宫颈癌中MMP-2及TIMP-2的表达及其临床意义

目的 :通过检测基质金属蛋白酶 - 2 (MMP - 2 )及基质金属蛋白酶组织抑制物 - 2 (TIMP - 2 )在宫颈癌中的表达 ,探讨其与宫颈癌侵袭转移的关系。方法 :采用免疫组化S -P法检测 5 1例宫颈癌和 16例正常宫颈组织中MMP - 2和TIMP - 2的表达情况。结果 :MMP - 2、TIMP - 2在正常宫颈上皮组织中均无表达 ,在宫颈癌组织中的阳性表达率分别为 74 .5 % (38/ 5 1)、4 7.1% (2 4 / 5 1) ,有显著性差异 (P <0 .0 1)。MMP - 2、TIMP - 2的表达与组织学类型无关 ,但与临床分期、细胞分化程度、淋巴结转移有关。结论 :MMP - 2、TIMP - 2的表达与宫颈癌的侵袭转移有关 ,MMP - 2、TIMP - 2可作为预测宫颈癌侵袭转移潜能和临床预后的指标。     

MMP9、TIMP1及VEGF在子宫内膜异位症患者在位内膜的表达

研究基质金属蛋白酶 9(matrixmetalloproteinase 9,MMP9)及其抑制剂 (tissueinhibitorofmetalloproteinase 1,TIMP1)和血管内皮生长因子 (vascularendothelialgrowthfactor,VEGF)在子宫内膜异位症 (endometriosis,EM)患者的在位内膜中的表达。采用免疫组化SP法分别检测 4 5例EM(研究组 )在位内膜及 32例子宫肌瘤 (对照组 )的在位内膜中MMP9、TIMP1及VEGF的表达。MMP9、TIMP1、MMP9 TIMP1和VEGF在研究组和对照组的表达分别为 0 336 ,0 2 76 ;0 2 5 3,0 2 6 7和 1 2 93,1 0 34;0 372 ,0 2 88。其中MMP9、MMP9 TIMP1和VEGF在研究组中的表达显著高于对照组内膜 (P <0 0 1)。EM患者的在位内膜中MMP9、VEGF高表达可能是EM发生发展的重要原因。诊刮筛选出高MMP9、VEGF表达的在位内膜可望成为预测和诊断EM的方法之一。     

哮喘豚鼠MMP-9、TIMP-1免疫反应的变化及与气道重塑的关系

目的　观察实验性哮喘豚鼠气道平滑肌基质金属蛋白酶 9(MMP 9)及其抑制剂 (TIMP 1)的表达以及气道平滑肌厚度的变化。方法　用免疫组织化学结合显微图像分析测定气道平滑肌MMP 9、TIMP 1免疫反应的变化及气道平滑肌厚度。结果　MMP 9、TIMP 1广泛分布于气道平滑肌。哮喘组MMP 9平均灰度值(10 5 94± 6 0 9)明显低于对照组 (12 0 2 5± 3 6 6 ) (P <0 0 1) ;哮喘组TIMP 1平均灰度值 (99 36± 5 71)明显低于对照组 (12 3 4 5 7) (P <0 0 1) ;哮喘组气道平滑肌厚度 (6 45± 1 83μm2 / μm)明显高于对照组 (3 17± 0 85 )(P <0 0 5 )。结论　MMP 9、TIMP 1可能参与哮喘时气道重塑。     

垂体腺瘤中E-cadherin和nm23蛋白表达与肿瘤生物学行为的关系

目的探讨垂体腺瘤中Ecadherin(Ecad)和nm23蛋白表达与肿瘤体积、激素分泌和侵袭性的关系。方法应用免疫组化SP法检测Ecad和nm23基因蛋白在23例侵袭性垂体腺瘤和24例非侵袭性垂体腺瘤组织中的表达。结果Ecad和nm23的表达在侵袭性垂体腺瘤组低于非侵袭性组(P<0.05),Ecad表达在非分泌型垂体腺瘤组低于分泌型组且与肿瘤体积呈负相关(P<0.05),Ecad与nm23表达间呈正相关(P<0.05)。结论Ecad与nm23表达降低可能与垂体腺瘤的侵袭性有关,Ecad表达降低可能影响垂体腺瘤细胞分化和促进细胞增殖,Ecad和nm23可作为评估垂体腺瘤侵袭能力的生物学指标之一。     

