XPD codon 312 and 751 polymorphisms, and AFB1 exposure, and hepatocellular carcinoma risk

<p>Abstract</p> <p>Background</p> <p>Genetic polymorphisms in DNA repair genes may influence individual variation in DNA repair capacity, which may be associated with risk of hepatocellular carcinoma (HCC) related to the exposure of aflatoxin B1 (AFB1). In this study, we have focused on the polymorphisms of xeroderma pigmentosum complementation group D (XPD) codon 312 and 751 (namely Asp312Asn and Lys751Gln), involved in nucleotide excision repair.</p> <p>Methods</p> <p>We conducted a case-control study including 618 HCC cases and 712 controls to evaluate the associations between these two polymorphisms and HCC risk for Guangxi population by means of TaqMan-PCR and PCR-RFLP analysis.</p> <p>Results</p> <p>We found that individuals featuring the XPD genotypes with codon 751 Gln alleles (namely XPD-LG or XPD-GG) were related to an elevated risk of HCC compared to those with the homozygote of XPD codon 751 Lys alleles [namely XPD-LL, adjusted odds ratios (ORs) were 1.75 and 2.47; 95% confidence interval (CIs) were 1.30-2.37 and 1.62-3.76, respectively]. A gender-specific role was evident that showed an higher risk for women (adjusted OR was 8.58 for XPD-GG) than for men (adjusted OR = 2.90 for XPD-GG). Interestingly, the interactive effects of this polymorphism and AFB1-exposure information showed the codon 751 Gln alleles increase the risk of HCC for individuals facing longer exposure years (<it>P</it>_interaction= 0.011, OR = 0.85). For example, long-exposure-years (> 48 years) individuals who carried XDP-GG had an adjusted OR of 470.25, whereas long-exposure-years people with XDP-LL were at lower risk (adjusted OR = 149.12). However, we did not find that XPD codon 312 polymorphism was significantly associated with HCC risk.</p> <p>Conclusion</p> <p>These findings suggest that XPD Lys751Gln polymorphism is an important modulator of AFB1 related-HCC development in Guangxi population.</p

STAT1-dependent expression of energy metabolic pathways links tumour growth and radioresistance to the Warburg effect

<p>Abstract</p> <p>Background</p> <p>The Signal Transducer and Activator of Transcription 1 (STAT1) has traditionally been regarded as a transmitter of interferon signaling and a pro-apoptotic tumour suppressor. Recent data have identified new functions of STAT1 associated with tumourigenesis and resistance to genotoxic stress, including ionizing radiation (IR) and chemotherapy. To investigate the mechanisms contributing to the tumourigenic functions of STAT1, we performed a combined transcriptomic-proteomic expressional analysis and found that STAT1 is associated with regulation of energy metabolism with potential implication in the Warburg effect.</p> <p>Methods</p> <p>We generated a stable knockdown of STAT1 in the SCC61 human squamous cell carcinoma cell line, established tumour xenografts in athymic mice, and compared transcriptomic and proteomic profiles of STAT1 wild-type (WT) and knockdown (KD) untreated or irradiated (IR) tumours. Transcriptional profiling was based on Affymetrix Human GeneChip^®Gene 1.0 ST microarrays. Proteomes were determined from the tandem mass spectrometry (MS/MS) data by searching against the human subset of the UniProt database. Data were analysed using Significance Analysis of Microarrays for ribonucleic acid and Visualize software for proteins. Functional analysis was performed with Ingenuity Pathway Analysis with statistical significance measured by Fisher's exact test.</p> <p>Results</p> <p>Knockdown of STAT1 led to significant growth suppression in untreated tumours and radio sensitization of irradiated tumours. These changes were accompanied by alterations in the expression of genes and proteins of glycolysis/gluconeogenesis (GG), the citrate cycle (CC) and oxidative phosphorylation (OP). Of these pathways, GG had the most concordant changes in gene and protein expression and demonstrated a STAT1-dependent expression of genes and proteins consistent with tumour-specific glycolysis. In addition, IR drastically suppressed the GG pathway in STAT1 KD tumours without significant change in STAT1 WT tumours.</p> <p>Conclusion</p> <p>Our results identify a previously uncharacterized function of STAT1 in tumours: expressional regulation of genes encoding proteins involved in glycolysis, the citrate cycle and mitochondrial oxidative phosphorylation, with predominant regulation of glycolytic genes. STAT1-dependent expressional regulation of glycolysis suggests a potential role for STAT1 as a transcriptional modulator of genes responsible for the Warburg effect.</p

