依立雄胺对激素非依赖型人前列腺癌细胞的体外抑制作用

目的 :观察依立雄胺在体外对激素非依赖型人前列腺癌细胞 (PC3)正常生长及Bcl 2蛋白表达的作用。方法 :用 5 ,15和 4 5 μmol·L- 1的依立雄胺作用PC3细胞 3d或 7d后 ,相差显微镜进行细胞形态学观察 ;苔盼蓝染色活细胞计数 ,绘制 7d的生长曲线 ;应用流式细胞仪分析依立雄胺对PC3细胞凋亡的影响 ;应用免疫组化比较经依立雄胺作用后PC3细胞Bcl 2的表达强度。结果 :依立雄胺在5 ,15和 4 5 μmol·L- 1作用PC3细胞 3d后 ,可使其有明显的皱缩脱壁 ,部分细胞膜破裂 ,并抑制PC3细胞的生长 ,抑制率依次为 19% ,2 6 %和 6 0 % ;流式细胞仪分析显示上述浓度的依立雄胺不诱导细胞凋亡 ,但可抑制细胞于DNA合成 (S)期 ,其百分率分别为 18% ,16 %和 11% ,而对照组为 33% ;免疫组化分析表明依立雄胺也不能降低PC3细胞中Bcl 2的表达。结论 :5 ,15和 4 5 μmol·L- 1的依立雄胺在体外可抑制PC3细胞的生长 ,机制可能为抑制细胞DNA的合成。     

Endothelin-1 as a target for therapeutic intervention in prostate cancer

The endothelins, a family of potentvasoconstricting peptides, have beenimplicated in the pathophysiology ofadvanced prostate cancer. Two endothelinreceptors, ET-A and ET-B are found innormal prostate tissue. Malignant prostatecells are notable for the loss of ET-Breceptors and increased levels ofendothelin-1 [ET-1]; this distortion of theendothelin system may be a significantfactor in the progression of prostatecancer. Proposed roles for endothelin inprostate cancer include growth promotion,apoptosis inhibition, bone formation, andstimulation of nociceptive receptors. ET-1can act alone as a mitogen, but its effectsare greatest as a comitogen with a varietyof growth factors, including basicfibroblast growth factor, insulin-likegrowth factors, and platelet derived growthfactor. Although their exact functions areunclear, ET-1, in conjunction with vascularendothelial growth factor, appears to playa major role in tumor angiogenesis. By avariety of methods, ET-1 alters the balanceof osteoblast and osteoclasts to the favornew bone formation that is characteristicof metastatic disease. Several studiesindicate that the refractory pain ofmetastatic cancer is related to the directnociceptive effects ET-1. These findingssuggest that ET receptors are promisingtherapeutic targets for pharmacologicintervention. Early clinical trialsindicate that the ET-A receptor antagonistused in prostate cancer is reasonably welltolerated with mild but pervasive symptomsrelated to ET-1's vasoconstrictive effects. Results of ongoing clinical trials areeagerly awaited in order to see if thehypothetical promise of ET antagonism willresult in clinical success.     

Atrasentan: targeting the endothelin axis in prostate cancer

Endothelin axis deregulation triggers a series of events that lead to a profound deregulation in cancer cells, including key tumorigenic cellular events such as proliferation, invasion, escape from programmed cell death, new vessel formation, abnormal osteogenesis and the alteration of nociceptive stimuli. Atrasentan is a novel agent that effectively targets this pathway and is able to inhibit and/or reverse several of those events. Biological and clinical activity in patients with prostate cancer has been demonstrated in a Phase III clinical setting by the suppression of markers of biochemical and clinical prostate cancer progression, and by a delay in time to disease progression, especially in patients with bone disease.     

