诱导型一氧化氮合酶与PTEN在鼻咽癌中的表达及意义

诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和第10号染色体缺失的与张力蛋白同源的磷酸酯酶基因(phospharase and tensill homolog deleted on chromosome ten, PTEN)的联合表达与一些肿瘤的生物学行为有关~([1-2]).     

一氧化氮、诱生型一氧化氮合酶和血管内皮生长因子C在鼻咽癌中的作用

目的检测鼻咽癌组织中诱生型一氧化氮合酶（inducible nitric oxide synthase,iNOS）及血管内皮生长因子C（vascular endothelial growth factor C,VEGF-C）表达的意义以及它们之间的关系,并检测一氧化氮（nitric oxide,NO）对鼻咽癌细胞株生长的影响及对VEGF-C表达的调节作用。方法用免疫组化法检测57例鼻咽癌标本以及20例非鼻咽癌鼻咽上皮组织标本中VEGF-C以及iNOS的表达,另以NO供体S-亚硝基-N-乙酰基-DL-青霉胺（S-nitroso-N-acetylpenicillamine,SNAP）对鼻咽癌细胞系TWO3进行处理,通过MTT法检测NO对TWO3生长的影响,并通过逆转录聚合酶链反应（RT-PCR）法检测NO对TWO3的VEGF-C表达调控作用。结果VEGF-C和iNOS在鼻咽癌组中表达阳性率显著高于对照组（P〈0.05）,鼻咽癌组织中VEGF-C表达在不同T、N以及临床分期组差异有统计学意义（P〈0.05）,但是不同年龄性别组表达差异无统计学意义（P〉0.05）。鼻咽癌组织中iNOS的表达在不同T、N级,临床分期,年龄以及性别组中表达差异无统计学意义（P〉0.05）;鼻咽癌组织中iNOS与VEGF-C表达显著相关（P〈0.05）;高浓度NO能抑制TWO3生长,且NO能调节TWO3的VEGF-C的转录表达。结论VEGF-C及iNOS可能对鼻咽癌的发展起促进作用,且在鼻咽癌中NO可能调节VEGF以及VEGF-C表达调节。     

诱导型一氧化氮合酶mRNA在分泌性中耳炎中表达的意义

目的探讨分泌性中耳炎(secretory ototis media，SOM)患者中耳积液及外周血白细胞诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)mRNA表达及在SOM发病过程中的意义，以及与细菌感染和免疫介导的关系。方法随机取45例SOM患者及30例健康人的外周血白细胞，并抽取SOM患者患耳中耳积液，用原位杂交方法检测iNOS—mRNA。结果健康人外周血白细胞未见iNOS—mRNA表达。SOM患者外周血白细胞iNOS—mRNA表达：急性组阳性率为63．64％，镜下阳性细胞率为60．3％；亚急性组阳性率为8．7％，镜下阳性细胞率为72．5％；SOM患者中耳积液iNOS—mRNA表达：急性组阳性率为45．45％，镜下阳性细胞率为80％；亚急性组阳性率为52．17％，镜下阳性细胞率为84％。SOM患者外周血白细胞及中耳积液中iNOS—mRNA表达高度增强，其中在急性组外周血白细胞中表达显著高于亚急性组，而中耳积液iNOS—mRNA表达阳性率及阳性细胞率在急性组与亚急性组中表达强度无显著性差异。结论诱导型一氧化氮合酶(iNOS)及诱导型一氧化氮合酶——氧化氮(iNOS—NO)通路在SOM的发病、中耳积液的形成过程中可能起着重要作用。     

