Flutamide对前列腺癌细胞分化抑制因子1表达影响

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。

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

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.

Flutamide-induced alterations in transcriptional profiling of neonatal porcine ovaries

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 < 0.05 and log2 fold change ≥1.0) and 98 differentially expressed lncRNAs(DELs;P-adjusted < 0.05 and log2 FC ≥ 1.0).The DEGs were assigned to GO term,covering biological processes,molecular functions and cellular components,which linked the DEGs to functions associated with cellular transport,cell divisions and cytoskeleton.In addition,STRING software demonstrated strongest interactions between genes related to cell proliferation.Correlations between DEGs and DELs were also found,revealing that a majority of the genes targeted by the 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.

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

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.

Frequent Episodic Vertigo is an Unexpected Side Effect of Flutamide

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.

缓退瘤治疗前列腺癌

目的：探讨缓退瘤（Ｆｌｕｔａｍｉｄｅ）与手术或药物去势联合治疗前列腺癌的疗效。方法：回顾性分析１８例手术或药物去势联合缓退瘤治疗的前列腺癌患者的临床资料。结果：病理确诊后１４例行手术去势及缓退瘤治疗，４例行药物去势加缓退瘤治疗，１６例获得显著效果，１例出现肝功能损害，１例出现缓退瘤撤除综合征。结论：联合治疗可最大限度地阻断雄激素对前列腺的作用，使前列腺癌细胞迅速凋亡，从而延长患者的生存期。

血清前列腺特异性抗原在雄激素阻断治疗前列腺癌中的意义

目的探讨血清前列腺特异性抗原(PSA)在雄激素阴断治疗前列腺癌中的作用与意义。方法根据检查结果按Whitmore-Jewett标准进行临床分期:B期3例,C期14例,D期7例,双侧睾丸切除术后 1周开始服用Flutamide,剂量为250 mg,每8 h1次。每月复查血清PSA及前列腺体积。结果随访3个月～3年。 1个月后PSA水平明显下降:6个月后B超显示前列腺体积明显缩小;2例肺部转移,5个月后复查发现肺部肿块缩小;5例骨转移伴骨痛,1个月后骨痛明显缓解,6个月后ECT及CT显示骨转移灶缩小。结论去势加 Flutamide是一种治疗中晚期前列腺癌安全、有效的方法,可根据PSA的水平来调节用药及预后监测。

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

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.

雄激素受体抑制剂通过AR信号通路对胶质瘤生物学特性的影响

目的研究雄激素受体(androgen receptor,AR)抑制剂(flutamide)对人胶质瘤细胞增殖、凋亡、侵袭的影响。方法采用不同浓度的flutamide(低浓度1.0×10^(-8)、中浓度1.0×10^(-7)、高浓度1.0×10^(-6)mol/L)作用于胶质瘤细胞系U251,Real Time-PCR和Western Blot检测AR m RNA及AR蛋白的表达情况;Cell Counting Kit-8法测定flutamide对胶质瘤细胞增殖的影响;Annexin V-7AAD/PE双染检测flutamide诱导胶质瘤细胞凋亡的作用;Transwell测定flutamide对胶质瘤细胞侵袭能力的改变。结果在体外胶质瘤细胞的增殖、侵袭能力能被AR抑制剂(flutamide)显著抑制且凋亡增加,与药物浓度正相关。结论 AR抑制剂(flutamide)能明显抑制胶质瘤细胞的增殖与侵袭,并且诱导细胞凋亡,AR可能是引起胶质瘤发生、发展的危险因素。

Effect of testosterone on morphine withdrawal syndrome in rats

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.

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

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.

Sexual Function in Men with Castrate Levels of Testosterone: Observations of a Subgroup of Sexually Active Men with Prostate Cancer Undergoing Androgen Deprivation Therapy

Purpose: To identify possible factors that influence sexual function in men undergoing maximal androgen deprivation therapy (ADT). Patients and Methods: A descriptive exploration was performed looking at characteristics of twenty-two men reporting sexual activity after nine months of maximal ADT. This previously published Phase II study, involved 250 prostate cancer patients undergoing intermittent ADT. An analysis between this cohort and the group that did not maintain sexual function was performed to ascertain if age, testosterone level, functional status or maintenance of quadriceps strength had an impact upon sexual function. Results: There was no difference in age, testosterone level or ECOG performance status between the sexually active and non-sexually active groups. Over the course of 9 months of ADT, the sexually active group appeared to maintain quadriceps muscle strength as measured with physical stands, and maintained overall health as measured by quality of life questionaries, compared to the non-sexually active group. Conclusions: This retrospective study suggests that exercise during ADT may reduce the impact of ADT on sexual function. This warrants further testing, and could be the focus of future randomised controlled trials.

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

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.