PTTG基因蛋白的表达与垂体腺瘤侵袭性和增殖能力的关系

目的探讨人垂体腺瘤中垂体肿瘤转化基因(PTTG)蛋白的表达与肿瘤侵袭性和增殖程度的关系。方法采用免疫组织化学染色方法检测手术切除石蜡包埋的63例垂体腺瘤(侵袭组45例,非侵袭组18例)组织中PTTG蛋白的表达,染色增殖细胞核抗原(PCNA),同时计数组织内微血管数量(MVD)。结果 PTTG在侵袭性垂体腺瘤中的表达水平显著高于非侵袭性垂体腺瘤。侵袭性垂体腺瘤中PCNA标记指数和微血管密度也显著高于非侵袭性垂体腺瘤。相关分析显示PTTG表达与垂体腺瘤内PCNA标记指数和微血管密度呈正相关(P<0.05)。结论 PTTG在垂体腺瘤形成过程中起重要作用,并与垂体腺瘤的侵袭性和增殖程度密切相关。     

糖尿病大鼠肾组织中NF-κB和MMP-9及TIMP-1表达关系的研究

目的 :研究糖尿病大鼠肾组织中核因子 κB(NF κB)和基质金属蛋白酶 9(MMP 9)及组织基质蛋白酶抑制因子 1(TIMP 1)表达之间的关系方法 :将 3 0只雄性Wistar大鼠随机分为正常对照组 (C组 )、糖尿病未治疗组 (D组 )和糖尿病苯那普利治疗组 (B组 ) ,每组10只 ,利用链脲佐菌素 (STZ)诱导糖尿病模型 ,应用免疫组化方法研究大鼠肾组织中NF κB、MMP 9和TIMP 1的表达 ,并利用HPIAS 10 0 0型医学彩色图像分析系统进行图像分析。结果 :各组间的差异均有显著性意义 (P <0 .0 1)。NF κB与MMP 9成明显负相关 (r =-0 .882 67,P <0 .0 1) ,NF κB与TIMP 1无明显相关 (r =0 .5 2 981,P >0 .0 5 ) ,NF κB与MMP 9/TIMP 1的比值亦成明显负相关 (r =-0 .8685 0 ,P <0 .0 1)。结论 :NF κB对糖尿病大鼠肾组织中MMP 9的表达起重要作用 ,可能参与了糖尿病肾病的发生和发展     

基质金属蛋白酶及其抑制剂在乳腺癌转移、浸润中的作用

目的 :探讨基质金属蛋白酶 (MMP 2、MMP 9)及其抑制剂 (TIMP 1、TIMP 2 )与乳腺癌的关系。方法 :应用免疫组化S P法检测乳腺癌中MMP 2、MMP 9、TIMP 1、TIMP 2的表达。结果　 30例乳腺癌患者中MMP 2阳性表达 14例 ,占 46 7% ,MMP 9阳性表达 16例占 5 3 3% ,TIMP 1阳性表达 4例 ,占 13 3% ,TIMP 2阳性表达 3例 ,占 10 0 %。MMP 2、MMP 9的表达与乳腺癌的淋巴结状况、肿瘤大小相关 ,与年龄、月经状况无关。结论 :MMP 2、MMP 9的表达可以作为估计乳腺癌预后及术后综合治疗的生物学指标     

