Quantification of androgen receptor mRNA in tissues by competitive co-amplification of a template in reverse transcription—Polymerase chain reaction

Malucelli, Alberto; Sauerwein, H.; Pfaffl, Michael W.; Meyer, Heinrich

doi:10.1016/0960-0760(96)00077-5

Cited by 9 publications

(10 citation statements)

References 26 publications

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“…The effects of steroid hormones are mediated through interaction with specific intracellular receptors, which are members of the nuclear receptor family [4,5]. Numerous tissues were shown to express mRNA transcripts for both estrogen receptors, subtypes estrogen receptor ␣ (ER␣) and ␤ (ER␤) [6][7][8][9], androgen receptor (AR) [10][11][12] and progestin receptor (PR) [3,13]. However simultaneous occurrence of noma xenograft (CWR22R) that was serially propagated in mice after castration-induced regression and relapse of the parental, androgen-dependent CWR22 xenograft [22].…”

Section: Introductionmentioning

confidence: 99%

Characterisation of steroid receptor expression in the human prostate carcinoma cell line 22RV1 and quantification of androgen effects on mRNA regulation of prostate-specific genes

Hartel

Didier

Ulbrich

et al. 2004

The Journal of Steroid Biochemistry and Molecular Biology

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Section: Introductionmentioning

confidence: 99%

Characterisation of steroid receptor expression in the human prostate carcinoma cell line 22RV1 and quantification of androgen effects on mRNA regulation of prostate-specific genes

Hartel

Didier

Ulbrich

et al. 2004

The Journal of Steroid Biochemistry and Molecular Biology

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“…10 Androgen and ␤-actin (control) primers were designed from the published sequence of the human genes 11,12 : AR (sense), 5Ј-AGATGGGCTTGACTTTCCCAGAAAG-3Ј; AR (antisense), 5Ј-ATGGCTGTCATTCAGTACTCCTGGA-3Ј.…”

Section: Rna Extraction and Measurement Of Ar Mrna Levelsmentioning

confidence: 99%

Androgen Receptor Expression Is Greater in Macrophages From Male Than From Female Donors

et al. 2000

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Background-Male sex is an independent risk factor for the extent and severity of atherosclerosis. The influence of androgens on foam cell formation, a key event in atherogenesis, has not yet been investigated. Methods and Results-Primary human monocytes were allowed to differentiate into macrophages. RNA was then extracted from healthy male-donor (nϭ8) and premenopausal female-donor (nϭ8) macrophages, and message for the androgen receptor (AR) was examined by RT-PCR. There was a significantly higher level of AR mRNA in macrophages isolated from men than in those from women (0.64Ϯ0.06 versus 0.15Ϯ0.02 amol/g total RNA; PϽ0.001). AR mRNA levels were similar in macrophages from postmenopausal and premenopausal women (Pϭ0.16). The functional consequence of this sex difference was then explored. Lipid-loading studies were performed on male (nϭ9) macrophages treated with the androgen dihydrotestosterone (DHT) and/or the AR antagonist hydroxyflutamide. These showed that DHT caused a dose-dependent and receptor-mediated increase in macrophage cholesteryl ester content (109Ϯ10%, 117Ϯ3%, and 120Ϯ4% for 4, 40, and 400 nmol/L DHT, respectively, as a percentage of control, Pϭ0.002; 95Ϯ8% for DHT with hydroxyflutamide, Pϭ0.58 versus controls). By contrast, there was no significant effect of androgen on lipid loading in female-donor macrophages (PϾ0.2 versus controls). Conclusions-Sex differences in androgen-mediated macrophage lipid loading may contribute to the greater prevalence and severity of atherosclerosis in men. en have an earlier onset and higher incidence of atherosclerosis than women. 1 The potential protective effects of estrogens on vascular structure and function have been studied extensively. Much less work, however, has addressed the possible effects of androgens on atherogenesis. In animal models, androgen treatment may increase plaque formation in the aorta and coronary vessels. 2,3 Furthermore, in humans, we have recently demonstrated that androgens enhance monocyte adhesion to endothelial cells 4 and are associated with impaired vascular reactivity. 5 Foam cell formation is a key early event in atherosclerosis and is largely due to the uptake of modified lipoproteins, principally LDL, by peripheral blood monocyte-derived macrophages (MDMs) in the arterial wall. 6,7 The possibility that androgen receptors (ARs) and androgen-related intracellular pathways may be involved in the process of lipid loading and foam cell formation in atherosclerosis has not been explored previously. We therefore investigated whether the AR is expressed in primary human MDMs and whether there is a sex difference in its expression and have examined the biological effects of androgen stimulation on cholesteryl ester (CE) formation/lipid loading in these cells. Methods Isolation of Human MonocytesWhite cell concentrates (Red Cross Blood Bank) were obtained from the peripheral blood of individual healthy men (20 to 58 years old) and premenopausal (18 to 45 years old) and postmenopausal (45, 55, and 72 years old) women. The postmenopau...