诱导型一氧化氮合酶在变应性真菌性鼻-鼻窦炎黏膜中的表达

目的 :通过对变应性真菌性鼻 鼻窦炎窦腔病变黏膜 (AFRS)和慢性鼻窦炎 (CRS)病变黏膜中诱导型一氧化氮合酶 (iNOS)的定位和半定量研究 ,探讨AFRS和CRS发病机制的差异 ,一氧化氮 (NO)的生成情况和意义。方法 :用免疫组化 (S P法 )的方法对 2 8例AFRS和 6例CRS标本中iNOS进行定位和半定量检测。并且用Ridit分析进行统计分析。结果 :iNOS广泛存在于上皮组织、血管内皮、平滑肌细胞、黏膜下浆液性腺体和炎症细胞中 ,以细颗粒或粗颗粒存在于细胞质中 ,细胞膜上偶见。细胞核及间质细胞内无iNOS表达。AFRS黏膜与CRS黏膜之间的表达差异有显著性意义 (P <0 .0 1)。结论 :iNOS在AFRS窦腔黏膜中呈大量表达 ,iNOS对AFRS发病起重要作用 ;AFRS与CRS可能是不同的两个疾病     

变应性鼻炎患者外周血白细胞及鼻黏膜诱导型一氧化氮合酶mRNA表达的意义

OBJECTIVE: To investigate the relationship between expression of leukocyte inducible nitric oxide synthase (iNOS)-mRNA and allergic rhinitis (AR). METHOD: Thirty-five patients with AR and 30 healthy controls were included in this study. Expression of iNOS-mRNA in peripheral blood leukocyte was detected by in situ hybridization. NO in plasm was measured by nitrate reductase. Expression of iNOS-mRNA in nasal mucosal was detected in 8 patients with AR and 6 healthy controls. RESULT: No expression of leukocyte iNOS-mRNA in healthy controls was found. In AR patients, the positive cells were significantly increased, the positive rate reached 40.82%. Expression of iNOS-mRNA was localized at the epithelium, gland and macrophage in healthy controls. Hyperplasia and expression of iNOS-mRNA increased at epithelium, gland and macrophage in the AR patients(t = 23.17, P < 0.001). The level of plasm NO in AR group was higher than that in healthy control group (t = 27.89, P < 0.001). CONCLUSION: There is a relationship between expression of leukocyte iNOS-mRNA and the level of plasm NO in AR patients. The study provides an easy method of in situ hybridization for detecting some signal in body.     

缺氧诱导因子-1α及诱导型一氧化氮合酶在中耳胆脂瘤上皮中的表达及意义

目的：研究缺氧诱导因子-1α（HIF-1α）、诱导型一氧化氮合酶（iNOS）在中耳胆脂瘤上皮中的表达及意义,探讨其在发病机制中的可能作用。方法：运用免疫组织化学SP法检测21例胆脂瘤标本及11例正常外耳道皮肤标本中HIF-1α、iNOS的表达。结果：在21例中耳胆脂瘤上皮及11例正常外耳道皮肤中,HIF-1α的表达指数分别为52．49±13．80、0．60±0．49,两者比较差异有统计学意义（P〈O．01）;iNOS的表达指数分别为92．05±27．84、1．15±0．84,两者比较差异有统计学意义（P〈0．01）;HIF-1α与iNOS的表达指数呈正相关（r=0．536,P〈0．05）。结论：HIF-1α及iNOS蛋白在中耳胆脂瘤上皮中存在高度表达;缺氧及其相关因子HIF-1α、iNOS等可能在中耳胆脂瘤的发生、发展过程中起着重要促进作用。     

变应性鼻炎患者外周血白细胞及鼻黏膜诱导型一氧化氮合酶mRNA 表达的意义

目的　探讨变应性鼻炎 (allergicrhinitis,AR)患者外周血白细胞及鼻黏膜诱导型一氧化氮合酶 (induciblenitricoxidesynthase,iNOS)mRNA表达的关系。方法　选择 3 5例AR患者及 3 0例健康人的外周血白细胞。其中 8例变应性鼻炎患者鼻黏膜 ,6例正常鼻黏膜。iNOS mRNA表达采用原位杂交方法。血浆一氧化氮 (nitricoxide,NO)水平采用硝酸还原酶比色法测定。结果　健康人外周血白细胞未见iNOS mRNA表达 ,而AR患者外周血白细胞iNOS mRNA表达高度增强 ,其阳性率达 40 82 %。正常人鼻黏膜上皮、腺体及巨噬细胞可见iNOS mRNA的低度表达 ,而AR患者中上皮、腺体及巨噬细胞增生 ,并iNOS mRNA表达高度增强 (t=2 3 17,P <0 0 0 1)。AR组血浆NO水平显著高于对照组 (t=2 7 89,P <0 0 1)。结论　AR患者血浆NO水平与外周血白细胞及组织内iNOS mRNA表达高度增强有关。提示iNOS NO通路在AR的发病过程中可能起重要作用。本研究为检测体内某些信号提供了简便易行的原位杂交试验方法     