缺氧、高氧对肝星形细胞基质金属蛋白酶Ⅱ表达及活性的影响

目的　研究缺氧、高氧对肝星形细胞基质金属蛋白酶Ⅱ (MMP 2 )表达及其活性影响。方法　分离大鼠肝星形细胞 ,在缺氧或高氧条件下培养 ,以免疫细胞化学标记链霉素卵白素生物素(LSAB)法及酶联免疫吸附 (ELISA)法分别检测细胞内MMP 2、基质金属蛋白酶组织抑制因子Ⅱ(TIMP 2 )、膜型基质金属蛋白酶Ⅰ (MT1 MMP)的表达 ,和培养上清中MMP 2、TIMP 2的相对含量 ,并以酶谱法测培养上清MMP 2活力。结果　 (1)缺氧培养 12h ,MMP 2表达增高 (缺氧组阳性指数 :5 7±2 0 ;对照组 :3 2± 1 0 ,;P <0 0 1) ,TIMP 2表达则降低 (缺氧组阳性指数 :2 5± 0 7;对照组 :3 6±1 0 ;P <0 0 5 ) ;培养上清中MMP 2酶活性明显降低 (缺氧组总吸光度值 :7 334± 1 92 2 ;对照组 :17 2 77± 7 4 2 4 ;P <0 0 1)。缺氧不同时段 (6、12、2 4h)比较 ,变化以 6h段最明显。 (2 )高氧培养 12h ,上清中MMP 2、TIMP 2蛋白相对含量均高于对照组 ,TIMP 2更明显 (高氧组A450 :0 0 5 0± 0 0 14 ;对照组 :0 0 2 2± 0 0 10 ;P <0 0 1) ,酶活性亦高于对照组 (高氧组总吸光度值 :5 2 5 2± 0 771;对照组 :4 30 4± 1 0 83;P <0 0 5 ) ,高氧组MT1 MMP表达增强。结论　肝星形细胞对氧敏感。缺氧使肝星形细胞MMP 2表达增加 ,该影响在     

乳腺癌细胞基质MMP-9和TIMP-2的表达及其临床意义

目的 :研究乳腺癌组织中细胞外基质金属蛋白酶 9(MMP 9)和组织基质金属蛋白酶抑制剂 2 (TIMP 2 )的表达及其与淋巴转移和预后的相关性。方法 :通过免疫组织化学的方法检测 10 9例乳腺癌组织中MMP 9和TIMP 2表达的情况。结果 :MMP 9和TIMP 2在人乳腺癌中表达的阳性率分别为 6 6 97%和 30 2 7% ;MMP 9的表达与组织学分级、淋巴结转移呈正相关 (P <0 0 1) ,与术后生存率呈负相关 ,而TIMP 2的表达则相反。结论 :MMP 9和TIMP 2的表达与乳腺癌淋巴结转移与预后密切相关 ,可作为判断乳腺癌转移和预后的指标     

鼻咽癌组织中巨噬细胞移动抑制因子和MMP-2、MMP-9表达的关系

目的　研究巨噬细胞移动抑制因子 (macrophagemigrationinhibitoryfactor,MIF)和MMP 2、MMP 9在鼻咽癌组织中的表达水平及相互关系 ,探讨鼻咽癌细胞早期侵袭转移的机制。方法　收集 4 5例确诊的鼻咽原发癌活检组织标本 ,采用免疫组化LSAB法检测鼻咽癌组织中MIF和MMP 2、MMP 9的表达 ,并分析患者的临床参数的关系。结果　在 4 5例鼻咽原发癌组织中 ,MIF、MMP 2和MMP 9的阳性表达率分别为 77 8% (35 / 4 5 )、6 4 4 % (2 9/ 4 5 )和 71 1% (32 / 4 5 )。其中 ,癌细胞MIF和MMP 9的表达水平均显示与淋巴结转移有关 ,伴有淋巴结转移的癌组织中二者表达水平均高于无淋巴结转移的癌组织 (P值均 <0 0 5 )。MIF阳性组的癌细胞MMP 9的表达 (5 0 2 %± 33 5 % )明显高于MIF阴性病例 (11 7%± 2 2 7% ) ,两者差异有显著性 (P <0 0 1) ,且MIF的表达与MMP 9的表达亦呈正相关 (rs=0 .4 92 ,P <0 0 1) ,但癌细胞MMP 2的表达与MIF、MMP 9的表达以及是否有淋巴结转移则均未显示相关性。以Schmincke型生长方式分布的癌细胞MIF表达水平 (6 7 4 %±35 2 % )也高于以Regaud型方式分布的癌细胞 (32 9%± 2 9 7% ) ,差异有显著性 (P <0 0 1)。结论　鼻咽癌组织中癌细胞的MIF和MMP 9同步过表达 ,可能在鼻咽癌细胞的转移     

