细胞技术

Analysis of gene expression within tumors is frequently performed on the messenger RNA (mRNA). This chapter first describes the purification of mRNA from either tissues or cell culture and then describes the quantification by ribonuclease protection assay. Having initially purif ...

For cancer cells to form a metastasis, cells from the primary tumor must overcome the local adhesive forces, migrate and invade the microcirculation, arrest at a secondary site, and then finally proliferate (1). As implied by its multstep nature, cancer metastasis is a complex and dynamic proce ...

The application of Southern blotting to determine the methylation status of a particular gene has already been alluded to in Chapter 17 by Tennant et al., and methodology for Southern blotting described. This chapter examines methylation analysis of CpG islands in more depth and describes a t ...

In oncology, the correct assessment of tumor stage is crucial information, as it relates to disease-free interval and prognosis. Routine staging modalities use histopathology to evaluate the presence or absence of metastasis mainly in tissues such as lymph nodes, as described in Chapter 1 ...

Mathematical modeling is emerging as a powerful predictive tool in many areas of biology and medicine, with applications to cancer metastasis increasingly widespread and effective. This type of modeling involves quantitatively accurate representations of specific cellular ...

Cell-free systems derived from Xenopus eggs represent a powerful tool, intermediate between in vitro and in vivo model systems. Here, we describe protocols to prepare cell-free extracts recapitulating several aspects of the DNA damage response, including the DNA damage-dependent a ...

A short-term whole-skin organ culture model has been established to enable the investigation of cell cycle perturbations in epidermal layer cells following exposure to ultraviolet radiation (UVR). This model affords the opportunity to manipulate the growth and nutrient conditio ...

The checkpoint kinase Chk2 is activated in response to DNA damage through pathways requiring protein kinases ATM and/or ATR. The means by which Chk2 is activated by these kinases still remains to be addressed. Here we describe a cell-free system to study the activation of Chk2. Chk2 produced by a wheat ...

The clinical use of tumor necrosis factor (TNF) as an anticancer drug has been so far limited to loco-regional treatments because of severe dose-limiting toxicity. This chapter intends to review the targeting approaches and the animal models that have been developed in an attempt to improve the ...

To study tumor therapeutic treatment modalities, whether from a clinical, preclinical, or fundamental point of view, the use of clinically relevant animal models is indispensable. Particularly when the treatment comprises a multitargeted approach, (e.g., both tumor cells and endo ...

Among the cardiovascular pathologies, ischemic heart disease is the leading cause of congestive heart failure as well as permanent premature disabilities. Reperfusion of a previously ischemic heart is a standard clinical procedure. Even if beneficial, reperfusion triggers an in ...

Endotoxin/lipopolysaccharide (LPS) is the major mediator that triggers the cellular and humoral responses of the shock induced by Gram-negative bacteria. The toxic responses of LPS are mediated by various factors and mainly by tumor necrosis factor α (TNFα). To study the role of TNF and to iden ...

This chapter describes the methods for measuring the increase in vascular permeability induced by tumor necrosis factor α (TNFα), or other mediators, in vivo in animal models. Mouse liver or kidney are perfused through the portal vein or the renal artery, respectively, by intravascular inje ...

Tumor necrosis factor α (TNFα) is a crucial mediator involved in the communications between immune and nervous systems in physiological conditions, and its relevance is amplified during disease. Considered originally detrimental and a target for therapeutic intervention, rece ...

Tumor necrosis factor (TNF) is a pleiotropic cytokine that signals inflammation as well as cell death. We focus herein on the inflammatory pathway, giving particular emphasis to the in vitro methods used to study intracellular signaling mediators. The signal transduction cascade that T ...

We describe a semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) technique for the quantification of the messenger RNA of human and murine tumor necrosis factor α (TNFα) and related receptors. This protocol can be adapted for blood, peripheral blood lymphoc ...

Central to the tumor suppressor activity of certain proteins is the ability to interact physically with DNA. A well-studied example of this is the tumor suppressor p53 (1,2). The p53 protein has been implicated in several diverse growth-related pathways, including apoptosis and cell cycle a ...

The field of diagnostics has gathered the first crop of the genetic insurgency in the identification of the genetic basis of common and uncommon hereditary diseases. Discoveries of genes that predispose to cancer have led to the development of genetic tests. In turn, genetic tests can provide i ...

The classic paper by Schmitz and Galas (1) established the usefulness of footprinting analysis for identifying protein-bound sites on DNA. The basis of the footprinting technique is that DNA-bound proteins protect the phosphodiester backbone of DNA from modification or cleavage by ex ...

Gene expression is controlled at several steps. Transcription is one of the important steps at which regulation occurs. Tumor suppressors such as p53, which are basically transcription factors, carry out their function primarily by the transcriptional activation of the target genes. ...