Springer protocols feed by Cancer Research 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Application of RNA-Seq Technology in Cancer Chemoprevention
RNA-sequencing is a revolutionary tool to follow differential expression after treatment with cancer chemopreventive agents. It allows a real genome-wide screening independent of prior assumptions and is well suited for analyzing coding but also long noncoding RNAs. It still consents the discovery of new genes and isoforms and increased our knowledge of antisense and other noncoding RNAs in a tremendous manner. Moreover, it permits to detect low-abundance and biologically critical isoforms and reveals genetic variants and gene fusions in one single assay. Here, we provide a detailed protocol for stranded RNA-sequencing. (S...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
microRNAs in Cancer Chemoprevention: Method to Isolate Them from Fresh Tissues
microRNAs are 22-nucleotide-long double-strand small RNAs, able to modulate gene expression at posttranscriptional level, degrading mRNA and/or impairing translation. They have been shown to regulate mRNA and protein abundance and to participate in many regulatory circuits controlling developmental timing, cell proliferation and differentiation, apoptosis and stress response. Notably, microRNA activity has been correlated to the pathogenesis of cancer; they are aberrantly expressed in solid and hematological tumors, suggesting that they could function as oncogenes or tumor suppressors. The emerging role of miRNAs in the ca...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Use of Buffy Coat miRNA Profiling for Breast Cancer Prediction in Healthy Women
MicroRNAs are key regulators of different biological processes and their deregulation is associated with the occurrence of many diseases among which cancer. Due to the higher stability of microRNAs and to the easiness in their detection both in organs than in biological fluids, many studies are turned toward potential use of this small molecules as biomarkers for the prediction and diagnosis of different types of cancer. Here we describe the experiment protocol that we have used for microRNA profiling analysis in buffy coat samples of women who developed breast cancer versus women who remained healthy during a 20 year foll...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Controlled Delivery of Chemopreventive Agents by Polymeric Implants
The clinical development of cancer chemopreventive agents has been hampered by poor oral bioavailability issue. Several compounds have low aqueous solubility and undergo extensive first pass metabolism following oral dosing. To overcome this limitation, we developed polymeric implants from biodegradable ε-polycaprolactone (PCL) that can deliver both lipophilic as well as hydrophilic compounds. Implants furnish controlled release of compounds for long duration and provide dose-dependent release. The rate of release in vitro correlated well with the in vivo release. The polymeric implant technology thus overcomes the...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
South African Herbal Extracts as Potential Chemopreventive Agents: Screening for Anticancer Splicing Activity
RT-PCR is an invaluable tool for the detection and characterization of mRNA. Cancer cell lines are treated with crude plant extracts and RNA is extracted and purified with DNase prior to RT-PCR. RT-PCR first-strand cDNA synthesis is done using random primers and can be refrigerated at 4 °C. PCR from the stored cDNA is performed using transcript-specific primers and electrophoresed on a molecular grade agarose gel to separate the splice variants. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Antibody Array as a Tool for Screening of Natural Agents in Cancer Chemoprevention
The efficacy of a given drug resides mainly on its ability to specifically target disease mechanisms. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
3D Tumor Models and Time-Lapse Analysis by Multidimensional Microscopy
The 3D culture is advantageous in reflecting the in vivo condition compared to the 2D culture; however, imaging 3D-cultured cells may be a challenge due to technical restrictions. Recent development of confocal spinning disc microscope system as well as sophisticated software has enabled us to monitor dynamism of cell movement in multiple dimensions. Here we describe the method for time-lapse imaging of 3D-cultured cancer cells co-cultured with non-cancerous cells and discuss current limitations and future perspectives. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Protocol for a Steady-State FRET Assay in Cancer Chemoprevention
Cancer chemoprevention is an important strategy to prevent, reverse, or suppress the development of cancer. One of the target pathways that has emerged in recent years is the Keap1-Nrf2-ARE system that regulates the protection of cells against various carcinogens and their metabolites. Increased concentrations of the redox transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) induces the activation of antioxidant and phase 2 detoxifying genes. Nrf2 is regulated by substrate adaptor protein Kelch-like ECH-associated protein 1 (Keap1) that can target Nrf2 for ubiquitination and degradation by the proteasome...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Autophagy in Cancer Chemoprevention: Identification of Novel Autophagy Modulators with Anticancer Potential