The effect of corticosteroid therapy on cyclooxygenase 2, vascular endothelial growth factor,and inducible nitric oxide synthase expression levels in nasal polyposis

Nasal and oral corticosteroid therapy is the ultimate treatment for sinonasal polyposis. Although there are numerous clinical studies regarding the factors associated with the formation of nasal polyposis, there is not enough literature on how these factors are influenced by steroid treatment. Twenty-one patients that had no prior medical therapy for nasal polyposis or had received medical therapy at least 6 months earlier were included in the study. Patients were treated with oral and nasal corticosteroid therapy. Nasal polyp biopsies were taken before and after medical treatment and immunohistochemical staining for cyclooxygenase 2 (COX-2), vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) were applied to the specimens. In this study, we tried to demonstrate the effects of corticosteroid therapy on nasal polyposis tissue immunohistochemically. There was no change at immunohistochemical expression level of COX-2; however, the decline of immunohistochemical expression levels of VEGF and iNOS was statistically significant. Short-term steroid therapy does not affect COX-2 level of the nasal polyposis tissue, but has an influence on iNOS and VEGF levels. Our findings were harmonious with those of the previous studies of the literature. Further studies are needed to demonstrate the long-term effects with a larger patient group.     

喉癌喉咽癌组织中诱导型一氧化氮合酶的表达及临床意义

目的 :探讨诱导型一氧化氮合酶 (iNOS)在喉癌喉咽癌组织中的表达及其临床意义。方法 :采用逆转录 聚合酶链反应 (RT PCR)技术检测 4 0例喉癌及 12例喉咽癌 (喉癌喉咽癌组 )、5 0例癌旁组织 (癌旁组 )、12例良性肿瘤 (良性肿瘤组 )及 7例正常喉黏膜 (正常喉黏膜组 )组织中iNOSmRNA的表达。结果 :喉癌喉咽癌组阳性表达率为 78.85 % ,与正常喉黏膜组 (0 % )和癌旁组 (2 0 .0 0 % )比较 ,差异有极显著性意义 (P <0 .0 1) ;与良性肿瘤组 (4 1.6 7% )比较 ,差异亦有显著性意义 (P <0 .0 5 )。有局部淋巴结转移组 (95 .2 4 % )显著高于无局部淋巴结转移组 (6 7.74 % ) (P <0 .0 5 ) ;iNOSmRNA阳性表达与T分级呈正相关 (P <0 .0 5 ) ,与细胞分化程度呈负相关 (P <0 .0 5 )。结论 :喉癌喉咽癌组织中iNOSmRNA呈高表达 ,提示iNOS可能通过合成NO在分子水平参与了喉癌喉咽癌的发生、发展 ;NO供体药物和iNOS特异性抑制剂的开发 ,必将为肿瘤治疗带来可喜的前景。     

Association of vascular endothelial growth factor expression with angiogenesis and lymph node metastasis in nasopharyngeal carcinoma.