Overexpression of coxsackie and adenovirus receptor inhibit growth of human bladder cancer cell in vitro and in vivo

Aim： To study the effect of the overexpression of coxsackie and the adenovirus receptor （CAR） on the growth of the human bladder cancer cell in vitro and in vivo. Methods： A retroviral vector pLXSN-CAR expressing CAR was constructed and confirmed by restriction enzyme mapping. The pLXSN-CAR vector and con- trol vector pLXSN were transfected into the PT67 packaging cell line to generate retrovirus with high titer. The CAR-negative T24 cell was infected with the pLXSNCAR and the pLXSN retrovirus, respectively. The positive clone cells were selected with G418 for 2 weeks. The expression level of the CAR protein was detected by Western blot assay. T24 cell growth in vitro was determined by 3-（4, 5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay. Anchorage-independent growth was measured by soft-agar colony formation assay. In vivo cell growth was determined by a nude mice xenograft model. Results： The pLXSN-CAR vector containing full-length CAR cDNA was successfully constructed. Western blot analysis showed that a 46 kDa specific band was found in pLXSN-CA-transfected T24 cells. MTT assay identified the growth inhibition of T24/pLXSN-CAR cells. The cell colony forming ability of T24/pLXSNCAR cells was significantly lower than that of T24/pLXSN and parental T24 cells. There was a reduction in the tumor size in the T24/pLXSN-CAR group as compared with that of the T24/pLXSN group and parental T24 group. Conclusion： The overexpression of CAR in T24 bladder cancer cells can inhibit cell growth both in vitro and in vivo.     

Novel triazole derivatives with improved receptor activity and bioavailability properties as ghrelin antagonists of growth hormone secretagogue receptors (WO2012035124): a patent evaluation

The patent claims triazole compounds as inhibitors of the ghrelin receptor, the growth-hormone secretagogue receptor (GHS-R1a). These compounds are potent GHS-R1a ligands with an improved in vitro antagonistic activity against ghrelin receptors of at least factor 3 compared with a representative compound disclosed in a previous patent, and are expected to possess, on the basis of the results of in vitro assays, improved safety and human oral bioavailability with respect to the lead.     

雄激素受体抑制剂的研究进展

雄激素受体与前列腺癌的发生、发展密切相关,是配体依赖的转录因子,其与雄激素结合后发生构象转化形成二聚体而活化,并进入细胞核中与相应的 DNA 反应元件结合,促进靶基因的转录,进而促进细胞的增殖。在前列腺癌组织中,雄激素受体的增殖或突变能使其对血清中较低含量的雄激素敏感,是前列腺癌恶化的主要原因。设计合成对野生型及突变型雄激素受体具有较好抑制活性的药物是治疗前列腺癌的首要策略。该文综述了雄激素受体的结构、功能及其抑制剂的研究进展,期望为新型抗前列腺癌药物的设计研究提供参考。     

ZD4054: a specific endothelin A receptor antagonist with promising activity in metastatic castration-resistant prostate cancer

Background: Overexpression of the endothelin A (ET_A) receptor has been found in a number of human cancer cell lines. Activation of the ET_A receptor by endothelin-1 (ET-1) promotes cell proliferation and survival in these tumors, whereas activation of the endothelin B (ET_B) receptor results in an opposing effect. Therefore, blockade of ET_A may have antitumor effects, while sparing ET_B-mediated effects such as induction of apoptosis and clearance of ET-1. Objective: ZD4054 is an orally bioavailable, specific ET_A antagonist currently being investigated in prostate cancer. In receptor-binding studies, ZD4054 only bound to ET_A, with no binding detected towards ET_B. Prostate cancer cell lines are known to produce ET-1 and there is a relative increase in expression of ET_A to ET_B in these cancers. There is also an association of greater ET_A expression in higher grade versus lower grade tumors, suggesting that ET_A may be involved in the malignant transformation process. As ET-1 may also mediate nociceptive effects and osteoblastic effects, there is much interest in clinically assessing ZD4054 in prostate cancer. Methods: The data describing the endothelin axis, the role of ET_A and ET_B in malignancy, and the effects of ET_A antagonist ZD4054 in prostate cancer, as demonstrated in preclinical and clinical studies, are reviewed. Results: Further investigation of ZD4054 in prostate cancer is warranted, and Phase III trials are already planned in patients with non-metastatic castrate-resistant prostate cancer (CRPC) with rising prostate specific antigen values, metastatic (asymptomatic) CRPC, and in metastatic CRPC in combination with docetaxel, assessing either differences in progression-free survival and overall survival or overall survival alone.     

