N-Acetylated alpha-linked acidic dipeptidase expressed in rat adipocytes is localized in the insulin-responsive glucose transporter (GLUT4) intracellular compartments and involved in the insulin-stimulated GLUT4 recruitment

The GLUT4-containing vesicles purified from rat adipocyte contain many protein species of unknown identity, some of which are likely to play a critical role in the trafficking of GLUT4. Presently, we describe an 85-kDa protein in GLUT4-vesicles of rat adipocytes as a potential GLUT4 traffic regulatory protein. MALDI-TOF MS, RT-PCR, gene cloning, protein sequence analysis, and immunoreactivity assay have identified this protein as N-acetylated alpha-linked acidic dipeptidase (NAALADase) expressed in rat adipocytes. NAALADase in rat adipocytes was mostly membrane-associated and colocalized in discrete GLUT4-compartments with enrichment in putative GLUT4-sorting endosomes (G4G(L)). Total cell lysates of adipocytes exhibited NAALADase activity. Next, we treated rat adipocytes with 2-[phosphonomethy]pentanedionic acid (2-PMPA), a potent NAALADase inhibitor, and studied its effect on the distribution of GLUT4 and 3-O-methyl glucose (3OMG) flux. In 2-PMPA-treated adipocytes, there was a significant reduction (by 40%) in the insulin-stimulated GLUT4 translocation to the plasma membrane. The 3OMG flux in insulin-stimulated adipocytes was also delayed (51% of control) by 2-PMPA treatment, indicating that 2-PMPA impairs insulin-stimulated GLUT4 recruitment and the uptake of glucose. It is suggested that NAALADase may function as a regulator required for the insulin-stimulated GLUT4 vesicle movement and/or its exocytosis, thus may regulate insulin-induced GLUT4 recruitment in rat adipocytes.     

Synthesis and biological evaluation of Doxorubicin-containing conjugate targeting PSMA

Prostate-specific membrane antigen (PSMA), also known as glutamate carboxypeptidase II (GCPII), has recently emerged as a prominent biomarker of prostate cancer (PC) and as an attractive protein trap for drug targeting. At the present time, several drugs and molecular diagnostic tools conjugated with selective PSMA ligands are actively evaluated in different preclinical and clinical trials. In the current work, we discuss design, synthesis and a preliminary biological evaluation of PSMA-specific small-molecule carrier equipped by Doxorubicin (Dox). We have introduced an unstable azo-linker between Dox and the carrier hence the designed compound does release the active substance inside cancer cells thereby providing a relatively high Dox concentration in nuclei and a relevant cytotoxic effect. In contrast, we have also synthesized a similar conjugate with a stable amide linker and it did not release the drug at all. This compound was predominantly accumulated in cytoplasm and did not cause cell death. Preliminary in vivo evaluation has showed good efficiency for the degradable conjugate against PC3-PIP(PSMA⁺)-containing xenograft mine. Thus, we have demonstrated that the conjugate can be used as a template to design novel analogues with improved targeting, anticancer activity and lower rate of potential side effects. 3D molecular docking study has also been performed to elucidate the underlying mechanism of binding and to further optimization of the linker area for improving the target affinity.     

PSA在前列腺增生和前列腺癌中的诊断价值

目的：探讨临床上检测前列腺特异性抗原（PSA）的变化情况对前列腺增生和前列腺癌等疾病的诊断价值。方法：采用回顾性分析的方法,选取2010年6月至2012年4月在我院泌尿科接受治疗的前列腺增生患者64例定义为前列腺增生组（BPH）,前列腺癌患者83例定义为前列腺癌组（PCa）,另选取同期接受体检的健康人群137例作为对照组。分别检测三组患者入院时的游离前列腺特异性抗原和总前列腺特异性抗原的水平变化情况。对比并分析三组检测结果。结果：经检测,前列腺增生患者的血清总PSA明显高于对照组健康人群的正常值,而前列腺癌患者的血清总PSA比前列腺增生患者增高的更为明显。对照组游离PSA为（2．78±0．94）ng／mL,总PSA（1．05±0．57）ng／mL,游离PSA与总PSA的比值为,（0．38±0．61）;前列腺增生患者游离PSA为（6．36＋3．24）ng／mL,总PSA为（1．64±0．76）ng／mL,游离PSA与总PSA的比值为（0．26±0．23）;前列腺癌患者游离PSA为（12．42±4．97）ng／mL,总PSA为（1．44±0．78）ng／mL,游离PSA与总PSA的比值为（0．12±0．16）。组间比较差异明显,具有统计学意义（P〈0．05）。结论：对患者的PSA进行检测,对前列腺增生和前列腺癌的诊断具有良好的辅助作用和,临床价值。     

