scholarly journals Real-Time RT-PCR Analysis for Evaluating the Her2/neu Status in Breast Cancer

Pathobiology ◽  
2010 ◽  
Vol 77 (1) ◽  
pp. 38-45 ◽  
Author(s):  
M. Cuadros ◽  
P. Talavera ◽  
F.J. López ◽  
I. García-Peréz ◽  
A. Blanco ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Pcr Analysis ◽  
Rt Pcr

Quantitative real-time RT-PCR analysis of eight novel estrogen-regulated genes in breast cancer

2003 ◽  
Vol 18 (2) ◽  
pp. 123-129 ◽  
Author(s):  
V. Sorbello ◽  
L. Fuso ◽  
C. Sfiligoi ◽  
C. Scafoglio ◽  
R. Ponzone ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Pcr Analysis ◽  
Rt Pcr

scholarly journals Quantitative Real-Time RT-PCR Analysis of Eight Novel Estrogen-Regulated Genes in Breast Cancer

2003 ◽  
Vol 18 (2) ◽  
pp. 123-129 ◽  
Author(s):  
V. Sorbello ◽  
L. Fuso ◽  
C. Sfiligoi ◽  
C. Scafoglio ◽  
R. Ponzone ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Progesterone Receptors ◽  
Breast Tumors ◽  
Response To Treatment ◽  
Endocrine Treatment ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Variable Expression

Background Biological markers capable of predicting the risk of recurrence and the response to treatment in breast cancer are eagerly awaited. Estrogen and progesterone receptors (ER, PgR) in tumor cells mark cancers that are more likely to respond to endocrine treatment, but up to 40% of such patients do not respond. Here, the expression of a group of estrogen-regulated genes, previously identified by microarray analysis of in vitro models, was measured in breast tumors and possible associations with other clinicopathological variables were investigated. Methods The expression of CD24, CD44, HAT-1, BAK-1, G1P3, TIEG, NRP-1 and RXRα was measured by quantitative real-time RT-PCR on RNA from eighteen primary breast tumors. Statistical analyses were used to identify correlations among the eight genes and the available clinicopathological data. Results Variable expression levels of all the genes were observed in all the samples examined. Significant associations of CD24 with tumor size, CD44 with lymph node invasion, and HAT-1 and BAK-1 with ER positivity were found. The possible combinatorial value of these genes was assessed. Unsupervised hierarchical clustering analysis demonstrated that the expression profile of these genes was able to predict ER status with an acceptable approximation. Conclusions Eight novel potential markers for breast cancer have been preliminarily characterized. As expected from in vitro data, their expression is able to discriminate ER- versus ER+ tumors.

scholarly journals Reproductive failure in mice lacking inter-alpha-trypsin inhibitor (ITI) – ITI target genes in mouse ovary identified by microarray analysis

2004 ◽  
Vol 183 (1) ◽  
pp. 29-38 ◽  
Author(s):  
Mika Suzuki ◽  
Hiroshi Kobayashi ◽  
Yoshiko Tanaka ◽  
Naohiro Kanayama ◽  
Toshihiko Terao
Keyword(s):  
Real Time ◽  
Trypsin Inhibitor ◽  
Target Genes ◽  
Reproductive Function ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Reproductive Process ◽  
Female Mice ◽  
Retinoid Metabolism ◽  
Regulatory Effects

Bikunin, a Kunitz-type protease inhibitor, is found in blood and urine. It has been established by two laboratories independently that the bikunin knockout female mice display a severe reduction in fertility: the cumulus oophorus has a defect in forming the extracellular hyaluronan-rich matrix during expansion. Proteins of the inter-alpha-trypsin inhibitor (ITI) family are eliminated in mice in which the bikunin gene has been inactivated, since bikunin is essential for their biosynthesis. Proteins of the ITI family may contribute to the microenvironment in which ovulation takes place. It is not clear, however, whether a single mechanism affects the reproductive function including ovulation. For identifying the full repertoire of the ITI deficiency-related genes, a cDNA microarray hybridization screening was conducted using mRNA from ovaries of wild-type or bik−/− female mice. A number of genes were identified and their regulation was confirmed by real-time RT-PCR analysis. Our screen identified that 29 (0.7%) and 5 genes (0.1%) of the genes assayed were, respectively, up- and down-regulated twofold or more. The identified genes can be classified into distinct subsets. These include stress-related, apoptosis-related, proteases, signaling molecules, aging-related, cytokines, hyaluronan metabolism and signaling, reactive oxygen species-related, and retinoid metabolism, which have previously been implicated in enhancing follicle development and/or ovulation. Real-time RT-PCR analysis confirmed that these genes were up- and down-regulated two- to tenfold by bikunin knockout. These studies demonstrate that proteins of the ITI family may exert potent regulatory effects on a major physiological reproductive process, ovulation.

