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。     

血糖与前列腺癌患者的关系

目的:探讨前列腺活检患者的血糖与前列腺癌患者的关系。方法:前瞻性收集416例初次经直肠超声引导下前列腺穿刺活检患者的血糖、前列腺特异性抗原(PSA)和Gleason评分等临床资料,所有患者以血糖浓度6.1 mmol/L为界分成两组,比较高血糖组和正常血糖组前列腺癌检出率和Gleason评分的差异。结果:416例前列腺活检患者中,检出前列腺癌165例,高血糖组38例(40.00%),正常血糖组127例(39.56%),差异无统计学意义(P0.05);低级别前列腺癌(Gleason7分)患者的构成比分别为0.184、0.071,差异有统计学意义(P0.05),Spearman等级相关分析显示前列腺癌患者的血糖值与Gleason评分呈负相关(r=-0.228,P0.05)。结论:血糖对前列腺活检患者中前列腺癌检出率没有影响,但提高了低级别前列腺癌患者的构成比,血糖是影响前列腺癌Gleason评分的一个独立因素。     

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。     

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。     

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。     

The specificity of interphase FISH translocation probes in formalin fixed paraffin embedded tissue sections is readily assessed using automated staining and scoring of tissue microarrays constructed from murine xenografts

Implementation of interphase fluorescence in situ hybridization (FISH) assays in the clinical laboratory requires validation against established methods. Validation tools in common use include exchange of consecutive sections with another institution that has already established the FISH assay, comparison with conventional banded metaphase cytogenetics, confirmation of specificity using probed normal metaphases, consecutive paraffin sections of a validation set tested by a reference laboratory, and specificity assessment against well characterized cell lines. We have investigated the feasibility of using tissue microarrays (TMA) constructed from murine xenografts as a preliminary specificity-screening tool for validation of interphase FISH assays. Cell lines currently in use for FISH controls are used to generate xenografts in SCID mice which are fixed in formalin and paraffin embedded. A TMA is constructed using duplicate donor cores from the xenograft blocks. Xenografts used represent a wide range of translocations used routinely for formalin fixed paraffin embedded sections evaluated by FISH. Probe cocktails (Abbott-Vysis), for several non-random translocations associated with hematologic neoplasms and soft tissue sarcomas have been used in this manner. On-line deparaffinization, cell conditioning, and prehybridization steps are automated using a staining workstation (Ventana Discovery XT); hybridization and stringency washes are performed manually offline. FISH-probed TMAs are tracked using a Metasystems image scanner and analyzed using classifiers specifically developed for each molecular abnormality. FISH results for each xenograft in the TMA correspond exactly to the genotype previously established for the parent cell line from which the xenograft was prepared. Moderate complexity tissue microarrays constructed from murine xenografts are excellent validation tools for initial assessment of interphase FISH probe specificity.     

