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Change of the cell cycle after flutamide treatment in prostate cancer cells and its molecular mechanism

Asian Journal of Andrology 2005年第4期7卷 375-380,页

作者： Yong Wang Chen Shao Chang-Hong Shi Lei Zhang Hong-Hong Yue Peng-Fei Wang Bo Yang Yun-Tao Zhang Fan Liu Wei-Jun Qin He Wang Guo-Xing Shao Department of Urology Tangdu Hospital Department of Urology Department of Microbiology Department of Epidemiology Department of Nephrology Xijing Hospital Fourth Military Medical University Xi＇an 710032 Shaanxi China

Aim： To explore the effect of androgen receptor （AR） on the expression of the cell cycle-related genes, such as CDKNIA and BTG1, in prostate cancer cell line LNCaP. Methods： After AR antagonist flutamide treatment and confirmation of its effect by phase contrast microscope and flow cytometry, the differential expression of the cell cycle-related genes was analyzed by a cDNA microarray. The flutamide treated cells were set as the experimental group and the LNCaP cells as the control. We labeled cDNA probes of the experimental group and control group with Cy5 and Cy3 dyes, respectively, through reverse transcription. Then we hybridized the cDNA probes with cDNA microarrays, which contained 8 126 unique human cDNA sequences and the chip was scanned to get the fluorescent values of Cy5 and Cy3 on each spot. After primary analysis, reverse transcription polymerase chain reaction （RT- PCR） tests were carried out to confirm the results of the chips. Results：After AR antagonist flutamide treatment, three hundred and twenty-six genes （3.93 %） expressed differentially, 97 down-regulated and 219 up-regulated. Among them, eight up-regulated genes might be cell cycle-related, namely CDCIO, NRAS, BTG1, Wee1, CLK3, DKFZP564A122, CDKNIA and BTG2. The CDKNIA and BTG1 gene mRNA expression was confirmed to be higher in the experimental group by RT-PCR, while p53 mRNA expression had no significant changes. Conclusion： flutamide treatment might up-regulate CDKN1A and BTG1 expression in prostate cancer cells. The protein expressions of CDKN1A and BTG1 play an important role in inhibiting the proliferation of cancer cells. CDKN1A has a great impact on the cell cycle of prostate cancer cells and may play a role in the cancer cells in a p53-independent pathway. The prostate cancer cells might affect the cell cycle-related genes by activating AR and thus break the cell cycle control.

关键词： prostate cancer LNCaP p21 androgen receptor CDKN1A BTG1 cell cycle genes flutamide

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Induction of liver cytochrome P450 1A2 expression by flutamide in rats

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Acta Pharmacologica Sinica 2005年第11期26卷 1382-1386页

作者： Hai-xue WANG, Xiao LIU, Chang-jiang XU, Xiao-chao MA~3, Jian-er LONG~4, Duan LI~(4,5) ~2 Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 200032, China ~3 State Key Laboratory of Drug Research, Shanghai Institute of Biological Sciences, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 200031, China ~4 Institute of Molecular Virus, School of Basic Medical Science, Fudan University, Shanghai 200031, China Department of Pharmacology School of Pharmacy Fudan University Shanghai China State Key Laboratory of Drug Research Shanghai Institute of Biological Sciences Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai China Institute of Molecular Virus School of Basic Medical Science Fudan University Shanghai China

Aim: To investigate the modulation of liver cytochrome P450 1A2 (CYP1A2) expression by giving flutamide to adult rats. Methods: Rats were given 50, 100, and 200 mg/kg po of flutamide for 2 weeks. Liver CYP 1A2 mRNA was measured using reverse transcription-polymerase chain reaction. CYP1A2 protein was detected using immunoblotting. CYPl A2 activity was assayed using high performance liquid chromatography, with caffeine as the CYPl A2 substrate. Results: CYP1A2 mRNA levels after flutamide treatment at 100 mg/kg and 200 mg/kg were, respectively, 1.86 and 3.11-fold higher than those of the control. Correspondingly, CYP1A2 protein increased 1.78 and 2.89-fold and CYPl A2 activity increased approximately 1.65 and 2.83-fold, respectively, relative to controls. flutamide treatment at 50 mg/kg had no significant effect on CYP1A2 mRNA, protein, or enzyme activity. Conclusion: Giving rats flutamide induced liver CYP1A2 expression in a dose-dependent manner.

