OBJECTIVETo determine the effects of Chinese herbal monomers such as baicalin, berberine, and matrine on the androgen receptor (AR) mRNA expression in SZ95 sebocytes in vitro and to explore the possible mechanism of using traditional Chinese medicines to treat acne.

METHODSSZ95 sebocytes were cultured and then treated with berberine, baicalin, matrine, and 13-cis-retinoic acid for 24 hours. Reverse transcription polymerase chain reaction was applied to detect the changes of AR.

RESULTAR mRNA was downregulated by 13-cis-retinoic acid of 1 x 10(-5) mol/L and 1 x 10(-6) mol/L, and by baicalin of 1 x 10(-4) mol/L (P < 0.05).

CONCLUSION13-cis-retinoic acid and baicalin may exert antiandrogenitic action by inhibiting AR mRNA expression in human sebocytes.

Androgen Antagonists ; pharmacology ; Cell Line ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; pharmacology ; Humans ; RNA, Messenger ; biosynthesis ; Receptors, Androgen ; biosynthesis ; genetics ; Skin ; cytology

OBJECTIVESTo study the distribution and potential function of androgen receptor (AR) mRNA and AR in rat submaxillary.

METHODSIn situ hybridization using digoxigenin-labled oligonucleotide probes, cell culture and radio-immunoassay were performed to localize the AR and detect the concentration of epidermal growth factor (EGF) in culturing supernant.

RESULTSAR mRNA hybridization signals were detected in glandular epithelial cells of serous acinus and epithelial cells in all gland ducts. The signals distributed in cytoplasma of all positive cells with negative nuclei; Administration of testosterone can significantly increase the level of EGF (P < 0.05).

CONCLUSIONSThe rat submaxillary not only is a target organ of androgen but also can product AR by itself. When androgen combined with androgen receptor and can submaxillary function can be infected and can result in the elevation of the level of EGF secreted.

Animals ; Epidermal Growth Factor ; biosynthesis ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; genetics ; physiology ; Submandibular Gland ; metabolism

AIMTo examine the physiological role of the androgen receptor (AR) in the PC-3 cell line by transfecting full-length functional AR cDNA driven by its natural human AR promoter.

METHODSWe generated an AR-expressing PC-3(AR)9 stable clone that expresses AR under the control of the natural human AR promoter and compared its proliferation to that of the PC-3(AR)2 (stable clone that expresses AR under the control of the cytomegalovirus (CMV) promoter, established by Heisler et al.) after androgen treatment.

RESULTSWe found that dihydrotestosterone (DHT) from 0.001 nmol/L to 10 nmol/L induces cell cycle arrest or inhibits proliferation of PC-3(AR)2 compared with its vector control, PC-3(pIRES). In contrast, PC-3(AR)9 cell growth slightly increased or did not change when treated with physiological concentrations of 1 nmol/L DHT.

CONCLUSIONThese data suggest that intracellular control of AR expression levels through the natural AR promoter might be needed for determining AR function in androgen-independent prostate cancer (AIPC) PC-3 cells. Unlike previous publications that showed DHT mediated suppression of PC-3 growth after transfection of viral promoter-driven AR overexpression, we report here that DHT-mediated PC-3 proliferation is slightly induced or does not change compared with its baseline after reintroducing AR expression driven by its own natural promoter, as shown in PC-3(AR)9 prostate cancer cells.

Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; DNA, Complementary ; biosynthesis ; Dihydrotestosterone ; pharmacology ; Humans ; Male ; Promoter Regions, Genetic ; Prostatic Neoplasms ; metabolism ; pathology ; Receptors, Androgen ; biosynthesis ; genetics ; Transfection

When we try to establish the gene recombinant yeast cell to screen the androgenic endocrine disruptors, the key procedure is the androgen receptor (AR) expression in the yeast cell. For this purpose, we obtained the GPD (glyceraldehyde-3-phosphote dehydrogenase) promoter from the yeast genosome of W303-1A using PCR system and inserting it into Swa I and BamH I sites of pYestrp2. The new constructed vector was named pGPD. The V5 epitope tag DNA with a 5'-BamH I and a 3'-EcoR I sticky end was cloned into the corresponding site of the pGPD vector to yield the vector of pGPDV5. The 2 723 bp full length AR ORF amplified by PCR from pcDNA3.1/AR was fused to V5 epitope tag DNA in pGPDV5 to give the AR yeast expression vector of pGPDV5/AR. This fused vector was transformed into the yeast cell (W303-1A). Western blot was used to detect the V5 fused protein of AR, in the protocol of which the primary monoclonal antibody (IgG(2a)) of mouse anti-V5 and the polyclonal secondary antibody of goat anti-mouse (IgG) linked to horseradish peroxidase (HRP) were used to detect the specific protein in the given sample of the transformed yeast extract. The result showed that the fused protein of AR was expressed successfully in the yeast cell.
Base Sequence ; Endocrine Disruptors ; analysis ; Epitopes ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases ; genetics ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Receptors, Androgen ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Yeasts ; genetics ; metabolism

OBJECTIVETo investigate if there is abnormal expression of androgen receptor (AR) in adult diabetic rats.