帕比司他逆转前列腺癌细胞hepaCAM基因表达机制研究

目的:研究帕比司他(Panobinostat)逆转抑癌基因肝细胞粘附分子(hepatocyte cell adhesion molecule,hepaCAM)表达,协同hepaCAM抑制前列腺癌(prostate cancer,PCa)细胞生长。方法:不同浓度帕比司他作用于体外培养的PC3细胞,首先采用MTT法检测帕比司他对细胞增殖的影响,RT-PCR和Western blot法检测hepaCAM、组蛋白去乙酰化酶(histone deacetylases, HDACs)以及乙酰化组蛋白H3 赖氨酸9(Ac-H3K9)的表达变化。接着用不同因素处理细胞,MTT法检测细胞增殖活性,流式细胞术检测细胞周期改变,RT-PCR和Western blot法检测细胞周期调节因子cyclinD1 和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的基因表达。结果:帕比司他抑制PC3细胞生长与作用浓度增加和作用时间呈正相关, hepaCAM mRNA、蛋白以及细胞核中Ac-H3K9表达随帕比司他浓度升高而增高,HDAC1、 HDAC3、 HDAC4 mRNA和蛋白的表达随浓度升高而降低;单独过表达hepaCAM腺病毒和单独使用帕比司他组PC3细胞生长明显受到抑制且随作用时间延长抑制率增加,两者联用更为明显,差异有统计学意义(P < 0.05);与单独hepaCAM 腺病毒组和帕比司他组相比,两者联用S期细胞比例明显增高,差异有统计学意义(P < 0.05),且可进一步下调cyclinD1、 PCNA mRNA和蛋白的表达,差异有统计学意义(P < 0.05)。结论:帕比司他可通过抑制HDACs活性,加强PCa PC3细胞组蛋白H3 N-端的赖氨酸残基乙酰化,逆转hepaCAM表达;hepaCAM腺病毒和帕比司他联用可通过阻滞细胞周期于S期协同抑制PC3 细胞生长,作用机制可能与下调cyclinD1 和PCNA 的表达有关。这对揭示抑癌基因hepaCAM在肿瘤缺失的原因及为将帕比司他应用于临床治疗肿瘤提供了科学支持。     

嵌合分子(DHT-PROTAC)在前列腺癌研究中的进展

前列腺癌多发于老年男性,已成为老年男性常见肿瘤之一。内分泌治疗目前是晚期前列腺癌主要治疗方法,但仍避免不了前列腺癌最终进展成激素非依赖性前列腺癌,导致内分泌治疗的失败。当前,对前列腺癌细胞株的AR表达的研究,主要集中在DNA水平及mRNA水平,而对AR蛋白翻译后调控的研究较少。近些年来,嵌合分子(DHT-PROTAC)是基于蛋白水平,调控AR蛋白的表达,成为研究前列腺癌转归新的热点。DHT-PROTAC是一种新型人工合成的异型双功能小分子;这种小分子是DHT与泛素连接酶E3识别基团的嵌合体,它不仅能与AR结合,而且能在结合后,诱导AR的泛素化,从而通过泛素-蛋白酶途径降解AR;本文介绍了嵌合分子的作用原理,回顾了近些年前列腺癌的治疗进展,分析了嵌合分子将来在前列腺癌治疗中的应用前景。     

Proteomic Profiling of Androgen-independent Prostate Cancer Cell Lines Reveals a Role for Protein S during the Development of High Grade and Castration-resistant Prostate Cancer

Androgen deprivation constitutes the principal therapy for advanced and metastatic prostate cancers. However, this therapeutic intervention usually results in the transition to a more aggressive androgen-independent prostate cancer. The elucidation of molecular alterations during the progression to androgen independence is an integral step toward discovering more effective targeted therapies. With respect to identifying crucial mediators of this transition, we compared the proteomes of androgen-independent (PC3, DU145, PPC1, LNCaP-SF, and 22Rv1) and androgen-dependent (LNCaP and VCaP) and/or normal prostate epithelial (RWPE) cell lines using mass spectrometry. We identified more than 100 proteins that were differentially secreted in the androgen-independent cell lines. Of these, Protein S (PROS1) was elevated in the secretomes of all of the androgen-independent prostate cancer cell lines, with no detectable secretion in normal and androgen-dependent cell lines. Using quantitative PCR, we observed significantly higher (p < 0.05) tissue expression levels of PROS1 in prostate cancer samples, further indicating its importance in prostate cancer progression. Similarly, immunohistochemistry analysis revealed elevation of PROS1 in high grade prostate cancer (Gleason grade ≥8), and further elevation in castration-resistant metastatic prostate cancer lesions. We also observed its significant (p < 0.05) elevation in high grade prostate cancer seminal plasma samples. Taken together, these results show that PROS1 is elevated in high grade and castration-resistant prostate cancer and could serve as a potential biomarker of aggressive disease.     