Real-time RT-PCR analysis of housekeeping genes in human skeletal muscle following acute exercise

2004 ◽  
Vol 18 (2) ◽  
pp. 226-231 ◽  
Author(s):  
Douglas J. Mahoney ◽  
Kate Carey ◽  
Ming-Hua Fu ◽  
Rodney Snow ◽  
David Cameron-Smith ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Real Time ◽  
Healthy Subjects ◽  
Acute Exercise ◽  
Housekeeping Genes ◽  
Housekeeping Gene ◽  
Cycle Ergometry ◽  
Pcr Analysis ◽  
28S Rrna ◽  
Rt Pcr

Studies examining gene expression with RT-PCR typically normalize their mRNA data to a constitutively expressed housekeeping gene. The validity of a particular housekeeping gene must be determined for each experimental intervention. We examined the expression of various housekeeping genes following an acute bout of endurance (END) or resistance (RES) exercise. Twenty-four healthy subjects performed either a interval-type cycle ergometry workout to exhaustion (∼75 min; END) or 300 single-leg eccentric contractions (RES). Muscle biopsies were taken before exercise and 3 h and 48 h following exercise. Real-time RT-PCR was performed on β-actin, cyclophilin (CYC), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β2-microglobulin (β2M). In a second study, 10 healthy subjects performed 90 min of cycle ergometry at ∼65% of V̇o2 max, and we examined a fifth housekeeping gene, 28S rRNA, and reexamined β2M, from muscle biopsy samples taken immediately postexercise. We showed that CYC increased 48 h following both END and RES exercise (3- and 5-fold, respectively; P < 0.01), and 28S rRNA increased immediately following END exercise (2-fold; P = 0.02). β-Actin trended toward an increase following END exercise (1.85-fold collapsed across time; P = 0.13), and GAPDH trended toward a small yet robust increase at 3 h following RES exercise (1.4-fold; P = 0.067). In contrast, β2M was not altered at any time point postexercise. We conclude that β2M and β-actin are the most stably expressed housekeeping genes in skeletal muscle following RES exercise, whereas β2M and GAPDH are the most stably expressed following END exercise.

scholarly journals Pioglitazone Attenuates Vascular Fibrosis in Spontaneously Hypertensive Rats

PPAR Research ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Dengfeng Gao ◽  
Ning Ning ◽  
Guanghua Hao ◽  
Xiaolin Niu
Keyword(s):  
Real Time ◽  
Immunohistochemical Staining ◽  
Spontaneously Hypertensive Rats ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Hypertensive Rats ◽  
Spontaneously Hypertensive ◽  
Ppar Γ ◽  
Collagen Iii ◽  
Wistar Kyoto

Objective. We sought to investigate whether the peroxisome proliferator-activated receptor-γ (PPAR-γ) ligand pioglitazone can attenuate vascular fibrosis in spontaneously hypertensive rats (SHRs) and explore the possible molecular mechanisms.Methods. SHRs (8-week-old males) were randomly divided into 3 groups (n=8each) for treatment: pioglitazone (10 mg/kg/day), hydralazine (25 mg/kg/day), or saline. Normal male Wistar Kyoto (WKY) rats (n=8) served as normal controls. Twelve weeks later, we evaluated the effect of pioglitazone on vascular fibrosis by Masson’s trichrome and immunohistochemical staining of collagen III and real-time RT-PCR analysis of collagen I, III and fibronectin mRNA.Vascular expression of PPAR-γ and connective tissue growth factor (CTGF) and transforming growth factor-β (TGF-β) expression were evaluated by immunohistochemical staining, western blot analysis, and real-time RT-PCR.Results. Pioglitazone and hydralazine treatment significantly decreased systolic blood pressure in SHRs. Masson’s trichrome staining for collagen III and real-time RT-PCR analysis of collagen I, III and fibronectin mRNA indicated that pioglitazone significantly inhibited extracellular matrix production in the aorta. Compared with Wistar Kyoto rats, SHRs showed significantly increased vascular CTGF expression. Pioglitazone treatment significantly increased PPAR-γ expression and inhibited CTGF expression but had no effect on TGF-β expression.Conclusions. The results indicate that pioglitazone attenuated vascular fibrosis in SHRs by inhibiting CTGF expression in a TGF-β-independent mechanism.

scholarly journals Detection of HER2 Status in Breast Cancer: Comparison of Current Methods with MLPA and Real-time RT-PCR

2013 ◽  
Vol 14 (12) ◽  
pp. 7621-7628 ◽  
Author(s):  
Reza Pazhoomand ◽  
Elahe Keyhan ◽  
Mehdi Banan ◽  
Hossein Najmabad ◽  
Masoud Karimlou ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Real Time ◽  
Her2 Status ◽  
Rt Pcr
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