METHODSThe diabetic model rats were induced by using streptozotocin (STZ), which were divided into three groups: control (group C), diabetes (group D) and diabetes with insulin replacement group (group ID). The mRNA and protein expressions of androgen receptor in testis, epididymis and prostate were detected by Northern blot and radioimmunoassay, respectively.

RESULTSSerum testosterone levels and AR mRNA in epididymis of group D or Group ID were lower than those of group C (P <0.05) , respectively, and the protein expression of AR in testis and prostate of group D was lower than that of group C (P <0.05).

CONCLUSIONThe expression of androgen receptor decreased in testis, epididymis and prostate of diabetic rats, which weakened the biological effects of AR, and it might be one of the causes that resulted in the sexual and productive dysfunction in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; Epididymis ; metabolism ; Male ; Prostate ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Androgen ; biosynthesis ; genetics ; Testis ; metabolism

OBJECTIVETo investigate the effects of perinatal thyroid hormone deficiency on the expression of androgen receptor (AR) mRNA in cerebral cortex and hippocampus of rats.

METHODSPerinatal hypothyroidism was induced by the administration of propylthiouracil (PTU) solution to the dams by gavage (50 mg/d) beginning at embryonic d15 throughout the lactational period. In the T(4) injected group hypothyroid rats were injected intraperitoneally with levothroxine (L-T(4)) 2 microg/100 g BW daily, starting from the day of birth. Cerebral cortex and hippocampus specimen were collected from controls,hypothyroid and T(4)-injected hypothyroid rats on postnatal d1, 5, 10, 15 and 20. Quantification of ARmRNA in cerebral cortex and hippocampus was performed with competitive RT-PCR using internal and external standardization.

RESULTAge-related increasing ARmRNA levels were observed in neonatal rats, and those in male animals were significantly higher. AR expression was higher in the hippocampus than in the cerebral cortex. ARmRNA levels in the hypothyroid pups were lower than those in age-matched controls. The mRNA levels in the T(4)-injected hypothyroid pups were significantly higher compared with the age-matched hypothyroid pups, but in hippocampus ARmRNA expression did not reach normal levels in male rats at d10 and d20, in female at d15 and d20.

CONCLUSIONThe expression of ARmRNA decreases in brain of rats with perinatal hypothyroidism. Treatment with thyroid hormone can recover ARmRNA expression in cerebral cortex, but not in hippocampus.

Animals ; Animals, Newborn ; Cerebral Cortex ; metabolism ; Female ; Hippocampus ; metabolism ; Hypothyroidism ; metabolism ; Pregnancy ; Pregnancy Complications ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Receptors, Androgen ; biosynthesis ; genetics

AIMTo evaluate androgen receptor (AR) expression in clinically localized prostate cancer (PCa).

METHODSSpecimens were studied from 232 patients who underwent radical prostatectomy for clinically localized prostatic adenocarcinoma without neoadjuvant hormonal therapy or chemotherapy at our institution between November 2001 and June 2005. Immunohistochemical study was performed using an anti-human AR monoclonal antibody AR441. The mean AR density in the hot spots of different histological areas within the same sections were compared and the correlation of malignant epithelial AR density with clinicopathological parameters such as Gleason score, tumor, nodes and metastases (TNM) stage and pre-treatment prostate-specific antigen (PSA) value was assessed.

RESULTSAR immunoreactivity was almost exclusively nuclear and was observed in tumor cells, non-neoplastic glandular epithelial cells and a proportion of peritumoral and interglandular stromal cells. Mean percentage of AR-positive epithelial cells was significantly higher in cancer tissues than that in normal prostate tissues (mean +/- SD, 90.0% +/- 9.3% vs. 85.3 +/- ?9.7%, P < 0.001). The histological score yielded similar results. The percentage of AR immunoreactive prostatic cancer nuclei and histological score were not correlated with existing parameters such as Gleason score, tumor, nodes and metastases stage and pre-treatment PSA value in this surgically treated cohort.