In vitro Method to Observe E-selectin-mediated Interactions Between Prostate Circulating Tumor Cells Derived From Patients and Human Endothelial Cells

Metastasis is a process in which tumor cells shed from the primary tumor intravasate blood vascular and lymphatic system, thereby, gaining access to extravasate and form a secondary niche. The extravasation of tumor cells from the blood vascular system can be studied using endothelial cells (ECs) and tumor cells obtained from different cell lines. Initial studies were conducted using static conditions but it has been well documented that ECs behave differently under physiological flow conditions. Therefore, different flow chamber assemblies are currently being used to studying cancer cell interactions with ECs. Current flow chamber assemblies offer reproducible results using either different cell lines or fluid at different shear stress conditions. However, to observe and study interactions with rare cells such as circulating tumor cells (CTCs), certain changes are required to be made to the conventional flow chamber assembly. CTCs are a rare cell population among millions of blood cells. Consequently, it is difficult to obtain a pure population of CTCs. Contamination of CTCs with different types of cells normally found in the circulation is inevitable using present enrichment or depletion techniques. In the present report, we describe a unique method to fluorescently label circulating prostate cancer cells and study their interactions with ECs in a self-assembled flow chamber system. This technique can be further applied to observe interactions between prostate CTCs and any protein of interest.     

前列腺癌患者前列腺体积与肿瘤分级之间关系探讨

目的:探讨国人前列腺癌患者前列腺体积与肿瘤分级之间的关系。方法:回顾我院及武汉大学人民医院2005年1月-2011年10月70例确诊为前列腺癌并行根治性前列腺切除术(RP)患者的临床病理资料,采用SPSS13.0软件总结并分析前列腺癌患者前列腺体积与肿瘤分级之间的关系。结果:经直肠前列腺穿刺活检获得肿瘤病理分级与根治性前列切除术获得最终病理分级具有显著差异(P=0.003);在活检及根治性前列腺切除标本中,前列腺体积与高级别肿瘤发生率均呈负相关(P<0.05);小前列腺与阳性手术切缘、前列腺外侵犯及高级别肿瘤在单变量分析中具有相关性(P<0.05),而与精囊腺侵犯及淋巴结侵犯则无相关性(P>0.05);在校正了年龄、体重指数及术前前列腺特异性抗原水平后,前列腺体积与阳性手术切缘、前列腺外侵犯、精囊腺侵犯及高级别肿瘤发生率均呈负相关(OR<1,P<0.05),而与淋巴结侵犯则无相关性(P>0.05)。结论:前列腺体积是高级别前列腺癌的重要预测因子,利用其对高级别肿瘤风险的预测能力可帮助选择最佳治疗方案并进一步提高治疗效果。     

超声弹性成像技术在前列腺癌诊断中的应用

超声弹性成像技术是一项新兴的超声成像方法,可通过分析不同组织间机械组织差异区分组织软硬度,早期主要应用于乳腺、甲状腺结节的区分和定性。随着科技的进步和医疗水平的提高,目前弹性成像技术可在二维声像图的基础上,对感兴趣区域进行定性诊断和定量分析,已逐步应用于医学各领域相关研究和临床疾病的鉴别诊断。本文介绍了弹性成像的基本原理和目前弹性成像技术在医学领域中的相关应用,叙述了前列腺癌的发展趋势和目前常用的筛查、诊断方法,详细阐述了弹性成像技术在前列腺癌诊断中的应用方法和现状,分析总结了弹性成像技术在前列腺癌诊断中的应用价值。运用弹性成像技术无创、经济便捷、实时动态、可重复性好等优点,联合前列腺癌相关筛查、诊断检查,可有效帮助早期诊断前列腺癌,减少前列腺穿刺术的针数,提高前列腺癌的检出率。     

