Journal: Cancer Research

Article Title: Gene Expression Analysis of Preinvasive and Invasive Cervical Squamous Cell Carcinomas Identifies HOXC10 as a Key Mediator of Invasion

doi: 10.1158/0008-5472.can-07-2056

Figure Lengend Snippet: Figure 2. Validation of microarray data by qRT-PCR and immunohistochemistry. A, comparison of HOXC10, HOXC6, and APOC1 gene expression from microarray and qRT-PCR analyses. RNA for qRT- PCR were extracted from SCC (n = 19), HSIL (n = 5), and normal cervix (n = 8) samples isolated by LCM. For each gene, the mean relative expression (normalized to HPRT1) based on qRT-PCR analysis is plotted against the expression measure obtained from the microarray analysis. The Pearson’s correlation (r) and its significance (P) are given. B, immunohistochemical analysis of Ect2, FoxM1, Rfc4, and TK1 protein expression in representative normal cervix (NCx), HSIL, and SCC tissue samples. Tissue sections were immunostained with antibodies recognizing the indicated proteins. Original magnification, 200.

Article Snippet: Sections were incubated with rabbit polyclonal anti-ECT2 (1:500 dilution), anti-FOXM1 (1:300 dilution), or mouse monoclonal anti-RCF4 (1:300 dilution) antibody (Santa Cruz Biotechnology, Inc.) or anti-TK1 antibody (1:200 dilution, Novus Biologicals, Inc.) overnight at 4jC.

Techniques: Biomarker Discovery, Microarray, Quantitative RT-PCR, Immunohistochemistry, Comparison, Gene Expression, Isolation, Expressing, Immunohistochemical staining

Journal: Biochemical pharmacology

Article Title: Accumulation of thymidine-derived sugars in thymidine phosphorylase overexpressing cells.

doi: 10.1016/j.bcp.2010.05.009

Figure Lengend Snippet: Fig. 1. Schematic overview of the conversion of TdR to TdR-related sugars and possible pathways which can stimulate angiogenesis. (A) TdR enters the cell, after which it is converted to thymine and deoxyribose-1-phosphate (dR-1-P). dR-1-P can be further converted to dR or dR-5-P. dR-5-P can enter the pentose phosphate pathway or glycolysis, stimulating the metabolism of the cells, possibly activating angiogenesis. dR is secreted by the cells, after which it can attract endothelial cells and/ or be taken up by the endothelial cells, stimulating angiogenesis. Both dR and dR-5-P can undergo Schiff base reactions, first by opening the ring structure of the sugar by anomerization, after which it can undergo a Schiff base reaction where an Amadori product is formed. Subsequently, reactive oxygen species (ROS) are formed, or advanced glycation end products (AGE). Moieties at which [50-3H]-TdR was labeled and the TdR-derived sugars are indicated with asterisk. TP: Thymidine phosphorylase; 1. thymidine kinase (TK); 2. DNA polymerase; 3. a phosphatase; 4. phospho-pentomutase; 6. deoxyribosephospatealdolase. (B) Structural formulae of the open ring structure of dR (2-deoxy-D-ribose), a Schiff base and an Amadori product. The open chain aldehyde glycates protein by a non-enzymatic condensation of the sugar to form a Schiff base, which rearranges to form an a- hydroxyketone Amadori product. dR-5-P can undergo the similar reaction. R = protein (see also ref. [13]).

Article Snippet: The thymidine phosphorylase antibody was purchased from R&D Systems Inc. (Minneapolis, MN, USA) and the thymidine kinase antibody was purchased from QED Bioscience (San Diego, CA, USA).

Techniques: Labeling, Derivative Assay

Journal: Biochemical pharmacology

Article Title: Accumulation of thymidine-derived sugars in thymidine phosphorylase overexpressing cells.

doi: 10.1016/j.bcp.2010.05.009

Figure Lengend Snippet: Fig. 2. Western blot showing the expression levels of thymidine phosphorylase in Colo320, Colo320 TP1, RT112 and RT112/TP cells.

Article Snippet: The thymidine phosphorylase antibody was purchased from R&D Systems Inc. (Minneapolis, MN, USA) and the thymidine kinase antibody was purchased from QED Bioscience (San Diego, CA, USA).

Techniques: Western Blot, Expressing

Journal: Biochemical pharmacology

Article Title: Accumulation of thymidine-derived sugars in thymidine phosphorylase overexpressing cells.

doi: 10.1016/j.bcp.2010.05.009

Figure Lengend Snippet: Fig. 3. The accumulation of sugars after conversion of TdR by thymidine phosphorylase. (A) Total levels of thymine and total levels of measured sugars. (B) Measurement of the levels of the various TdR-related sugars. (C) Measurement of the levels of dR that was secreted by the cells after degradation of TdR. All values represent means of three independent experiments SEM. Significant differences compared to the control levels are indicated *P < 0.05.

Article Snippet: The thymidine phosphorylase antibody was purchased from R&D Systems Inc. (Minneapolis, MN, USA) and the thymidine kinase antibody was purchased from QED Bioscience (San Diego, CA, USA).

Techniques: Control