一氧化氮及相关物质在卵巢肿瘤患者外周血中的检测

目的探讨一氧化氮(NitricOxideNO)及其相关物质谷胱甘肽(GlutathioneGSH)的含量及超氧化物歧化酶(SuperoxidedismutaseSOD)活性的变化意义.方法应用改良的镀铜镉还原法测定NO2-/NO3-的含量,二硫双苯甲酸(DTNB)法测定GSH的含量,黄嘌呤氧化酶法测定SOD的活性.取卵巢肿瘤患者的外周血共74例,其中良性肿瘤40例,恶性肿瘤34例.另外取经体检健康女性的外周血共70例.将以上三组标本所测的数据用方差分析作比较.每组标本中三项指标进行相关分析.结果在正常人与卵巢良性肿瘤患者外周血的比较中,NO、GSH、SOD均无显著差异;在正常人与卵巢恶性肿瘤患者的外周血的比较中,NO及GSH的含量和SOD的活性均有显著差异(P＜0.05),正常人显著高于恶性肿瘤患者.在卵巢良性肿瘤患者与恶性肿瘤患者的比较中,NO及GSH的含量均有显著差异(P＜0.05),良性肿瘤患者显著高于恶性肿瘤患者.二者之间SOD的活性有显著差异(P＜0.05),良性肿瘤患者显著高于恶性肿瘤患者.正常人的外周血中NO与GSH显著正相关,r＝0.672(P＜0.01);NO与SOD,GSH与SOD均无相关关系.在卵巢良、恶性肿瘤患者的外周血中NO、GSH与SOD三者之间无相关关系.结论NO本身及其相关物质GSH,SOD含量降低可能与卵巢恶性肿瘤有一定关系.     

一氧化氮及相关物质在卵巢肿瘤患者外周血中的检测

目的：探讨一氧化氮（NitricOxideNO）及其相关物质谷胱甘肽（GlutathioneGSH）的含量及超氧化物歧化酶（Superoxide dismutaseSOD）活性的变化意义。方法：应用改良的镀铜镉还原法测定NO2^-/NO3^-的含量，二硫双苯甲酸（DTNB）法测定GSH的含量，黄嘌呤氧化酶法测定SOD的活性。取卵巢肿瘤患者的外周血共74例，其中良性肿瘤40例，恶性肿瘤34例。另外取经体检健康女性的外周血共70例。将以上三组标本所测的数据用方差分析作比较。每组标本中三项指标进行相关分析。结果：在正常人与卵巢良性肿瘤患者外周血的比较中，NO、GSH、SOD均无显著差异；在正常人与卵巢恶性肿瘤者的外周血的比较，NO及GSH的含量和SOD的活性均有显著差异（P＜0．05），正常人显著高于恶性肿瘤患者。在卵巢良性肿瘤患者与恶性肿瘤患者的比较中，NO及GSH珠含量均有显著差异（P＜0．05），良性肿瘤患者显著高于恶性肿瘤患者。二者之间SOD的活性有显著差异（P＜0．05），良性肿瘤患者显著高于恶性肿瘤患者、正常人的外周血中NO与GSH显著正相关，r＝0．672（P＜0．01）；NO与SOD，GSH与SOD均无相关关系，在卵巢良、恶性肿瘤患者的外周血中NO、GSH与SOD三者之间无相关关系。结论：NO本身及其相关物质GSH，SOD含量降低可能与卵巢恶性肿瘤有一定关系。     