关键词： flutamide cytochrome P450 1A2 induction

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flutamide-induced alterations in transcriptional profiling of neonatal porcine ovaries

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Journal of Animal Science and Biotechnology 2019年第3期10卷 618-632页

作者： Katarzyna Knapczyk-Stwora Anna Nynca Renata E.Ciereszko Lukasz Paukszto Jan P.Jastrzebski Elzbieta Czaja Patrycja Witek Marek Koziorowski Maria Slomczynska Department of Endocrinology Institute of Zoology and Biomedical ResearchJagiellonian University in KrakowGronostajowa 9 Street30-387 KrakowPoland Laboratory of Molecular Diagnostics Faculty of Biology and BiotechnologyUniversity of Warmia and Mazury in OlsztynOlsztynPoland Department of Animal Anatomy and Physiology Faculty of Biology and BiotechnologyUniversity of Warmia and Mazury in OlsztynOlsztynPoland Department of Plant Physiology Genetics and BiotechnologyFaculty of Biology and BiotechnologyUniversity of Warmia and Mazury in OlsztynOlsztynPoland Department of Physiology and Reproduction of Animals University of RzeszowRzeszowPoland

Background: Androgens are involved in the regulation of ovarian development during fetal/neonatal life.Environmental chemicals displaying anti-androgenic activities may affect multiple signal transduction pathways by blocking endogenous androgen action.The aim of the current study was to examine effects of the anti-androgen flutamide on the expression of coding transcripts and long non-coding RNAs(lncRNAs) in neonatal porcine ovaries.By employing RNA-Seq technology we aimed to extend our understanding of the role of androgens in neonatal folliculogenesis and examine the impact of the anti-androgen flutamide on ovarian function.Method: Piglets were subcutaneously injected with flutamide(50 mg/kg BW) or corn oil(controls) between postnatal days 1 and 10(n = 3/group).Ovaries were excised from the 11-day-old piglets and total cellular RNAs were isolated and sequenced.Results: flutamide-treated piglet ovaries showed 280 differentially expressed genes(DEGs;P-adjusted flutamide-affected lncRNAs were associated with intracellular transport and cell division.Conclusions: Our results suggest that neonatal exposure of pigs to flutamide alters the expression of genes involved in ovarian cell proliferation,ovarian steroidogenesis and oocyte fertilization,which in turn may affect female reproduction in adult life.

关键词： flutamide LncRNA Ovary Pig RNA-Seq Transcriptome

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Treatment and five-year follow-up of type A insulin resistance syndrome:A case report

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World Journal of Clinical Cases 2022年第8期10卷 2522-2528页

作者： Yong-Hua Chen Qing-Qing Chen Chun-Lin Wang Department of Pediatrics the First Affiliated HospitalZhejiang University School of MedicineHangzhou 310003Zhejiang ProvinceChina

BACKGROUND Type A insulin resistance syndrome(TAIRS)is a rare disorder characterized by severe insulin resistance due to defects in insulin receptor signaling.No specific drugs are available for the treatment of TAIRS.We report a case of TAIRS successfully treated with pioglitazone and flutamide for 5 years.CASE SUMMARY We present the rare case of a female patient aged 11 years and 9 mo with type A insulin resistance and an INSR heterozygous mutation(c.3614 C>T),who was treated with a combination of pioglitazone and flutamide.This treatment regimen reduced hemoglobin A1 c,fasting insulin and androgen levels.CONCLUSION Pioglitazone attenuated insulin resistance in this patient with TAIRS,and flutamide ameliorated masculinization.

关键词： Type A insulin resistance syndrome Treatment Pioglitazone flutamide Case report

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Novel flutamide regulated genes in the rat veritral prostate： differential modulation of their expression by castration and flutamide treatments

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Asian Journal of Andrology 2007年第6期9卷 801-808页

作者： Anil M Limaye Irfan Asangani Namrata Bora Paturu Kondaiah Department of Molecular Reproduction Development and Genetics Indian Institute of Science Bangalore 560012 India

Aim： To identify flutamide regulated genes in the rat ventral prostate. Methods： Total RNA from ventral prostates of control and flutamide treated rats were isolated. Differentially expressed transcripts were identified using differential display reverse transcriptase polymerase chain reaction. The effect of castration on the expression of flutamideregulated transcripts was studied. Results： We have identified β2-microglobulin, cytoplasmic FMR1 interacting protein 2 and pumilio 1 as flutamide induced and spermine binding protein and ribophorin Ⅱ as flutamide repressed targets in the rat ventral prostate. Although flutamide treatment caused an induction of pumilio 1 rnRNA, castration had no effect. Conclusion： Castration and flutamide treatments exert differential effects on gene expression. flutamide might also have direct AR independent effects, which might have implications in the emergence of androgen independent prostate cancer and the failure of flutamide therapy.

