostate cancer and in a control group, we used the biomarker of lipid peroxidation–isoprostanes (8-isoPGF2) and the level of selected antioxidants (glucose and uric acid [UA]). The level of urinary isoprostanes was determined in patients and controls using an immunoassay kit according to the manufacturer’s instruction. The levels of UA and glucose were also determined in serum by the use of UA Assay Kit and Glucose Assay Kit. We observed a statistically increased the level of isoprostanes in urine of patients with prostate cancer in compared with a control group. The concentration of tested antioxidants in blood from patients with prostate cancer was also higher than in healthy subjects. Moreover, our experiments indicate that the correlation between the increased amount of UA and the lipid peroxidation exists in prostate cancer patients (in all tested groups). Prostate cancer risk by urinary isoprostanes level was analyzed, and a positive association was found (relative risk for highest vs. lowest quartile of urinary isoprostanes = 1.6; 95 % confidence interval 1.2–2.4; p for trend = 0.03). We suggest that reactive oxygen species induce peroxidation of unsaturated fatty acid in patients with prostate cancer, and the level of isoprostanes may be used as a non-invasive marker for determination of oxidative stress. We also propose that UA may enhance the oxidative stress in patients with prostate cancer.This study was supported by the Grant 506/810(KBO) from University of Lodz, Polan

Trypanosoma brucei infection elicits nitric oxide-dependent and nitric oxide-independent suppressive mechanisms

During murine Trypanosoma brucei infection, macrophages contribute significantly to the inhibition of T cell responses, Although nitric oxide (NO) was shown to play a central role in macrophage-mediated splenic suppression, macrophage-mediated lymph node suppression occurred in an interferon-gamma (IFN-gamma)-dependent manner, In this study, using NO inhibitor N-G-monomethyl-L-arginine and anti-IFN-gamma antibodies, the relative contribution of NO and IFN-gamma to the active inhibition of ex vivo concanavalin A-induced T cell proliferation taking place in the spleen and the lymph nodes of T, brucei-infected mice was investigated, NO contributes to the suppressive activity of spleen and lymph node cells only during early-stage infection, The existence of NO-independent suppressive pathway was further evidenced in IFN-gamma(-/-)-infected mice, Spleen cells from such animals do not produce NO but exert significant suppressive activity during the whole course of infection, In contrast in the lymph nodes, no suppressive activity is recorded at any moment of infection, Moreover, addition of exogenous IFN-gamma to cultures containing lymph node cells from IFN-gamma(-/-)-infected mice does not impair proliferation despite NO production in such cultures, Thus during late-stage infection, an IFN-gamma-independent suppressive mechanism is elicited in the spleen, whereas in the lymph nodes, IFN-gamma is required yet not sufficient to inhibit T cell proliferation

Trypanosoma brucei infection elicits nitric oxide-dependent and nitric oxide-independent suppressive mechanisms

During murine Trypanosoma brucei infection, macrophages contribute significantly to the inhibition of T cell responses, Although nitric oxide (NO) was shown to play a central role in macrophage-mediated splenic suppression, macrophage-mediated lymph node suppression occurred in an interferon-gamma (IFN-gamma)-dependent manner, In this study, using NO inhibitor N-G-monomethyl-L-arginine and anti-IFN-gamma antibodies, the relative contribution of NO and IFN-gamma to the active inhibition of ex vivo concanavalin A-induced T cell proliferation taking place in the spleen and the lymph nodes of T, brucei-infected mice was investigated, NO contributes to the suppressive activity of spleen and lymph node cells only during early-stage infection, The existence of NO-independent suppressive pathway was further evidenced in IFN-gamma(-/-)-infected mice, Spleen cells from such animals do not produce NO but exert significant suppressive activity during the whole course of infection, In contrast in the lymph nodes, no suppressive activity is recorded at any moment of infection, Moreover, addition of exogenous IFN-gamma to cultures containing lymph node cells from IFN-gamma(-/-)-infected mice does not impair proliferation despite NO production in such cultures, Thus during late-stage infection, an IFN-gamma-independent suppressive mechanism is elicited in the spleen, whereas in the lymph nodes, IFN-gamma is required yet not sufficient to inhibit T cell proliferation