TRF1、TRF2和POT1基因mRNA在前列腺癌组织中的表达

目的:探究端粒重复序列结合蛋白质1(TRF1)、TRF2和端粒保护蛋白(POT1)基因mRNA在前列腺癌(PCa)组织中的表达。方法:收集46例PCa患者肿瘤中心组织(中心组织组)和35例前列腺增生(BPH)患者BPH组织(BPH组织组),提取总RNA,逆转录成cDNA,采用定量PCR测定TRF1、TRF2和POT1在中心组织组和BPH组织基因mRNA的表达,采用基因mRNA所得CT值与β-actin mRNA所得CT值之差(△CT值)表示,并进行差异性分析。结果:中心组织组TRF1的△CT值高于BPH组织组,差异有统计学意义(Z=-3.469,P=0.001);TRF2的△CT值分别为8.49(5.75,10.21)和8.16(6.28,9.75),差异无统计学意义(Z=-1.719,P=0.086);中心组织组POT1的△CT值高于BPH组织组,差异有统计学意义(t=-18.48,P=0.000)。结论:TRF1和POT1在PCa肿瘤组织中的表达升高,说明TRF1和POT1的高表达可能与PCa的发生和发展有关,但TRF2在PCa肿瘤中心组织和BPH组织中的表达没有表现出差异,有待进一步研究。     

Anti-Oxidants from Green Tea and Pomegranate for Chemoprevention of Prostate Cancer

Among males, prostate cancer has become the second leading cause of cancer-related deaths in North America, with similar trends in many Western and developing countries. One way to control prostate cancer is through chemoprevention, which refers to the administration of synthetic or naturally occurring agents to block, reverse, or delay the process of carcinogenesis. For a variety of reasons, the most important of which is human acceptance, for chemopreventive intervention, naturally occurring diet-based agents are preferred. Prostate cancer is an ideal candidate disease for chemopreventive intervention, because it grows very slowly, likely for decades, before symptoms arise and a diagnosis is finally established, it has a long latency period, and it is typically diagnosed in men >50 years of age. Most chemopreventive agents are antioxidant in nature. We have been defining the usefulness of dietary anti-oxidants for chemoprevention of prostate and other cancers. It is increasingly appreciated that some of these dietary anti-oxidants are nature’s gift molecules endowed with cancer preventive and therapeutic properties. This review will focus on prostate cancer chemopreventive effects of polyphenolic anti-oxidants derived from green tea and pomegranate. It is a challenge to custom-tailor these gift molecules as cocktails in concentrations that can easily be consumed by humans for delaying prostate and other cancers.     

骨形态发生蛋白7 在前列腺癌中的表达

目的:探讨骨形态发生蛋白( BMP-7)在前列腺癌组织中的表达及其与临床分期之间的关系.方法:应用免疫印迹法检测30例前列腺癌患者及30例前列腺良性增生患者前列腺组织中BMP-7的表达情况.结果:前列腺癌组织中BMP-7的表达显著高于前列腺良性增生组织,且BMP-7的表达随前列腺癌的临床分期、Gleason分级增高而增加.结论:BMP-7在前列腺癌中的表达明显增高,其表达量与临床分期相关,前列腺癌组织中BMP-7的表达增高提示预后不佳.     

GPRC6A过表达前列腺癌细胞株构建及EMT相关研究

目的:构建GPRC6A基因过表达的前列腺癌LncapC4-2细胞株,检测细胞迁移和侵袭能力改变及EMT相关基因表达,为研究GPRC6A介导的EMT在前列腺癌发生发展过程中的作用奠定基础。方法:构建过表达GPRC6A的慢病毒载体pCDH-GPRC6A,人胚肾293T细胞包被病毒并感染Lncap C4-2细胞株,Puromycin筛选获得稳定过表达GPRC6A的细胞株。RT-PCR和Western blot验证细胞GPRC6A的过表达情况。划痕和transwell实验检测细胞迁移和侵袭能力,qPCR检测EMT相关基因表达。结果:成功构建GPRC6A表达的慢病毒载体,RT-PCR和Western blot检测LncapC4-2 GPRC6A过表达细胞株中GPRC6A的mRNA和蛋白质表达增高,与对照相比较,在过表达GPRC6A的LncapC4-2细胞中,细胞迁移和侵袭能力增强,并且EMT相关基因E-cadherin表达降低而SNAIL表达增高。结论:成功构建过表达的GPRC6A的前列腺癌细胞株,肿瘤细胞中的GPRC6A可能通过增强细胞迁移和侵袭能力并促进细胞EMT而参与前列腺癌发生发展过程。     