TGF-beta在前列腺癌骨转移中的研究进展

进展期前列腺癌多会发生骨转移,导致患者骨质破坏甚至死亡。前列腺癌发生骨转移的机制目前尚未研究清楚。既往多认 为是因为前列腺癌细胞表面携带者容易在骨环境中生长的表型。但是目前多认为是肿瘤细胞与骨骼微环境之间的相互作用导致的结果。它们之间是通过细胞因子来传递信息。在众多因子中,TGF-beta对前列腺癌骨转移灶中的各种细胞都起着重要作用。研究表明在体外实验中TGF-beta的作用极易受到细胞生长环境的影响,表现出不同的功能。这提示着TGF-beta信号通路和其他信号通路之间存在非常强的交互作用。本文的重点在于对TGF-beta在前列腺癌骨转移中的作用研究进展进行综述,阐述TGF-beta对转移灶中不同细胞的作用,为今后肿瘤的治疗研究寻找一个好的方向。     

一个以前列腺特异抗原密度和穿刺评分为基础的中国人群中前列腺癌根治术后病理升级预测模型的建立

目的：分析前列腺癌根治术后病理得分较穿刺得分增加的原因,并建立一个可以预测中国人群中前列腺癌根治术后病理升级的模型。方法：以2008 年8 月至2013 年12 月在我院泌尿科行前列腺癌根治性切除术的264例患者的临床资料为基础,根据术前和术后患者病理得分的变化将其分为升级组和未升级组。运用单因素和多因素logistic 回归分析病理升级的原因,并通过多因素回归系数建立预测病理升级的诺模图。结果：264 例患者中,共238 例最终纳入统计分析,多因素logistic 回归分析显示前列腺特异抗原密度(0R=3.854,P=0.001 )和穿刺Gleason(≤ 6)评分是中国人群中前列腺癌根治术后病理升级的独立危险因素。前列腺特异抗原密度和穿刺得分的ROC 最佳截断取值为0.37 ng/ml 2和8 分。运用上述两个变量建立了一个可用于预测病理升级的诺模图。结论：前列腺特异抗原密度和穿刺Gleason 评分是预测中国人群中前列腺癌根治术后病理升级的独立危险因素,本研究所得的诺模图可以很好地预测前列腺癌根治术后的病理升级。     

miR-182在前列腺癌组织中的表达及其对前列腺癌细胞增殖和凋亡的影响

目的:观察前列腺癌组织及不同前列腺癌细胞系中miR-182的表达,并探讨下调其表达对前列腺癌细胞增殖和凋亡的影响及机制。方法:采用实时荧光定量PCR(q RT-PCR)检测30例前列腺癌组织和30例相应的癌旁组织以及前列腺正常上皮RWPE-1细胞、前列腺癌PC-3、LNCa P和DU145细胞中miR-182的表达,进一步采用Lipfectamine 2000脂质体转染miRNA-182 inhibitor和阴性对照miRNA于PC-3细胞后,通过噻唑蓝(MTT)比色法检测细胞增殖情况,流式细胞术检测细胞凋亡率,免疫印迹(Western blot)法检测转录因子FOXO1、血管内皮生长因子(VEGF)和抑癌基因p53蛋白的表达。结果:miR-182在前列腺癌组织中的表达明显高于癌旁组织(P0.05);miR-182在前列腺癌细胞系PC-3、LNCa P和DU145中的表达均高于前列腺正常上皮细胞RWPE-1(P0.05),其中PC-3细胞中miR-182表达水平最高。转染miRNA-182 inhibitor至PC-3细胞成功下调miR-182表达后,细胞的增殖能力明显受到抑制,细胞凋亡能力明显增强,FOXO1表达水平显著升高,VEGF和p53的表达明显降低,差异均具有统计学意义(P0.05)。结论:miR-182在前列腺癌组织及细胞中呈高表达,下调miR-182的表达可能通过增加FOXO1的表达并减少VEGF和p53的表达,抑制前列腺癌细胞增殖并诱导细胞凋亡。     