骨肉瘤及其周围肌肉组织中NOS和NO的测定及其意义

目的　研究骨肉瘤及其周围肌肉组织中一氧化氮合酶 (NOS)活性与一氧化氮 (NO )含量之间的关系。方法　采用分光光度法对 37例骨肉瘤组织 ,分别测定其中NOS活性和NO含量 ,同时分别测定肿瘤周围正常肌肉组织中NOS活性和NO含量 ,然后进行相应的统计学分析。结果　肿瘤组织和肿瘤周围正常肌肉组织中NOS活性分别为 ( 0 .15± 0 .0 5 )U /mg蛋白质和( 1.88± 0 .2 7)U /mg蛋白质 ,两者比较有非常显著性差异 (P <0 .0 1) ,NO含量平均分别为 ( 46.43± 2 5 .98) μmol/L和 ( 49.0 8±1.5 0 ) μmol/L ,两者差异不明显 ;转移性瘤组织与非转移性瘤组织中NOS活性分别为 ( 0 .16± 0 .0 4)U /mg蛋白质和 ( 0 .13±0 .0 6)U /mg蛋白质 ,两者差异不明显 ,NO含量分别为 ( 36.44± 2 7.2 1) μmol/L和( 5 6.43± 2 0 .82 ) μmol/L ,两者比较有显著性差异 (P <0 .0 5 ) ;转移性瘤周围正常肌肉组织与非转移性瘤周围正常肌肉组织中的NOS活性分别为 ( 1.2 3± 0 .34 )U /mg蛋白质和 ( 2 .5 4±0 .5 5 )U /mg蛋白质 ,两者比较有显著性差异 (P <0 .0 5 ) ,NO含量分别为 ( 40 .72±1.5 4) μmol/L和 ( 5 7.46± 2 .18) μmol/L ,两者比较有显著性差异 (P <0 .0 5 )。结论　骨肉瘤组织中的NO是由肿瘤组织及肿瘤周围组织共同产生     

口腔鳞癌患者血清一氧化氮水平及其手术前后的变化

目的 :探讨口腔鳞癌患者血清一氧化氮水平及手术前后的变化和意义。方法 :采用化学比色法对 41例口腔鳞癌、31例口腔颈面部良性肿瘤及 2 0例健康人血清一氧化氮 (NO)和一氧化氮合成酶 (NOS)水平进行了检测。结果 :口腔鳞癌组血清NO和NOS水平显著高于良性病变组和健康对照组 (P <0 .0 5) ,口腔鳞癌组中NO和NOS水平在有颈淋巴结转移者显著高于无颈淋巴结转移者 (P <0 .0 5) ,病变切除手术后NO及NOS显著低于手术前 (P <0 .0 5)。结论 :NO和NOS与口腔鳞癌的发生、发展有密切关系 ,NO和NOS的检测对口腔鳞癌的病情判定和指导治疗有重要的意义     

非小细胞肺癌患者血清及肿瘤组织一氧化氮合酶活性变化及临床意义

目的　研究非小细胞肺癌患者血清和肿瘤组织中一氧化氮合酶 (NOS)的活性变化及其临床意义。方法　采用分光光度法检测 30例非小细胞肺癌患者血清及肿瘤组织中 NOS的活性 ,并分析其临床意义。结果　 (1)肺癌患者血清 NOS活性较正常人显著增高 (P<0 .0 1) ;(2 )术后血清 NOS活性较术前显著降低 (P<0 .0 1) ;(3)血清及肿瘤组织中 NOS活性随肺癌的恶性程度加重而增高 (P<0 .0 1)。结论　非小细胞肺癌患者血清及肿瘤组织中NOS活性的增高与肿瘤的生长及恶性行为有关     

TRAIL促人甲状腺癌细胞凋亡中一氧化氮作用的探讨

目的:探讨一氧化氮(NO)在肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导人甲状腺癌细胞凋亡中的作用。方法:S-亚硝基化生物素转化法、一氧化氮合酶(NOS)活性及NO产物测定法检测各组细胞3-磷酸甘油醛脱氢酶(GAPDH)S-亚硝基化与NO产物关系;蛋白质印迹法检测各组细胞及细胞核中GAPDH蛋白表达;台盼蓝染色、DNA裂解分析、Caspase-3活性测定法检测各组FRO细胞凋亡。结果:20ng/mLTRAIL处理FRO细胞0～24h,可见NOS活性及作为NO氧化产物的硝酸盐和亚硝酸盐水平呈时间依赖性增加,于4h开始显著增高,P=0.008;12h达高峰。iNOS抑制剂L-NAME抑制TRAIL诱导的GAPDHS-亚硝基化及其随后的细胞核转位,但不影响TRAIL诱导的GAPDH表达。L-NAME显著抑制TRAIL诱导的人甲状腺癌细胞凋亡和Caspase-3活性,P值均<0.001。结论:NO介导的GAPDHS-亚硝基化修饰和随后的细胞核转位可能参与了TRAIL诱导的人甲状腺癌细胞凋亡。     

