Regulation of the cell fate by DNA damage and hypoxia

In order to provide the means for the design of novel rational anti-cancer drug therapies research efforts are concentrated on unravelling the molecular circuits which induce programmed cell death and block proliferation of cancer cells.Modern therapeutic strategies are based on the understanding of the complexity of physiological functions such as differentiation,development,immune responses,cell-cycle arrest,DNA damage repair,apoptosis,autophagy,energy metabolism,and senescence.It has become evident that this knowledge will provide the means to target the components of the pathways involved in these processes in a specific and selective manner thus paving the way for the development of effective and personalised anti-cancer therapies.Transcription is a crucial cellular process that regulates a multitude of physiological functions,which are essential in disease progression and cellular response to therapy.Transcription factors such as the p53 tumor suppressor and the hypoxia-inducible factor-α(HIF-α) are key players in carcinogenesis and cellular response to cancer therapies.Both of these transcription factors regulate gene expression of genes involved in cell death and proliferation,in some cases cooperating towards producing the same outcome and in some others mediating opposing effects.It is thus apparent that fine tuning of the activity of these transcription factors is essential to determine the cellular response to therapeutic regimens,in other words whether tumor cells will commit to apoptosis or evade engagement with the anti-proliferative effects of drugs leading to drug resistance.Our observations support the notion that the functional crosstalk between HIF-1α and p53 pathways and thus the fine tuning of their transcriptional activity is mediated by cofactors shared between the two transcription factors such as components of the p300 co-activator multiprotein complex.In particular,there is evidence to suggest that differential composition of the co-modulatory protein complexes associated with p53 and HIF-la under diverse types of stress conditions differentially regulate the expression of distinct subsets of p53 and HIF-la target genes involved in processes such as cell cycle arrest,apoptosis,chronic inflammation,and cellular energy metabolism thereby determining the cellular fate under particular types of microenvironmental stress.

Oxidative stress and breast cancer biomarkers:the case of the cytochrome P450 2E1

Aim:The aim of the study is to investigate the impact of the cytochrome P4502E1,which is the most efficient CYP450 family member in generating reactive oxygen species(ROS),on cellular energy metabolism of breast cancer cells and therefore the effects of CYP2E1 on breast carcinogenesis.Methods:The estrogen receptor positive MCF-7 and the triple negative MDA-MB-231 breast cancer cells were used as experimental system to estimate ROS generation in these cells overexpressing CYP2E1 and treated with the glycolytic inhibitors 3-bromopyruvate or 2-deoxyglucose in the presence or absence of the CYP2E1 inhibitor chlormethiazole.Adenosine triphosphate(ATP)assay was used to measure ATP production and lactate assay to quantify the efflux of lactic acid in breast cancer cells treated with the CYP2E1 inhibitor chlormethiazole,the mitochondrial membrane potential and cell viability assays were employed to assess the pathway of cellular energy production and cellular death respectively after treatment of MCF-7 and MDA-MB-231 with the CYP2E1 activator acetaminophen or the CYP2E1 inhibitor chlormethiazole.Results:T he r esults i ndicated i ncreased ROS generation i n b reast c ancer c ells overexpressing C YP2E1.ROS generation was differentially regulated in breast cancer cells upon treatment with the CYP2E1 inhibitor chlormethiazole.Chlormethiazole treated MCF-7 cells exhibited reduced lactate efflux implying that CYP2E1 directly or indirectly regulates the glycolytic rate in these cells.Furthermore the mitochondrial membrane potential of both MCF-7 and MDA-MB-231 cells was differentially affected by the CYP2E1 activator acetaminophen versus the CYP2E1 inhibitor chlormethiazole providing additional support for the involvement of CYP2E1 in energy metabolic pathways in breast cancer.Conclusion:Results presented in this study provide evidence to suggest that CYP2E1 regulates cellular energy metabolism of breast cancer cells in a manner dependent on cell type and potentially on the clinical staging of the disease therefore CYP2E1 is a possible breast cancer biomarker.