腹腔镜前列腺癌根治术治疗前列腺癌的疗效及对患者血清激素水平的影响

目的:分析腹腔镜前列腺癌根治术治疗前列腺癌的疗效及其对患者血清激素水平的影响。方法:选择86例前列腺癌患者为研究对象并按抽签法随机分为对照组与观察组,每组43例。对照组行开放前列腺癌根治术治疗,观察组采用腹腔镜前列腺癌根治术,比较两组手术指标,手术前后血清促卵泡激素(FSH)、黄体生成素(LH)、双氢睾酮(DHT)、游离睾酮(FT)、总睾酮(T)、前列腺特异性抗原(T-PSA、F-PSA),CD3~+、CD4~+、CD8~+、CD4~+/CD8~+水平的变化及术后并发症的发生情况。结果:观察组手术时间明显长于对照组,失血量、肠功能恢复时间、住院时间、疼痛评分均明显低于或短于对照组(P0.05)。两组治疗前后血清FSH、LH、DHT、FT、T、T-PSA、F-PSA比较差异均无统计学意义(P0.05)。观察组CD3~+、CD4~+、CD4~+/CD8~+显著高于对照组(P0.05),CD8~+低于对照组(P0.05)。观察组术后并发症的发生率明显低于对照组(P0.05)。结论:腹腔镜前列腺癌根治术可起到与开放手术相似的控瘤效果,提高血清雄激素水平和患者的免疫功能,控制肿瘤进展,且安全性更高。     

PSA相关指标对前列腺癌诊断价值的评价及比较研究

目的:探讨血清总PSA(TPSA)、F/TPSA和PSA密度(PSAD)在前列腺癌(PCa)诊断中的价值,寻找更准确的前列腺癌诊断指标。方法:采用化学发光免疫分析法检测前列腺癌患者(60例)和前列腺增生患者(240例)的血清PSA水平,通过B超测定患者前列腺的体积,计算PSAD,运用ROC曲线评价和比较血清总PSA(TPSA)、F/TPSA和PSAD诊断前列腺癌的准确性和特异性。结果:(1)前列腺癌患者TPSA和PSAD值均明显高于前列腺增生患者(P0.01),F/TPSA明显低于前列腺增生患者(P0.01);(2)TPSA阈值定为4 ng/ml时,诊断前列腺癌的敏感性、特异性分别为56.23%、80.10%。F/TPSA阈值定为0.15时,诊断前列腺癌的敏感性、特异性分别为88.10%、69.10%,PSAD阈值定为0.20时,诊断前列腺癌的敏感性、特异性分别为88.60%、88.30%。结论:TPSA、F/TPSA和PSAD在前列腺癌诊断中均有一定的价值,且PSAD诊断前列腺癌的敏感性、特异性优于TPSA、F/TPSA,是诊断前列腺癌更为理想的指标。     

Association of glutamate carboxypeptidase II (GCPII) haplotypes with breast and prostate cancer risk

In view of the pivotal role of glutamate carboxypeptidase II (GCPII) in carcinogenesis, its expression as prostate specific membrane antigen (PSMA) and folate hydrolase (FOLH1) may be influenced by its haplotypes, contributing to the etiology of prostate and breast cancer. To test this hypothesis, breast and prostate cancer cases and controls were subjected to whole gene screening of GCPII and correlated with plasma folate levels and PSMA expression. The impact of variants on a 3-dimensional structure of GCPII was explored by in silico studies. Six novel variations i.e. V108A, P160S, Y176H, D191V, G206R and G245S; and two known variations i.e. R190W and H475Y were identified in GCPII. All-wild haplotype and a haplotype harbouring D191V showed association with breast cancer risk while haplotypes harbouring V108A and P160S reduced the risk. Haplotypes with V108A and G245S variants showed increased risk for prostate cancer due to high PSMA expression while P160S conferred protection against prostate cancer. In silico studies suggests that P160S and R190W variants result in relaxed substrate binding facilitating either rapid catalysis or exchange of substrates and products in the active site which was substantiated by high plasma folate levels associated with these variants. On the contrary, D191V was associated with very low plasma folate levels despite having a high PSMA expression. This is the first comprehensive study examining variations in GCPII in relation to breast and prostate cancer risk. Changes in the plasma folate levels and changes in PSMA expression are associated with breast and prostate cancer risk respectively.     