阿霉素对兔心功能及心肌一氧化氮合酶表达的影响

目的：探讨阿霉素对兔心功能及心肌一氧化氮合酶（NOS）表达的影响。方法：测定实验组和对照组兔心输出量（CO）、左心室收缩压（LVSP）、左心室舒张末压（LVEDP）。取心肌组织进行HE染色和NADPH组织化学染色并进行图像定量分析,所测得数据均进行统计学处理。结果：实验组CO、LVSP均明显低于对照组（P＜0．01）,LVEDP明显高于对照组（P＜0．01）,心肌组织NADPH组织化学染色及图像定量分析结果显示,实验组NOS表达明显增强（P＜0．01）,且与CO呈负相关（γ=-0．8304,P＜0．01）,与LVSP呈负相关（γ=0．7815,P＜0．05）,与LVEDP呈正相关（γ＝0．7356,P＜0．05）。结论NOS过度表达,导致NO生成过量,参与阿霉素慢性心功能损伤的过程。     

一氧化氮合酶mRNA和蛋白在骨巨细胞瘤中的表达及意义

背景与目的：近年的研究发现,一氧化氮在恶性肿瘤的发生和发展中起着重要作用,且在正常骨细胞中检测到一氧化氮合酶(nitric oxide synthase,NOS)的活性,但目前未见有关骨巨细胞瘤(giant cell tumors of bone,GCT)NOS的活性及其与GCT病理分级和肿瘤复发关系的报道。本研究探讨GCT组织中NOS mRNA和蛋白的表达,及其与GCT病理分级、肿瘤复发的关系。方法：用NOS的cDNA探针检测14例冻存GCT组织中原生型一氧化氮合酶(constitutive NOS,cNOS)、诱导型一氧化氮合酶(inducible NOS,iNOS)mRNA的表达,用抗NOS1、NOS2、NOS3的多克隆抗体检测42例GCT石蜡包埋组织中NOSl、NOS2、NOS3蛋白的表达。结果：(1)在14例冻存的GCT组织中多核巨细胞(multinuclear giant cell,MGC)cNOS和iNOS mRNA的阳性率分别为78．6％和57．1％,单核细胞(mononuclear cell,MC)的阳性率均为35．7％;(2)病理分级Ⅱ、Ⅲ级MGC的cNOS mRNA阳性率明显高于I级MGC(P=0．008);(3)在42例GCT的石蜡包埋组织中,MGC中NOS1、NOS2、NOS3蛋白的阳性率分别为85．7％、59．5％和31．0％,MC的阳性率分别为54．8％、28．6％和14．3％;(4)病理分级Ⅱ、Ⅲ级GCT组织中MC的NOS1阳性率明显高于I级组(P=0．006);(5)复发组GCT组织MC的NOS1阳性率明显高于无复发组(P=0．018),复发组GCT组织中MGC的NOS3蛋白阳性率明显高于无复发组(P=0．041)。结论：GCT组织中NOS的表达,尤其是cNOS在MC中的表达,与病理分级和肿瘤复发有密切关系。     