CONCLUSIONThe results of the present study suggest that there may be limited clinical use for determining AR expression (if evaluated in hot spots) in men with localized PCa.

Adenocarcinoma ; genetics ; pathology ; surgery ; Aged ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Paraffin Embedding ; Prostate-Specific Antigen ; analysis ; metabolism ; Prostatectomy ; Prostatic Neoplasms ; genetics ; pathology ; surgery ; Receptors, Androgen ; biosynthesis ; genetics

Thirty-two cases of surgically removed astrocytoma were evaluated for the expression of androgen receptors(ARs) immunohistochemically and the relationships between androgen receptors, DNA ploidy pattern, and survival of patients were studied. The cases included 18 grade I/II astrocytomas, 4 anaplastic astrocytomas, and 10 glioblastoma multiforme(GBM). Positive AR was present in 12 out of 32 cases(38%), which consisted of 5 cases in grade I/II(28%), 3 cases in anaplastic astrocytoma(75%), and 4 cases in GBM(40%). For both low and high grade astrocytomas, sex and ploidy pattern were not correlated with expression of the androgen receptors. Androgen receptor expression did not significantly affect the survival time. This study confirms previous reports of a low incidence of androgen receptors in astrocytomas. In addition, it shows that expression of androgen receptors is not correlated with DNA ploidy pattern and survival of patients in astrocytoma.
Adolescent ; Adult ; Aged ; Astrocytoma/*metabolism/pathology ; Brain Neoplasms/*metabolism/pathology ; Child ; Female ; Flow Cytometry ; Glioma/metabolism/pathology ; Human ; Male ; Middle Age ; Ploidies ; Receptors, Androgen/*biosynthesis/genetics ; Support, Non-U.S. Gov't ; Survival Analysis

Thirty-two cases of surgically removed astrocytoma were evaluated for the expression of androgen receptors(ARs) immunohistochemically and the relationships between androgen receptors, DNA ploidy pattern, and survival of patients were studied. The cases included 18 grade I/II astrocytomas, 4 anaplastic astrocytomas, and 10 glioblastoma multiforme(GBM). Positive AR was present in 12 out of 32 cases(38%), which consisted of 5 cases in grade I/II(28%), 3 cases in anaplastic astrocytoma(75%), and 4 cases in GBM(40%). For both low and high grade astrocytomas, sex and ploidy pattern were not correlated with expression of the androgen receptors. Androgen receptor expression did not significantly affect the survival time. This study confirms previous reports of a low incidence of androgen receptors in astrocytomas. In addition, it shows that expression of androgen receptors is not correlated with DNA ploidy pattern and survival of patients in astrocytoma.
Adolescent ; Adult ; Aged ; Astrocytoma/*metabolism/pathology ; Brain Neoplasms/*metabolism/pathology ; Child ; Female ; Flow Cytometry ; Glioma/metabolism/pathology ; Human ; Male ; Middle Age ; Ploidies ; Receptors, Androgen/*biosynthesis/genetics ; Support, Non-U.S. Gov't ; Survival Analysis

OBJECTIVETo investigate the effect of Zilongjin (ZLJ) on human androgen-dependent type of prostate cancer cell line LNCaP.

METHODSMTT assay, flow cytometry and fluorescence microscopy were used to observe the effect of ZLJ in anti-proliferation, cell cycle arresting and apoptosis induction. RT-PCR was used to examine the effect of ZLJ on expressions of prostate marker gene (PSA), androgen receptor (AR), apoptosis related genes (bcl-2 and bax), and Western blot assay was used to detect the effect on protein expression of bcl-2 and bax.

RESULTSZLJ could cause apparent inhibition on proliferation, induce G0/G1 phase arresting and apoptosis in time- and dose-dependent manner on LNCaP cells. The concentration for inhibiting cell growth by 50% (IC50) in 72 hrs was 0.79 mg/ml. ZLJ could down-regulate the expression of PSA, AR, bcl-2 genes and lower bcl-2 protein expression, but showed ineffective on bax protein expression.

CONCLUSIONZLJ displays its anti-tumor effects by way of inhibiting the cell proliferation, arresting the G0/G1 phase, inducing apoptosis, down-regulating PSA, AR, bcl-2 gene expression and lowering bcl-2 protein expressions.

Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Neoplasms, Hormone-Dependent ; metabolism ; pathology ; Prostate-Specific Antigen ; biosynthesis ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Receptors, Androgen ; biosynthesis ; genetics