关键词： androgens antiandrogen castration differential display reverse transcriptase polymerase chain reaction flutamide prostate

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Frequent Episodic Vertigo is an Unexpected Side Effect of flutamide

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Pharmacology & Pharmacy 2011年第4期2卷 338-340页

作者： Jiann-Jy Chen Dem-Lion Chen 不详

An 82-year-old male suffered from prostatic cancer five years ago. Since then, he has taken flutamide and was bothered with episodic vertigo (EV) every morning. In order to treat prostatic cancer, flutamide was not discontinued, but conservative treatment and life-style change were recommended. Finally, EV actually subsided. Herein, we report the rare case, in which EV was an unexpected side effect of flutamide. Herein we review his whole history, physical examination, vestibular function test, electronystagmogram, caloric test, awake encephalogram, blood examinations, color-coded duplex ultrasonogram and magnetic resonance imaging/angiogram to suggest a mechanism of flutamide responsible for EV.

关键词： flutamide Episodic Vertigo Prostatic Cancer

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Protective mechanism of testosterone on cognitive impairment in a rat model of Alzheimer's disease

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Neural Regeneration Research 2019年第4期14卷 649-657页

作者： Xu-Sheng Yan Zhan-Jun Yang Jian-Xin Jia Wei Song Xin Fang Zhi-Ping Cai Dong-Sheng Huo He Wang Department of Human Anatomy Baotou Medical College School of Health Sciences University of NewcastleNewcastle Australia

Cognitive dysfunction in Alzheimer's disease is strongly associated with a reduction in synaptic plasticity, which may be induced by oxidative stress. Testosterone is beneficial in learning and memory, although the underlying protective mechanism of testosterone on cognitive performance remains unclear. This study explored the protective mechanism of a subcutaneous injection of 0.75 mg testosterone on cognitive dysfunction induced by bilateral injections of amyloid beta 1–42 oligomers into the lateral ventricles of male rats. Morris water maze test results demonstrated that testosterone treatment remarkably reduced escape latency and path length in Alzheimer's disease rat models. During probe trials, testosterone administration significantly elevated the percentage of time spent in the target quadrant and the number of platform crossings. However, flutamide, an androgen receptor antagonist, inhibited the protective effect of testosterone on cognitive performance in Alzheimer's disease rat models. Nissl staining, immunohistochemistry, western blot assay, and enzyme-linked immunosorbent assay results showed that the number of intact hippocampal pyramidal cells, the dendritic spine density in the hippocampal CA1 region, the immune response and expression level of postsynaptic density protein 95 in the hippocampus, and the activities of superoxide dismutase and glutathione peroxidase were increased with testosterone treatment. In contrast, testosterone treatment reduced malondialdehyde levels. flutamide inhibited the effects of testosterone on all of these indicators. Our data showed that the protective effect of testosterone on cognitive dysfunction in Alzheimer's disease is mediated via androgen receptors to scavenge free radicals, thereby enhancing synaptic plasticity.

关键词： nerve regeneration Alzheimer’s disease testosterone cognitive dysfunction synaptic plasticity free radicals Morris water maze androgen receptor flutamide postsynaptic density protein 95 amyloid beta 1–42 neurodegenerative change neural regeneration

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Prior switching to a second-line nonsteroidal antiandrogen does not impact the therapeutic efficacy of abiraterone acetate in patients with metastatic castration-resistant prostate cancer： a real-world retrospective study

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Asian Journal of Andrology 2018年第6期20卷 545-550页

作者： Jin-G Zhao Jian-Dong Liu Peng-Fei Shen Xin Tang Guang-Xi Sun Xing-Ming Zhang Jun-Ru Chen Kun-Peng Shu Ming Shi Hao Zeng Department of Urology Institute of Urology West China Hospital Sichuan University Chengdu 610041 China Department of Oncology West China Hospital Sichuan University Chengdu 610041 China