一氧化氮合酶与浸润性乳腺癌血管形成的关系及临床意义

目的:探讨一氧化氮合酶(nitrioxide synthase,NOS)与浸润性乳腺癌新生血管形成的关系及其临床意义。方法:采用分光光度法检测30例浸润性乳腺癌及其相应癌旁乳腺组织和30例良性乳腺病变组织的 NOS活性,应用免疫组织化学SP法,以抗第8因子相关抗原(ⅧRAg)抗体标记血管内皮细胞,根据ⅧRAg阳性的血管内皮细胞计数来测定微血管密度(microvessel density, MVD)。结果:1)浸润性乳腺癌 NOS活性(2. 22±1 .39)显著高于相应癌旁乳腺组织(1. 29±0 .48)及良性乳腺病变组织(0 .50±0. 14),P=0 .004,P=0. 000;相应癌旁乳腺组织NOS活性高于良性乳腺病变组织,P=0 .000。2)乳腺癌 MVD计数(40. 53±8. 29)明显高于相应癌旁乳腺组织(18. 07±4. 90),P=0 000。3)有腋淋巴结转移者 NOS活性(2 .60±1 .66)高于无淋巴结转移者(1 .72±0 .41),P=0 .048,且明显形成更多的微血管,P=0 000。4)乳腺肿瘤组织NOS活性和MVD与肿瘤大小、分化程度和ER、PR表达状况无关。5)浸润性乳腺癌NOS活性与肿瘤 MVD呈正相关,r=0 .494,P=0 .014。结论:1)浸润性乳腺癌NOS活性、MVD与其淋巴结转移密切相关,一氧化氮(nitric oxide, NO)生成可促肿瘤新生血管形成,并促其生长、发展和侵袭转移。2)肿瘤微环境中 NO合成增加可能促肿瘤生长。3)NOS活性增加可作     

肺癌与一氧化氮合酶表达关系的研究

目的研究人体肺癌组织中一氧化氮合酶各亚型的表达及其与肺癌生物学行为的关系.方法采用免疫组织化学方法检测24例人体肺癌组织中一氧化氮合酶三种亚型的表达情况,并采用计算机自动图像分析系统对免疫组化染色强度进行测定.结果肺癌组织中有三种亚型的一氧化氮合酶表达,以NOS1和NOS3表达为主,其中,NOS1在腺癌中的表达强于鳞癌(P＜0.05),NOS1和NOS3在临床一、二期肺癌的表达高于三、四期肺癌(P＜0.001),无淋巴结转移者NOS1和NOS3的表达高于淋巴结转移者(P＜0.02和P＜0.001).结论NOS尤其是NOS1和NOS3在肺癌组织中的表达可以抑制肺癌的进展.     

5—氟脲嘧啶对恶性肿瘤患者血一氧化氮水平的影响

目的 探讨５－氟脲嘧啶（５－Ｆｕ）对恶性肿瘤患者血一氧化氮（ＮＯ）水平的影响。方法 测定５２例恶性肿瘤患者５－Ｆｕ化疗前后血中一氧化氮合成酶（ＮＯＳ）和ＮＯ的水平,并进行统计学分析。结果 肿瘤患者的ＮＯＳ、ＮＯ水平明显高于健康人（Ｐ〈０．０１）,而未手术切除者又明显高于手术切除者（Ｐ〈０．０５和Ｐ〈０．０１）;５－Ｆｕ化疗后ＮＯＳ、ＮＯ水平明显降低（Ｐ〈０．０１）,与健康人无显著性差异（Ｐ〉０．０     

胃癌组织和血清一氧化氮及一氧化氮合成酶定量分析

目的：探讨一氧化氮（ＮＯ）与胃癌的关系。方法：应用比色法测定了３１例胃癌患者血清和组织的ＮＯ及一氧化氮合成酶（ＮＯＳ）含量，并设２５例正常人和２６例胃十二批肠溃疡为血清对照组。结果：胃癌患者血清ＮＯ水平明显低于正常对照组及胃十二指肠溃疡组（Ｐ〈０．０１）；而溃疡组则明显高于正常对照组（Ｐ〈０．０５）；胃癌组织中ＮＯ及ＮＯＳ含量则高于癌旁组织（Ｐ〈０．０５）。结论：当胃癌发生时，巨噬细胞产大大量的Ｎ     