Radiation-Induced Bystander Effects in Cultured Human Stem Cells

The radiation-induced "bystander effect" (RIBE) was shown to occur in a number of experimental systems both in vitro and in vivo as a result of exposure to ionizing radiation (IR). RIBE manifests itself by intercellular communication from irradiated cells to non-irradiated cells which may cause DNA damage and eventual death in these bystander cells. It is known that human stem cells (hSC) are ultimately involved in numerous crucial biological processes such as embryologic development; maintenance of normal homeostasis; aging; and aging-related pathologies such as cancerogenesis and other diseases. However, very little is known about radiation-induced bystander effect in hSC. To mechanistically interrogate RIBE responses and to gain novel insights into RIBE specifically in hSC compartment, both medium transfer and cell co-culture bystander protocols were employed.Human bone-marrow mesenchymal stem cells (hMSC) and embryonic stem cells (hESC) were irradiated with doses 0.2 Gy, 2 Gy and 10 Gy of X-rays, allowed to recover either for 1 hr or 24 hr. Then conditioned medium was collected and transferred to non-irradiated hSC for time course studies. In addition, irradiated hMSC were labeled with a vital CMRA dye and co-cultured with non-irradiated bystander hMSC. The medium transfer data showed no evidence for RIBE either in hMSC and hESC by the criteria of induction of DNA damage and for apoptotic cell death compared to non-irradiated cells (p>0.05). A lack of robust RIBE was also demonstrated in hMSC co-cultured with irradiated cells (p>0.05).These data indicate that hSC might not be susceptible to damaging effects of RIBE signaling compared to differentiated adult human somatic cells as shown previously. This finding could have profound implications in a field of radiation biology/oncology, in evaluating radiation risk of IR exposures, and for the safety and efficacy of hSC regenerative-based therapies

Position weight matrix model as a tool for the study of regulatory elements distribution across the DNA sequence

Ab initio methods of DNA regulatory sequence region prediction known as transcription factor binding sites (TFBS) are a very big challenge to modern bioinformatics. Although the currently available methods are not perfect they are fairly reliable and can be used to search for new potential protein-DNA interaction sites. The biggest problem of ab initio approaches is the very high false positive rate of predicted sites which results mainly from the fact that TFBS are very short and highly degenerate. Because of that they can occur by chance every few hundred bases making the task of computational prediction extremely difficult if one aims to reduce the high false positive rate keeping highest possible sensitivity to predict biologically meaningful sequence regions. In this work we present a new application that can be used to predict TFBS regions in very large datasets based on position weight matrix models (PWM’s) using one of the most popular prediction methods. The presented application was used to predict the concentration of TFBS in a set of nearly 2.2 thousand unique sequences of human gene promoter regions. The study revealed that the concentration of TFBS further than 1kbp from the transcription initiation site is constant but it decreases rapidly while getting closer to the transcription initiation site. The decreasing TFBS concentration in the vicinity of genes might result from evolutionary selection which keeps only sites responsible for interactions with proteins being part of a specific regulatory mechanism leading to cells survival

Are infinite dimensional models applicable in modelling and analysis of cancer chemotherapy?

Drug resistance and phase dependence have been regarded by many authors as the main obstacles against successful cancer chemotherapy. We propose a model which takes into account both these phenomena and give a tool to use phase specificity as an advantage rather than a fault and make it resistant of drug resistance. It combines models that so far have been studied separately, taking into account both the phenomenon of gene amplification and drug specificity in chemotherapy, in their different aspects. The mathematical description is given by an infinite dimensional state equation with a system matrix, the form of which enables decomposition of the model into two interacting subsystems. While the first one, of finite dimension, can have any form, the second one is infinite dimensional and tridiagonal

Complex of DNA with chromatin proteins investigated by isopycnic centrifugation in metrizamide.