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“…The primer sequences are summarized in Table 1. AR (Malucelli et al., 1996), PR, ERα (Pfaffl et al., 2001) and ERβ were designed in the region of the receptor ligand binding domain to produce 172, 227, 234 and 242 bp amplification products, respectively. IGF‐1 primers were designed to produce a 240 bp amplification product in the highly conserved region of exons 3 and 4 (Gilmour et al., 1992) coding for the mature IGF‐1 protein (Pfaffl et al., 1998).…”

Section: Methodsmentioning

confidence: 99%

Effects of Synthetic Progestagens on the mRNA Expression of Androgen Receptor, Progesterone Receptor, Oestrogen Receptor α and β, Insulin‐like Growth Factor‐1 (IGF‐1) and IGF‐1 Receptor in Heifer Tissues

Pfaffl

Daxenberger

Hageleit

et al. 2002

Journal of Veterinary Medicine Series A

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Synthetic progestagens like melengestrol acetate (MGA) are widely used for oestrus synchronization and for growth promotion in cattle production. The metabolic effects exceed its primary potency as a progestagen. It is speculated that MGA stimulates follicle development and thereby endogenous oestrogen production, but inhibits ovulation. To investigate the dose-dependent effects on mRNA expression levels, six heifers were fed for 8 weeks with different levels of MGA (0.5, 1.5, 5 mg) daily and two heifers served as controls. The expression of steroid receptor mRNA [androgen receptor (AR), progesterone receptor (PR), oestrogen receptor (ER) ERalpha and ERbeta], insulin-like growth factor-1 (IGF-1) and its receptor were quantified in liver, neck (m. splenius) and shoulder muscularity (m. deltoideus). Plasma concentrations of IGF-1 were quantified by radioimmunoassay. In treated animals the MGA plasma levels were elevated over the complete treatment period, corresponding to the MGA treatment concentrations. IGF-1 concentrations of control animals were at constant levels. Plasma levels for oestradiol (E2) and IGF-1 were increased in the low MGA treatment group. Overdosed MGA decreased progesterone (P4) and E2 levels. To quantify the IGF-1 and all receptor mRNA transcripts, sensitive and reliable real-time reverse transcription-polymerase chain reaction (RT-PCR) quantification methods were developed and validated in the LightCycler. A dose-dependent relationship between increasing MGA concentration and mRNA expression was observed in liver for AR and IGF-1 receptor, and in neck muscularity for IGF-1. ERalpha in liver and neck muscle showed a trend of increasing expression.

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Quantification of androgen receptor mRNA in tissues by competitive co-amplification of a template in reverse transcription—Polymerase chain reaction

Cited by 9 publications

References 26 publications

Characterisation of steroid receptor expression in the human prostate carcinoma cell line 22RV1 and quantification of androgen effects on mRNA regulation of prostate-specific genes

Characterisation of steroid receptor expression in the human prostate carcinoma cell line 22RV1 and quantification of androgen effects on mRNA regulation of prostate-specific genes

Androgen Receptor Expression Is Greater in Macrophages From Male Than From Female Donors

Effects of Synthetic Progestagens on the mRNA Expression of Androgen Receptor, Progesterone Receptor, Oestrogen Receptor α and β, Insulin‐like Growth Factor‐1 (IGF‐1) and IGF‐1 Receptor in Heifer Tissues

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