外源一氧化氮对胃癌SGC-7901细胞增殖及细胞周期的影响

目的:探讨外源一氧化氮(nitric oxide, NO)对胃癌细胞生长和增殖的影响.方法:应用MTT法评价NO供体硝普钠(sodium nitroprusside,SNP)和一氧化氮合酶(nitric oxide synthase,NOS)抑制剂N-硝基-L-精氨酸甲酯(N-nitro-L-arginine methylester,L-NAME)对胃癌SGC-7901细胞生长的抑制作用,RT-PCR和Western印迹法检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)和Caspase-3 mRNA和蛋白的表达,FCM法检测细胞周期的变化.结果:SNP使细胞周期发生变化,与对照组比较,S期细胞减少,G_0/G_1期细胞增多,细胞由IC50/G_1期进入S期延缓;SNP抑制SGC-7901细胞的生长,PCNA mRNA、PCNA蛋白和Caspase-3蛋白表达降低.而NOS抑制剂L-NAME预处理逆转了SNP的这种作用.结论:NO可抑制胃癌细胞生长和增殖,加速细胞凋亡.     

Nitric oxide synthase and vascular endothelial growth factor expression in hepatocellular carcinoma and the correlation with angiogenesis

It is now well established that tumor growth and metastasis are angiogenesis dependent[1]. Since Weidner reported microvessel density (MVD) was useful in prognosis of patients with breast cancer[2], the prognostic value of MVD have been certified in several tumors including primary liver cancer[3]. The increasing activity of Nitric oxide synthase (NOS) correlated with tumor angiogenesis[4]. We examined HCC tissue sections immunohistochemically for expression of iNOS, eNOS, VEGF, and …     

Nitric Oxide Synthase Activity in Human Lung Cancer

Nitric oxide synthases (NOS) exist in human tumor cell lines and solid tumor tissues, and it has been suggested that NO may play important roles in growth, progression or metastasis of tumors. We investigated the activity and distribution of NOS in a series of human lung cancer and normal lung tissues. Seventy-two primary lung cancer samples (44 cases of adenocarcinoma, 18 of squamous cell carcinoma, 4 of large cell carcinoma, 2 of small cell carcinoma, 2 of adenosquamous carcinoma, and 2 of carcinoids) and corresponding normal lung samples were obtained from surgically treated patients. In normal lung tissues, little NOS activity was observed with no correlation between the patients' age and NOS activity. The total NOS activities in lung adenocarcinoma samples were significantly higher than those in other types of lung cancers or normal lung samples (P<0.05), Analysis by tumor grade of the adenocarcinoma samples revealed no significant difference of NOS activity between grades. TNM classification showed that, although T stage did not correlate with NOS activity, cancer tissues from patients with N2 disease tended to have lower activity than those from patients with NO or Nl disease. Immunohistochemical studies revealed that the intensity of NOS immunoreactivity correlated with NOS activity. These results suggest that NO may play an important role in the metabolism and behavior of lung cancers, especially adenocarcinoma.     

一氧化氮/一氧化氮合酶信号系统与前列腺癌

一氧化氮(NO)作为体内重要的信使分子,其生物学作用复杂、多样,与一氧化氮合酶(NOS)活性及表达密切相关.NO通过G蛋白受体激活的信号转导途径参与肿瘤细胞增殖、迁移和血管新生,与前列腺癌的发生、发展密切相关.NO供体药物及NOS抑制剂具有抗肿瘤和放化疗增敏的作用.     