OBJECTIVE: Recent experimental evidence indicates that angiogenesis affects tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is considered to be an important regulator of tumor angiogenesis. The present study was designed to examine the role of VEGF on angiogenesis and lymph node metastasis in primary nasopharyngeal carcinomas (NPCs). STUDY DESIGN: Formalin-fixed paraffin-embedded biopsy specimens were obtained from 29 primary NPCs that consisted of 22 differentiated nonkeratinizing carcinomas and seven undifferentiated carcinomas. METHODS: Microvessels were highlighted by staining endothelial cells with von Willebrand factor (VWF) using immunohistochemical techniques, and were counted (per x 400 field) in the most active area of angiogenesis on light microscopy. The expression of VEGF was also studied with immunohistochemistry. Positive ratio for VEGF was graded on a scale of 1 and 2. Scale 1 represents patients with less than the mean value of the positive ratio, and scale 2 represents patients with more than the corresponding value. RESULTS: There was a significant correlation between increased microvessel count and the progression of regional lymph node involvement. The microvessel counts and the progression of N factor were significantly higher in scale 2 patients than in scale 1 patients. CONCLUSION: These results suggest that VEGF plays an important role in lymph node metastasis through induction of angiogenesis in NPCs.     

血管内皮生长因子在鼻咽癌组织中的表达及其与淋巴转移的关系

目的 检测鼻咽癌中血管内皮生成因子C(VEGF-C)的表达及微淋巴管密度(microlymphatic vessel density,MLVD),探讨鼻咽癌淋巴管生成及淋巴转移之间的关系.方法 采用免疫组化SP法检测58例鼻咽癌和20例鼻咽部炎性反应组织中VEGF-C的表达情况,并用淋巴管内皮细胞特异性抗体LYVE-1行免疫组化染色,计数肿瘤内MLVD,并结合临床病理特征进行分析.结果 鼻咽癌组和炎性反应对照组VEGF-C的阳性表达率分别为84.5%(49/58)和15.0%(3/20),差异有统计学意义(X2=32.309,P＜0.01).鼻咽癌组织MLVD为(28.6±6.2)个/视野,鼻咽炎性反应组为(10.5 4±3.0)个/视野,两组差异有统计学意义(t=12.491,P＜0.01).鼻咽癌组织中,有淋巴转移组VEGF-C阳性表达率(87.8%)明显高于无淋巴转移组(76.5%);有淋巴转移组MLVD(30.2 4±6.4)个/视野高于无淋巴转移组(24.8±3.6)个/视野,经统计学分析,差异均有统计学意义(t=3.259,P＜0.01).VEGF-C的表达与MLVD(t=3.512,P＜0.01)、淋巴转移(X2=7.715,P＜0.01)、临床分期(X2=4.250,P＜0.05),与病理分化程度无关(X2=0.000,P＞0.05).VEGF-C的表达与MLVD(t=3.512,P＜0.01)、淋巴转移(X2=7.715,P＜0.01,r=0.712)、临床分期(X2=4.250,P＜0.05,r=0.481)相关,与病理分化程度无关(X2=0.000,P＞0.05).结论 在鼻咽癌组织中VEGF-C呈高表达,VEGF-C表达与鼻咽癌组织MLVD、淋巴转移、临床分期密切相关.VEGF-C可能参与了鼻咽癌发生、浸润和转移的过程.VEGF-C与肿瘤组织微淋巴管密切相关,在鼻咽癌发展过程中起重要作用,可能成为抗肿瘤治疗的潜在靶点.     

鼻咽癌患者血清血管内皮生长因子检测及临床意义

目的 :探讨鼻咽癌患者放疗前后血清中血管内皮生长因子 (VEGF)的变化及临床意义。方法 :应用酶联免疫吸附法 (ELISA)对 4 6例鼻咽癌患者放疗前后血清中VEGF水平进行检测。结果 :鼻咽癌患者放疗前血清VEGF水平明显高于正常人 (P <0 .0 1) ,并随病情进展而升高 ;放疗后VEGF水平较放疗前明显降低 (P <0 .0 1)。VEGF水平 >15 0ng/L与 <15 0ng/L者的复发或转移率有明显差异 (P <0 .0 1)。结论 :VEGF不仅与鼻咽癌发生、发展有关 ,而且与其预后也密切相关 ,可望作为鼻咽癌患者预后的一个新的检测指标     