前列腺癌细胞系中环状RNA circRNA-1565的表达及鉴定

目的:探讨前列腺癌细胞系中的环状RNA circRNA-1565的表达及鉴定。方法:培养前列腺正常上皮细胞(RWPE-1)、4种前列腺癌细胞(22RV1、LNCap、PC-3、DU145),提取总RNA,采用RT-PCR检测不同细胞系中circRNA-1565的表达,并对其进行成环性验证及细胞内亚定位实验,进行差异比较。结果:circRNA-1565在正常前列腺细胞系(RWPE-1)中表达量极低,在转移性前列腺癌细胞系中高表达,在非转移性前列腺癌细胞系中低表达。且circRNA-1565是一条主要定位于细胞质内的具有有效环状结构的RNA分子。结论:circRNA-1565在不同前列腺癌细胞系中存在差异表达,可能会通过mi RNA sponge途径发挥生物学作用。     

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。     

miR-19a-3p在前列腺癌细胞侵袭和迁移中的作用

miR-19a-3p通过多种机制调节癌细胞的增殖和转移,然而,miR-19a-3p在前列腺癌转移中的生物学作用和机制尚不清楚。本研究旨在考察mir-19a-3p在前列腺癌中的表达情况,及其对细胞侵袭和迁移的影响。本研究发现,miR-19a-3p在骨转移性前列腺癌组织和前列腺癌细胞中明显下调。上调miR-19a-3p可显著抑制前列腺癌细胞的侵袭和迁移。然而,下调miR-19a-3p则会逆转上述变化。此外,本研究还发现miR-19a-3p通过靶向TGF-β信号传导的下游效应物SMAD2来抑制前列腺癌细胞中TGF-β信号传导的活性,从而抑制前列腺癌细胞的侵袭和迁移。因此,本研究表明mir-19a-3p与前列腺癌的发生发展密切相关,mir-19a-3p有望成为前列腺癌的新治疗靶点及生物标志物。     

Causes of death in men with prostate cancer: Results from the Danish Prostate Cancer Registry (DAPROCAdata)

BackgroundCurrent knowledge of the validity of registry data on prostate cancer-specific death is limited. We aimed to determine the underlying cause of death among Danish men with prostate cancer, to estimate the level of misattribution of prostate cancer death, and to examine the risk of death from prostate cancer when accounting for competing risk of death.Material and methodsWe investigated a nationwide cohort of 15,878 prostate cancer patients diagnosed in 2010–2014; with 3343 deaths occurring through 2016. Blinded medical chart review was carried out for 670 deaths and compared to the national cause of death registry. Five death categories were defined: 1) prostate cancer-specific death, 2) other unspecified urological cancer death, 3) other cancer death 4) cardiovascular disease death, and 5) other causes of death. Competing risk analyses compared Cox cause-specific and Fine-Gray regression models.ResultsChart review attributed 51.2% of deaths to prostate cancer, 17.0% to cardiovascular disease, and 16.7% to other causes. The Danish Register of Causes of Death attributed 71.7% of deaths to prostate cancer when including all registered contributing causes of death, and 57.0% of deaths when including only the primary registered cause of death. The probability of death by prostate cancer was 10% at 2-year survival.ConclusionsMore than half of the deceased men in our study cohort died of their prostate cancer disease within a mean of 2.4 years of follow up. Data from the death registry is prone to misclassification, potentially overestimating the proportion of deaths from prostate cancer.