Cancer cells have the ability to tolerate extreme conditions, autophagy-related stress tolerance enables cancer cells to survive by maintaining energy production that leads to cell growth and therapeutic resistance. Insufficient activation of autophagy in nutrient-deprived cancer cells may sensitize cancer cells to a broad array of chemotherapeutic agents and ionizing radiation. Therefore, identification of novel autophagy modulators with lower toxicity and better therapeutic index would be beneficial for cancer therapy. Here, we describe several currently used biochemical methods to assess autophagic activity and lysosoma...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Isolation of Chemoresistant Cell Subpopulations
Chemoresistance is a major challenge for cancer therapy and drives tumor relapse. The emergence, within the treated tumor mass, of specific cancer cell subpopulations endowed with high tolerance to the microenvironment stress induced by therapy is being growingly recognized as a mechanism of tumor progression. To obtain detailed information with regard to the pathways underlying survival, expansion, and microenvironmental cross talk of such chemoresistant cell subpopulations may be instrumental for cancer chemoprevention. Additionally, the obtained cell subpopulations may be used for direct screening of cancer chemoprevent...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Cancer Gastric Chemoprevention: Isolation of Gastric Tumor-Initiating Cells
Gastric cancer is an important healthcare problem and represents the second leading cause of death for malignant disease worldwide. In the Western world, the diagnosis is done at late stage when treatments can be only palliative. Searches for new therapeutic regimens as well as for new biomarkers are in progress. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
AlgiMatrix™-Based 3D Cell Culture System as an In Vitro Tumor Model: An Important Tool in Cancer Research
Routinely used two-dimensional cell culture-based models often fail while translating the observations into in vivo models. This setback is more common in cancer research, due to several reasons. The extracellular matrix and cell-to-cell interactions are not present in two-dimensional (2D) cell culture models. Diffusion of drug molecules into cancer cells is hindered by barriers of extracellular components in in vivo conditions, these barriers are absent in 2D cell culture models. To better mimic or simulate the in vivo conditions present in tumors, the current study used the alginate based three-dimensional cell culture (...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Angiogenesis Assays
Neoangiogenesis constitutes one of the first steps of tumor progression beyond a critical size of tumor growth, which supplies a dormant mass of cancerous cells with the required nutrient supply and gaseous exchange through blood vessels essentially needed for their sustained and aggressive growth. In order to understand any biological process, it becomes imperative that we use models, which could mimic the actual biological system as closely as possible. Hence, finding the most appropriate model is always a vital part of any experimental design. Angiogenesis research has also been much affected due to lack of simple, reli...
Source: Springer protocols feed by Cancer Research - November 28, 2015 Category: Cancer & Oncology Source Type: news
Methods for Assessing the In Vivo Role of PTEN in Glucose Homeostasis
We describe the generation of tissue-specific PTEN knockout mice and models of both type 1 and type 2 diabetes, which we have found useful for the study of diabetes pathogenesis. We also outline common methods suitable for the characterization of glucose homeostasis in rodent models, including techniques to measure beta cell function and insulin sensitivity. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 18, 2015 Category: Cancer & Oncology Source Type: news
Germline PTEN Mutation Analysis for PTEN Hamartoma Tumor Syndrome
Clinically, deregulation of PTEN function resulting in reduced PTEN expression and/or activity is implicated in human disease. Cowden syndrome (CS) is an autosomal dominant disorder characterized by benign and malignant tumors. CS-related individual features occur commonly in the general population. Approximately 25 % of patients diagnosed with CS have pathogenic germline PTEN mutations, which increase lifetime risks of breast, thyroid, uterine, renal, and other cancers. PTEN testing and intensive cancer surveillance allow for early detection and treatment of these cancers for mutation-positive patients and their relatives...
Source: Springer protocols feed by Cancer Research - November 18, 2015 Category: Cancer & Oncology Source Type: news