姜黄素和紫杉醇联用对前列腺癌PC3裸鼠移植瘤作用的研究

目的:探讨姜黄素和半量紫杉醇联用对前列腺癌PC3裸鼠移植瘤生长增殖的影响及其机制。方法:构建人雄激素非依赖性前列腺癌细胞系PC3裸小鼠皮下移植瘤模型,随机分为溶酶对照组,姜黄素组,紫杉醇组和姜黄素+紫杉醇(半量)组(n=6)。姜黄素每隔2天腹腔注射100mg/kg,紫杉醇每3天腹腔注射10mg/kg,联合组紫杉醇减半,对照组注射药物溶剂。每6天测量计算移植瘤体积并绘制肿瘤生长曲线。药物注射30天后,取移植瘤组织标本分别进行免疫组化和定量RT-PCR,检测金属基质蛋白酶2(MMP2),增殖细胞核抗原(PCNA)蛋白及mRNA的表达。结果:移植瘤体积在24～30天时,姜黄素组和紫杉醇组肿瘤体积较对照组有不同程度的减小。姜黄素+紫杉醇(半量)组在第30天内肿瘤生长体积和紫杉醇组相近,30天后合用组肿瘤体积小于单纯紫杉醇组(P<0.05)。姜黄素和紫杉醇明显降低PC3移植瘤组织中PCNA和MMP2mRNA的表达(P<0.01)。姜黄素+紫杉醇(半量)组瘤组织中PCNA和MMP2的mRNA表达均低于紫杉醇组(P<0.05)。免疫组化染色显示PCNA和MMP2蛋白表达在治疗组均降低,和瘤组织中mRNA表达变化相一致。结论...     

A critical update on the strategies towards modulators targeting androgen receptors

Prostate cancer is the most common carcinoma of the male urinary system in developed countries. Androgen deprivation therapy has been commonly used in the treatment of prostate cancer for decades, but most patients will inevitably develop into more aggressive castration-resistant prostate cancer. Therefore, novel strategies are urgent to address this resistance mechanism. In this review, we discussed some new strategies for targeting androgen receptors through degradation pathways as potential treatments for prostate cancer.     

Radiopharmaceuticals targeting PSMA for imaging and/or therapy

Targeting the Prostate Specific Membrane Antigen (PSMA) is becoming increasingly more important for the management of prostate cancer patients at various stages. This review article describes selected radiolabelled PSMA inhibitors with optimized radiopharmaceutical properties for imaging and/or therapy.     

磁共振成像联合血清前列腺特异抗原、上皮钙黏蛋白、早期前列腺癌抗原-2诊断前列腺癌的临床价值探讨

摘要 目的：探究磁共振成像（MRI）联合血清前列腺特异抗原（PSA）、上皮钙黏蛋白（sE-cadherin）、早期前列腺癌抗原-2（EPCA-2）诊断前列腺癌的临床价值。方法：选取潍坊市人民医院2020年1月-2021年7月期间经病理证实的50例前列腺癌患者（前列腺组）以及50例前列腺增生患者（前列腺增生组）展开回顾性研究。100例研究对象均完善了MRI检查并测定血清PSA、EPCA-2、sE-cadherin水平,分析两组患者的MRI影像学特征,比较两组患者的PSA、EPCA-2、sE-cadherin水平以及各项检查方法的诊断准确性差异。结果：前列腺癌的MRI影像学特征为病灶主要位于外周带,外周带T2W呈低信号,病变侵及包膜、膀胱及周围组织具有T1加权消失或者不对称,具有信号异常、肌肉增厚的表现,盆腔淋巴结转移具有淋巴结部分融合或增大表现;前列腺增生的MRI影像学特征为边界清晰、包膜完整并且中央带增生、不均匀信号结节;前列腺癌、前列腺增生均存在不同程度的前列腺体积增大。相比于前列腺增生组,前列腺癌组患者的PSA、sE-cadherin、EPCA-2水平明显更高（P<0.05）。MRI、PSA、sE-cadherin、EPCA-2四项联合鉴别前列腺癌、前列腺增生的诊断符合率为96.00%,明显高于四项单独诊断的88.00%、79%、81%、82%（P<0.05）。结论：MRI联合PSA、sE-cadherin、EPCA-2鉴别诊断前列腺癌的准确性较高,具有作为临床前列腺癌早期诊断指导方案的潜力。     