The prospects of antagonizing the growth hormone secretagogue receptor to treat obesity

Background: Ghrelin is an endogenous ligand of the growth hormone secretagogue receptor (GHS-R) that functions as a short-term meal initiator and a long-term energy balance regulator. Antagonizing GHS-R could be a method to treat obesity. Objective: To review the published in vivo characterization of GHS-R antagonists between 2005 and 2008 and evaluate the validity of antagonizing GHS-R as a therapeutic strategy for obesity. Methods: Primary literature was searched using SciFinder and Google Scholar. Patents were searched using the European Patent Office and SciFinder. Results/conclusion: Several classes of small molecule GHS-R antagonists have been reported to be efficacious in rodent models for weight loss but none has advanced to human clinical trials. Antagonizing GHS-R as a therapy targeting the general obese population is a challenging strategy.     

树突状细胞联合细胞因子诱导的杀伤细胞体外抗肺癌的实验研究

目的探讨树突状细胞（DC）联合细胞因子诱导的杀伤细胞（CIK）在体内外能否有效的抗肺癌。方法正常人外周血单个核细胞经不同细胞因子作用后培养成DC和CIK细胞,单个核细胞经贴附塑料平皿获得单核细胞,加入GM—CSF,IL-4及TNF—a诱导获得DC,悬浮细胞加入IFN—g,24h后IL-1a,IL-2及CD3McAb诱导获得CIK,将A549制备成肿瘤细胞冻融物．作为肿瘤抗原刺激DC,并将DC与CIK细胞联合培养（DC＋CIK,负载抗原的DC＋CIK）,以CIK细胞单独培养作为对照。CytotoxicityTOX96体外杀伤实验测定体外细胞毒活性。结果在培养的第15d．与负载抗原的DC共同培养的CIK与CIK细胞单独培养细胞相比,增殖速率明显提高[（23．4±2．3）倍vs（16．7±2．7）倍,P〈0．05],CD3＋CD56＋细胞表达水平也明显提高,[（64．3±3．6）％vs（43．9±2．1）％,P〈0．05],同时负载抗原的DC的CIK对A549细胞的体外下细胞毒活性增强。结论DC与CIK细胞共培养后可提高CIK细胞的增殖速率,提高CIK细胞表型的表达水平,增强CIK抗肺癌的活性．将来可作为一种临床有效的过继免疫治疗策略。     

Role of the endothelin axis and its antagonists in the treatment of cancer

The endothelins (ET) are a group of proteins that act through G-protein coupled receptors. Endothelin-1 (ET-1) was initially identified as a potent vasoconstrictor and dysregulation of the ET axis contributes to pathological processes responsible for cardiovascular disease states. More recently, the ET axis, in particular ET-1 acting through the endothelin A receptor (ET(A) ), has been implicated in the development of several cancers through activation of pathways involved in cell proliferation, migration, invasion, epithelial-mesenchymal transition, osteogenesis and angiogenesis. The endothelin B receptor (ET(B) ) may counter tumour progression by promoting apoptosis and clearing ET-1; however, it has recently been implicated in the development of some tumour types including melanomas and oligodendrogliomas. Here, we review emerging preclinical and clinical data outlining the role of the ET axis in cancer, and its antagonism as an attractive and challenging approach to improve clinical cancer management. Clinical data of ET(A) antagonists in patients with prostate cancer are encouraging and provide promise for new ET(A) antagonist-based treatment strategies. Given the unexpected opportunities to affect pleiotrophic tumorigenic signals by targeting ET(A)-mediated pathways in a number of cancers, the evaluation of ET-targeted therapy in cancer warrants further investigation.     