Complexes of mouse main band DNA with a fraction of non-histone proteins (NHP), having a high affinity for DNA, in the absence or presence of histones have been investigated by gradient centrifugation in metrizamide. Two types of complexes were formed at an input ratio of NHP to DNA between 1 and 2.5. In metrizamide gradients a majority of DNA was found in the light complex (at the density of 1.14-1.16 g/cm3) even at the very high NHP to DNA ratio. When histones were present in the reaction mixture, most of the DNA was found in the heavy complex (1.19-1.21 g/cm3). The electrophoretic profiles of the proteins recovered from the heavy and light complexes were different; some fractions of nonhistone proteins were present only in the heavy component

Analiza asymptotyczna trzech modeli błądzenia z rozgałęzieniami z dziedziny biologii molekularnej

Using asymptotic techniques based on Laplace transforms, spectral analysis and theory of feedback systems, we characterise the asymptotic behaviour of the repeat loci in microsatellite DNA and cancer cells with increasing number of copies of genes responsible for coding proteins causing drug removal or metabolisation as well as telomeres shortening, which is supposed to be the mechanism of ageing and death. These three problems are described by models in the form of infinitely many differential linear or bilinear first order equations, resulting from branching random walk processes used to represent the evolution of particles in these problems.Wykorzystując techniki asymptotyczne oparte na transformatach Laplace'a, analizę spektralną oraz teorię układów ze sprzężeniem zwrotnym w artykule scharakteryzowano zachowanie asymptotyczne powtórek w DNA mikrosatelitarnym oraz w komórkach rakowych z rosnącą liczbą kopii genów odpowiedzialnych za kodowanie białek powodujących usuwanie lub przemianę metaboliczną leków, a także skracanie telomerów, o którym się sądzi, że jest mechanizmem starzenia się i śmierci. Te trzy zagadnienia są opisywane przy pomocy modeli w postaci nieskończonej liczby liniowych lub biliniowych równań pierwszego rzędu, wynikających z procesów błądzenia, stosowanych do opisu ewolucji cząstek w tych zagadnieniach

Crosstalk between transcription factors in regulation of the human glutathione S-transferase P1 gene expression in Me45 melanoma cells

Aim. The human GSTP1 is a major enzyme of phase II detoxification in the most cell types. Aberrant expression of GSTP1 is associated with carcinogenesis and development of multidrug resistance. The GSTP1 gene expression is regulated at multiple levels including transcriptional, post-transcriptional and post-translational. We concentrated our attention on the transcriptional level of regulation. Methods. Transient transfection of Me45 melanoma cells with constructs containing the luciferase gene under the control of complete and truncated GSTP1 promoter was utilized to identify a role of different promoter regions in regulation of the gene transcription in Me45 cells. To identify the transcription factors, interacting with the GSTP1 promoter sites, the competitive EMSA and super shift assay were applied. Results. GSTP1 transcription in Me45 cells is positively regulated by binding NF-κB to the cognate site and ERβin complex with unknown protein to the ARE site; the complex of ERβ with c-Fos negatively regulates the gene expression via CRE site. The interaction of c-Fos/ERβ with GSTP1 CRE site and indirect interaction of ERβ with GSTP1 ARE were identified. Conclusions. The positive regulation of the human GSTP1 gene in Me45 melanoma cells is exerted via NF-κB and ARE sites and the negative one via CRE site of the promoter. ERis indirectly involved in the regulation of GSTP1 transcription. It is bound via c-Fos with CRE site and via unknown protein with ARE site

Transcription regulation in differential expression of the human GSTP1 gene in breast and choriocarcinoma cells

Glutathione S-transferase P1 is a major phase II detoxification enzyme in most cell types. Aberrant expression of GSTP1 is associated with carcinogenesis and development of multidrug resistance. GSTP1 gene transcription is regulated by promoter methylation and by transcription factors. To elucidate the mechanisms responsible for the different levels of GSTP1 expression observed in Hbl-100 and BeWo cells we utilized truncated promoter constructs to compare the functional role of different promoter elements. We also identified transcription factors binding the responsive elements by electrophoretic mobility shift assay. The applied approaches provided the evidence that binding of transcription factors to ARE, CRE and NF-kappaB sites are responsible for the cell specific levels of GSTP1 expression in Hbl-100 and BeWo cells. It was also indicated that partial promoter methylation occurs in BeWo cells