HIF-1α、VEGF在阻塞性睡眠呼吸暂停低通气综合征患者

目的 通过研究阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者软腭组织中HIF-1α、VEGF的表达,探讨HIF-1α、VEGF表达与OSAHS的关系及意义。 方法 选取 30 例 OSAHS 患者作为实验组, 6 例排除OSAHS 的单纯慢性扁桃体炎患者作为对照组。通过HE染色观察软腭组织中肌细胞、血管等的变化, 再通过免疫组化检测软腭组织中 HIF-1α 和 VEGF 的表达。 结果 (1)HE染色结果显示:实验组软腭组织中上皮细胞增生,固有层增厚,肌纤维均存在不同程度的变性,肌纤维排列紊乱,新生毛细血管生成等现象;(2)免疫组化染色结果显示: HIF-1α于OSAHS 中、重度组在黏膜基底层及棘层细胞胞浆高表达,阳性表达主要定位于细胞核和细胞浆呈棕褐色。VEGF在OSAHS 中、 重度组软腭组织黏膜鳞状上皮基底层及棘层细胞胞浆强阳性表达,腺管上皮细胞强阳性表达或局灶强阳性表达,血管内皮细胞部分阳性表达;(3)HIF-1α、VEGF均与 睡眠呼吸暂停低通气指数(AHI)呈正相关(P<0. 05), 与夜间最低氧饱和度呈负相关(P<0. 01)。 结论 OSAHS 患者软腭组织中HIF-1α、 VEGF的表达,与OSAHS严重程度有关, 可能在其发生发展过程中起到重要作用。     

Expression of vascular endothelial growth factor and nm23 as prognostic factors in nasopharyngeal carcinoma]

OBJECTIVE: To evaluate the prognostic significance of vascular endothelial growth factor (VEGF) and nm23 expression for patients with nasopharyngeal carcinoma(NPC). METHODS: From Jan. 1991 to Dec. 1995, 75 NPC patients treated in Department of Radiotherapy & Chemotherapy, Zhongnan Hospital of Wuhan University, with follow-up of more than 4 yrs, were selected to enter this study. Specimens of 75 NPC were studied by immunohistochemical staining for VEGF and nm23. The immunohistochemical data were correlated with development of locoregional failure, distant metastasis and survival. RESULTS: Positive expression of VEGF and negative expression of nm23 were found in 54.7% and 56% of NPC specimens. Positive expression of VEGF and negative expression of nm23 correlated with the development of distant metastasis (P = 0.0181; P = 0.0091) and shorter survival (P = 0.0136; P = 0.0207). No association was observed with locoregional failure. No association was observed between the expression of VEGF and the expression of nm23.Cox proportional hazard model was used to compare the various factors and found the VEGF and nm23 expression were significant prognostic factors for patients with NPC. CONCLUSION: The results of the study show that the VEGF and nm23 expression are significant prognostic factors for patients with NPC and indicate that expression of VEGF and nm23 may be useful in identifying patients who may benefit from systemic chemotherapy or other novel treatment strategies.     

Altered expression of inducible nitric oxide synthase (iNOS) in the cochlea

Using immunohistochemistry and Western blot, the expression of inducible nitric oxide synthase (iNOS) in the lateral wall and organ of Corti was examined in normal (unstimulated) and stimulated mice and guinea pigs. The stimuli were: (1). injection of bacterial lipopolysaccharide (LPS, 5 mg/ml) into the middle ear through the tympanic membrane and (2). exposure to a 110 dB SPL (A-weighted) broadband noise, 3 h/day, for three consecutive days. For the unstimulated condition, weak iNOS expression was found in the vascular endothelium, marginal cells, nerve fibers, stereocilia of hair cells and Hensen's cells of the organ of Corti. More intense iNOS fluorescence signals were observed in cochlear tissues (particularly in hair cells and stria vascularis marginal cells) in animals exposed to loud sound or treated with LPS. Although the precise roles of iNOS expression in normal cochlear function have yet to be determined, enhanced iNOS expression following noise exposure and LPS suggests its participation in cochlear pathophysiology, including noise- and inflammatory factor-induced hearing loss.