Abarelix: the first gonadotrophin-releasing hormone antagonist for the treatment of prostate cancer

The high incidence of prostate cancer makes it a major healthcare problem and the second leading cancer-related cause of death among men in developed countries. The hormonal treatment of prostate cancer is indicated for the palliation of symptomatic and metastatic disease in older patients, and as neoadjuvant treatment of different modalities of radiotherapy. This hormonal treatment is based on the study conducted by Huggins in 1940 and consists of androgen suppression. Since the clinical availability of the first luteinising hormone-releasing hormone (LHRH) agonist, no significant improvement has been made in the field of medical castration. Taking these data into consideration, the recent approval of abarelix by the FDA, the first gonadotrophin-releasing hormone (GnRH) antagonist, appears to be promising news. The pharmacology of the molecule and the clinical studies that led to FDA approval will be reviewed. The place of GnRH antagonists in the treatment modalities of prostate cancer will then be discussed.     

树突状细胞联合细胞因子诱导的杀伤细胞体内外抗肺癌的实验效应研究

目的探讨树突状细胞（DC）联合细胞因子诱导的杀伤细胞（CIK）在体内外对肺癌的影响。方法正常人外周血单个核细胞经不同细胞因子作用后培养成DC和CIK细胞,单个核细胞经贴附塑料平皿获得单核细胞,加入GM-CSF,IL-4及TNF-a诱导获得DC,悬浮细胞加入IFN-g,24h后IL-1a,IL-2及CD3McAb诱导获得CIK,将A549制备成肿瘤细胞冻融物,作为肿瘤抗原刺激DC,并将DC与CIK细胞联合培养（DC＋CIK,负载抗原的DC＋CIK）,以CIK细胞单独培养作为对照。用流式细胞仪分析细胞表型,自体混合淋巴细胞反应和异体混合淋巴细胞反应检测其刺激T细胞的的增殖活性。Cytotoxicity TOX96体外杀伤实验测定体外细胞毒活性,同时用A549细胞系建立荷瘤裸鼠模型研究其体内抗肺癌活性。结果在培养的第15d,与负载抗原的DC共同培养的CIK与CIK细胞单独培养细胞相比,增殖速率明显提高[（23.4±2.3）倍vs（16.7±2.7）倍,P〈0.05],CD3＋CD56＋细胞表达水平也明显提高,[（64.3±3.6）%vs（43.9±2.1）%,P〈0.05],同时负载抗原的DC的CIK对A549细胞的体外下细胞毒活性增强,体内实验显示,与单独培养的CIK细胞相比,与负载抗原的DC共同培养的CIK,可明显抑制接种瘤细胞的裸鼠成瘤率,DC＋CIK与负载抗原的DC＋CIK在抗六效应上没有明显差异。结论 DC与CIK细胞共培养后可提高CIK细胞的增殖速率,提高CIK细胞表型的表达水平,增强CIK体内抗肺癌的活性。将来可作为一种临床有效的过继免疫治疗策略。     

Identification of biosynthetic intermediates of teaghrelins and teaghrelin-like compounds in oolong teas,and their molecular docking to the ghrelin receptor

Teaghrelins are unique acylated flavonoid tetraglycosides found in Chin-shin oolong tea, and have been demonstrated to be promising oral ghrelin analogues. The biosynthetic pathway of teaghrelins from quercetin-3-O-rutinoside (rutin) or kaempferol-3-O-rutinoside (nicotiflorin) was proposed to comprise three enzymatic steps according to the identification of putative intermediates in Chin-shin oolong tea. In addition to the two known teaghrelins in Chin-shin oolong tea, four teaghrelin-like compounds with different attachments of glycosides were identified in various oolong teas. Molecular modeling and docking were used to evaluate theoretically whether the putative biosynthetic intermediates of teaghrelins and the four teaghrelin-like compounds could be potential candidates of ghrelin analogues. The results showed that the attachment of a coumaroyl group was crucial for these tea compounds to bind to the ghrelin receptor. However, the additional attachment of a rhamnosyl glycoside to the flavonoid backbone of teaghrelin-like compounds at C-7 significantly reduced their binding affinity with the ghrelin receptor.     