Even in the era of novel targeted agents, switching to a second-line nonsteroidal antiandrogen （NSAA） is still widely used in treating metastatic castration-resistant prostate cancer （mCRPC）, especially in undeveloped countries. However, whether prior treatment with a second-line NSAA would impact the efficacy of abiraterone acetate （Abi） remains uncertain, in the current study, 87 mCRPC patients treated with Abi were analyzed. Among them, 21 were treated with a second-line NSAA （from bicalutamide to flutamide） before receiving abiraterone, while the remaining 66 received Abi directly. Therapeutic efficacy of Abi was compared between those with and without prior second-line NSAA using Kaplan-Meier curves, log-rank test, and Cox regression models. The therapeutic efficacy of Abi was similar between those with or without the prior switching treatment of flutamide, in terms of either prostate-specific antigen progression-free survival （PSA-PFS, 5.5 vs 5.6 months, P = 0.967）, radiographic progression-free survival （rPFS, 12.8 vs 13.4 months, P = 0.508）, overall survival （OS, not reached vs 30.6 months, P = 0.606）, or PSA-response rate （71.4% [15121] vs 60.6% [40166], P = 0.370）. This is the first time that the impact of prior switching of treatment to a second-line NSAA on the efficacy of Abi in mCRPC patients has been addressed. Our data support that, use of prior sequential bicalutamide and flutamide does not seem to preclude response to abiraterone, although larger cohort studies and, ideally, a randomized controlled trial are needed. These findings will facilitate doctors＇ decision-making in the treatment of mCRPC patients, especially for those with previous experience of switching NSAA second-line treatments in the clinic.

关键词： abiraterone antiandrogen castration-resistant prostate cancer flutamide

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Effect of testosterone on morphine withdrawal syndrome in rats

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Asian Journal of Andrology 2008年第5期10卷 765-769页

作者： Ali Reza Mohajjel Nayebi Hassan Rezazadeh Department of Pharmacology and Toxicology Faculty of Pharmacy Tabriz University of Medical Sciences Tabriz 51664 Iran

Aim： To determine whether testosterone is involved in morphine withdrawal syndrome （WS）. Methods： In order to induce dependency, rats were treated with subcutaneous injection of morphine （days 1-2, 5 mg/kg; days 3-5, 7.5 mg/kg; days 6-8, 10 mg/kg）, and after the last dose of morphine （day 8） WS was induced by intraperitoneal injection of naloxone （1 mg/kg）. Wet dog shake （WDS）, abdomen writhing （AW）, and jumps （J） were recorded as indicators of WS. Results： The severity of WDS, AW, and J in male rats was greater than that in females. Accordingly, in 4-week castrated and flutamide-treated （10 mg/kg/day for 8 days, i.p.） male rats, WDS, AW, and J were significantly decreased compared to male control rats. Testosterone replacement therapy （10 mg/kg/day for 8 days, i.m.） in 4-week castrated rats restored the severity of WDS, AW, and J behaviors to the level of non-castrated male rats, whereas testosterone potentiated the WDS behavior in non-castrated male rats. Conclusion： It can be concluded that testosterone might be effectively involved in morphine WS.

关键词： testosterone castration flutamide morphine withdrawal syndrome

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flutamide对前列腺癌细胞分化抑制因子1表达影响

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青岛大学医学院学报 2015年第1期51卷 94-96页

作者：王德举郭晓慧于小玲青岛大学医学院泌尿外科山东青岛266021 青岛市城阳区第二人民医院青岛市城阳区妇幼保健所青岛大学医学院病理生理学教研室

目的了解雄激素受体阻断剂flutamide对前列腺癌细胞分化抑制因子1（ID1）表达的影响及意义。方法在培养的前列腺癌LNCaP和PC3细胞中分别加入flutamide,用实时荧光定量反转录聚合酶链式反应（RTPCR）方法检测ID1mRNA相对定量表达;用Weste... 详细信息

目的了解雄激素受体阻断剂flutamide对前列腺癌细胞分化抑制因子1（ID1）表达的影响及意义。方法在培养的前列腺癌LNCaP和PC3细胞中分别加入flutamide,用实时荧光定量反转录聚合酶链式反应（RTPCR）方法检测ID1mRNA相对定量表达;用Western Blotting方法检测ID1蛋白表达,以不加flutamide细胞作为对照组。结果与对照组比较,flutamide组LNCaP和PC3细胞ID1 mRNA相对定量明显减少,LNCaP细胞中ID1蛋白表达明显减少,差异有显著性（t=3.09~34.91,P〈0.05）。flutamide组LNCaP中ID1 mRNA相对定量、ID1蛋白表达低于PC3细胞,差异有显著性（t=2.57、12.52,P〈0.05）。结论 flutamide能抑制前列腺癌细胞中ID1的表达,ID1的表达可能受雄激素-受体途径的调控。

关键词：前列腺肿瘤 LNCaP细胞 PC3细胞细胞分化抑制因子1 flutamide

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