心理干预对老年前列腺癌去势患者术后心理状态的影响

目的:探讨围术期心理干预对老年前列腺癌去势患者心理状况及生活质量的影响。方法:选取2012年6月-2015年1月在我院接受手术治疗的老年前列腺癌患者150例,根据围术期干预方法的不同将所选患者分为研究组和对照组,每组75例。对照组患者给予常规护理干预,研究组在对照组基础上给予心理干预。分别于干预前后应用生活质量核心问卷(QLQ-C30)评价患者生活质量,应用焦虑自评量表(SAS)评价患者焦虑情况,应用抑郁自评量表(SDS)评价患者抑郁情况。结果:干预前,两组患者QLQ-C30、SAS和SDS评分比较均无统计学意义(P0.05);干预后,两组患者QLQ-C30、SAS和SDS评分均显著优于干预前,组内比较,差异具有统计学意义(P0.05);干预后,研究组QLQ-C30、SAS和SDS评分优于对照组,两组比较差异具有统计学意义(P0.05)。结论:围术期心理干预对老年前列腺癌去势患者具有较好的干预效果,能够显著改善患者的心理状态,提高术后生活质量。     

长链非编码RNA LINC01006对前列腺癌细胞增殖和侵袭的影响

摘要 目的：探究长链非编码RNA LINC01006对前列腺癌（prostate cancer, PCa）细胞增殖和侵袭能力的影响。方法：体外培养人前列腺正常上皮细胞系RWPE-1,人PCa细胞系LNCaP、22Rv1、PC3、C4-2b,应用实时定量PCR（qRT-PCR）技术检测上述细胞LINC01006的表达;分别通过转染小干扰RNA（siRNA）或过表达LINC01006的慢病毒载体,在LNCaP和PC3细胞中敲减LINC01006或稳定过表达LINC01006;应用CCK8、克隆形成实验检测LINC01006对PCa细胞增殖能力的影响;应用Transwell侵袭实验检测LINC01006对PCa细胞侵袭能力的影响;通过网站预测LINC01006的转录调控因子及其结合位点。结果：相较于正常前列腺上皮细胞系RWPE-1,PCa细胞系LNCaP、22Rv1、C4-2b和PC3中LINC01006表达明显升高（P<0.05）。敲减LINC01006后的PCa细胞系LNCaP和PC3的增殖和侵袭能力被显著抑制（P<0.05）,过表达LINC1006则明显促进PCa细胞系LNCaP和PC3的增殖、侵袭能力（P<0.05）。通过PROMO网站预测可见AR是LINC01006的潜在转录调控因子,通过Cistrome DB数据库发现LINC01006上游启动子区域存在AR富集;敲减、抑制AR后LNCaP细胞中LINC01006表达明显升高（P<0.05）。结论：LINC01006在PCa细胞系中呈高表达,促进PCa细胞的增殖和侵袭,其受到AR负向调控,可能在PCa发生发展和去势抵抗形成过程中发挥作用。     

五苓胶囊对前列腺癌循环miRNA-141表达的影响

目的:采用荧光定量PCR法检测前列腺癌循环miRNA-141表达,并研究五苓胶囊对其影响。方法:选取我院泌尿外科收治的前列腺癌患者20例,随机分为A、B两组,各10例;A组给予常规治疗,B组在常规治疗基础上给予加用五苓胶囊治疗。选取同期住院前列腺增生患者20例,正常对照组为同时期体检健康者20例。采用荧光定量PCR法检测正常对照组、前列腺增生组以及前列腺癌组血清miRNA-141的表达情况,以及前列腺癌组治疗前TPSA(前列腺特异性抗原)含量。结果:前列腺癌组的血清miRNA-141的表达水平为正常对照组的(11.22±8.19)倍,明显高于前列腺增生组(1.65±1.73)和正常对照组,差异有统计学意义(P0.05);前列腺增生组和正常对照组比较无明显差异(P0.05);前列腺癌组血清miRNA-141的表达水平与TPSA水平呈正相关关系,相关系数r=0.753,有统计学意义(P0.05)。治疗后,前列腺癌A组与B组的血清miRNA-141的表达水平均有所改善,与A组(5.82±4.51)相比较,B组的miRNA-141的表达水平(1.93±0.67)明显降低,差异有统计学意义(P0.05)。结论:前列腺癌患者血清中miRNA-141表达水平明显升高,且与TPSA水平呈正相关关系,五苓胶囊能够降低miRNA-141表达。