番茄红素对人前列腺癌细胞（DU145）生长抑制的离体和整体水平研究

目的：研究番茄红素对人前列腺癌细胞DU145的影响。方法：体外培养的人前列腺癌细胞DU145经番茄红素作用后用苔盼蓝色进行活细胞计数,绘制生长曲线并计算抑制率;应用流式细胞仪分析番茄红素对DU145细胞周期和凋亡的影响。制作裸鼠人前列腺癌模型,观察番茄红素在整体动物上的作用。结果：番茄红素可以明显抑制DU145细胞的生长,且呈剂量－效应和时间－效应关系。10、20、40μmol/L处理细胞7d后,细胞增长的抑制率分别为10．76％、92．90％99．11％（P＜0．01）。裸鼠人前列腺癌模型上的研究显示,6％番茄红素油树脂可显著抑制腺瘤的生长、300mg/kg剂量组腺癌抑制率可达到75．76％（P＜0．05）。用经番茄红素处理过的细胞致瘤力消失。流式细胞仪分析显示番茄红素影响该细胞周期并引起细胞凋亡,凋亡率高达42．42％。结论：番茄红素可以在离体细胞和整体动物水平明显抑制雄激素非依赖型人前列腺癌DU145,其抑制机制可能是通过影响人前列腺癌细胞的生长周期和诱导其凋亡而实现。     

Darolutamide (ODM-201) for the treatment of prostate cancer

Introduction: Androgen deprivation therapy (ADT) is a mainstay initial treatment for advanced hormone-sensitive prostate cancer (HSPC), but disease progression to castration-resistant prostate cancer (CRPC) invariably occurs when patients do not succumb to another disease or comorbidity. Recognition that the androgen receptor (AR) axis continues to drive disease progression has led to the development of several AR-directed approved agents, including abiraterone acetate and enzalutamide. An investigational agent, darolutamide (ODM-201, BAY-1841788), has completed early-phase clinical trials, and two global phase III trials are currently accruing patients.

Areas covered: The unmet clinical need, pharmacokinetics, preclinical development, and clinical efficacy and safety of darolutamide for the treatment of advanced prostate cancer are reviewed. The design of two ongoing phase III trials (ARAMIS and ARASENS) of darolutamide in men with non-metastatic CRPC and metastatic HSPC, respectively, are also discussed.

Expert opinion: Darolutamide is an oral, investigational, high-affinity AR antagonist which has activity against known AR mutants that confer resistance to other second-generation antiandrogens, has minimal blood–brain barrier penetration, and does not significantly increase serum testosterone. These features may offer potential advantages over the second-generation antiandrogens. In the phase I/II ARADES trial, darolutamide demonstrated promising antitumor activity and a favorable safety profile in men with metastatic CRPC.     

油菜花粉脂溶性提取物对人前列腺癌细胞PC-3的体外生长抑制研究

目的探讨油菜花粉脂溶性提取物(BPE)对体外培养的人前列腺癌细胞PC-3的杀伤效应。方法光镜观察用药前后PC-3细胞形态,透射电镜观察细胞凋亡的形态变化;流式细胞术检测BPE对PC-3细胞凋亡的影响;反转录-聚合酶链反应(RT-PCR)检测bax/bcl-2基因表达。结果 BPE对PC-3细胞增殖有抑制作用,PC-3细胞随着BPE浓度的增高其活细胞数下降,呈负相关。透射电镜下可见:核结构破坏,染色质稀疏,有些凝聚成团。随着时间的延长,细胞凋亡率上升,BPE与多西他赛能一同促进PC-3细胞发生凋亡。BPE的作用效应不是通过bax/bcl-2基因。结论 BPE可抑制人前列腺癌细胞PC-3的增殖,诱导细胞的凋亡。     

用于治疗前列腺癌的雄激素受体拮抗剂

前列腺癌是男性泌尿生殖系统常见的恶性肿瘤,雄激素受体与前列腺细胞发育和前列腺癌发生密切相关,有可能成为治疗前列腺癌的重要靶点。对前列腺癌进行去势治疗后却常见发生更为凶险的去势抵抗性前列腺癌,因此,寻找对去势抵抗性前列腺癌有效的雄激素受体拮抗剂是当今研究热点。综述雄激素受体拮抗剂的作用机制及研究进展。