肝细胞癌中NOS和VEGF的表达及其与肿瘤血管形成的关系

目的 研究Ⅱ、Ⅲ型一氧化氮合酶（ｉＮＯＳ、ｃＮＯＳ）和血管内皮生长因子（ＶＥＧＦ）在肝细胞癌中的表达及其与肿瘤血管形成的关系。方法 应用免疫组化方法检测７１例肝细胞癌患者手术切除石蜡包埋标本ｉＮＯＳ、ｅＮＯＳ、ＶＥＧＦ的表达,抗ＣＤ３４单克隆抗体显示血管内皮细胞。根据ＣＤ３４阳性的血管内皮细胞计数来测定肿瘤微血管密度（ＭＶＤ）。结果 ｉＮＩＳ、ｅＮＯＳ分别在８１．３％、８５．９％的肝癌中阳性表达,     

Arginase 2 is expressed by human lung cancer, but it neither induces immune suppression, nor affects disease progression

In human prostate cancer, Arginase 2 (ARG2) and nitric oxide synthase (NOS) are concomitantly expressed by tumor cells, and induce tumor immune escape via peroxynitrite-dependent Tyrosine nitrosylation. Since there were no data regarding this immune suppressive mechanism in other tumor types, and an evaluation of its clinical relevance in human tumors had still to be provided, we have investigated presence and clinical relevance of ARG2 and NOS expression in lung cancer. No evidence of NOS expression was found, no significant NOS enzymatic activity was detected. Instead, ARG2 protein was expressed by tumor cells. In a cohort of 120 patients, the amount of ARG2-positive tumor cells was significantly higher in small cell lung cancers (SCLC) than in non-small cell lung cancers (NSCLC). Large cell undifferentiated carcinomas had twice ARG2 than the other NSCLC subtypes. ARG2 expression was increased in Grade 3 tumors, as compared to Grades 1 and 2. However, no relationship was found with tumor size and stage, and with patient survival. Indeed, the enzyme was active, since the Arginine catabolite Ornithine was produced, but Arginine depletion was not attained. In addition, nitrotyrosine was not found in tumor tissue. Accordingly, when tumor cells isolated from lung cancer were incubated with activated autologous T cells, no inhibition of proliferation was detected. Our results indicate that ARG2 is expressed in lung cancer, but it does not induce tumor immune escape and does not affect disease progression, most probably due to the lack of concomitant NOS expression.     

Pro-survival and pro-growth effects of stress-induced nitric oxide in a prostate cancer photodynamic therapy model

We discovered recently that human breast cancer cells subjected to photodynamic therapy (PDT)-like oxidative stress localized in mitochondria rapidly upregulated nitric oxide synthase-2 (NOS2) and nitric oxide (NO), which increased resistance to apoptotic photokilling. In this study, we asked whether human prostate cancer PC-3 cells would exploit NOS2/NO similarly and, if so, how proliferation of surviving cells might be affected. Irradiation of photosensitized PC-3 cells resulted in a rapid (<1 h), robust (∼12-fold), and prolonged (∼20 h) post-irradiation upregulation of NOS2. Caspase-3/7 activation and apoptosis were stimulated by NOS2 inhibitors and a NO scavenger, implying that induced NO was acting cytoprotectively. Cyclic GMP involvement was ruled out, whereas suppression of pro-apoptotic JNK and p38 MAPK activation was clearly implicated. Cells surviving photostress grew back ∼2-times faster than controls. NOS2 inhibition prevented this and the large increase in cell cycle S-phase occupancy observed after irradiation. Thus, photostress upregulation of NOS/NO elicited both a pro-survival and pro-growth response, both of which could compromise clinical PDT efficacy unless suppressed, e.g. by pharmacological intervention with a NOS2 inhibitor.     

一氧化氮在肿瘤性贫血患者中的表达

目的研究肿瘤性贫血患者血清一氧化氮(NO)和一氧化氮合酶(NOS)的表达水平,探讨其与肿瘤性贫血的关系。方法选择2009年9月至2011年10月期间的肿瘤性贫血患者40例,为观察组;另外选择同期体检的健康人15例,为对照组。采用硝酸还原酶法测定NO和NOS含量。结果肿瘤性贫血患者NO和NOS水平明显高于正常对照(P<0.05);随着贫血程度加重,NO和NOS水平明显升高(P<0.05)。结论 NO在肿瘤性贫血的发病中起重要作用,及早降低NO水平,有助于阻止贫血的发展,可以为肿瘤性贫血的治疗提供新策略。