Tumor-specific downregulation and methylation of the CDH13 (H-cadherin) and CDH1 (E-cadherin) genes correlate with aggressiveness of human pituitary adenomas.

The gene products of CDH13 and CDH1, H-cadherin and E-cadherin, respectively, play a key role in cell-cell adhesion. Inactivation of the cadherin-mediated cell adhesion system caused by aberrant methylation is a common finding in human cancers, indicating that the CDH13 and CDH1 function as tumor suppressor and invasion suppressor genes. In this study, we analyzed the expression of H-cadherin mRNA and E-cadherin protein in 5 normal pituitary tissues and 69 primary pituitary adenomas including all major types by quantitative real-time RT-PCR (qRT-PCR) and immunohistochemistry, respectively. Reduced expression of H-cadherin was detected in 54% (28/52) of pituitary tumors and was significantly associated with tumor aggressiveness (P<0.05). E-cadherin expression was lost in 30% (21 of 69) and significantly reduced in 32% (22 of 69) of tumors. E-cadherin expression was significantly lower in grade II, III, and IV than in grade I adenomas (P=0.015, P=0.029, and P=0.01, respectively). Using methylation-specific PCR (MSP), promoter hypermethylation of CDH13 and CDH1 was detected in 30 and 36% of 69 adenomas, respectively, but not in 5 normal pituitary tissues. Methylation of CDH13 was observed more frequently in invasive adenomas (42%) than in non-invasive adenomas (19%) (P<0.05) and methylation of CDH1 was more frequent in grade IV adenomas compared with grade I adenomas (P<0.05). Methylation of either CDH13 or CDH1 was identified in 35 cases (51%) and was more frequent in grade IV invasive adenomas than in grade I non-invasive adenomas (P<0.05 and P<0.05, respectively). Downregulation of expression was correlated with promoter hypermethylation in CDH13 and CDH1. In conclusion, the tumor-specific downregulation of expression and methylation of CDH13 and CDH1, alone or in combination, may be involved in the development and invasive growth of pituitary adenomas.     

Analyses of matrix metalloproteinases and their inhibitors in cyst fluid of serous ovarian tumors.

OBJECTIVES: Expression of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) in serous tumors of the ovary was investigated to determine whether and how these proteolytic enzymes are associated with the progression of these tumors. METHODS: Cyst fluid of 24 serous ovarian tumors (8 adenocarcinomas, 2 borderline tumors and 14 adenomas) was analyzed using gelatin/casein zymography and enzyme-linked immunosorbent assay. RESULTS: Concentrations of MMP-9 and MMP-2 were statistically higher in serous adenocarcinomas than in serous adenomas (p < 0.01, p < 0.05, respectively), while the concentrations of TIMP-1 and TIMP-2 showed no significant difference between adenocarcinomas and adenomas. The molar ratio of TIMP-2/MMP-2 was lower in adenocarcinomas than in adenomas (p < 0.05). With gelatin zymography, the MMP-9 band was detected in all serous adenocarcinomas, but only in 8 of 14 serous adenomas (p = 0.05). Using casein zymography, MMP-7 was more frequently detected in serous adenocarcinomas (7/8) than in serous adenomas (4/14; p < 0.05). CONCLUSIONS: These observations indicate that matriolytic enzymes such as MMP-2, MMP-7 and MMP-9 are secreted into cyst fluid from serous adenocarcinoma tissues. In part, the aggressive invasion of serous carcinoma cells may be explained by the expression of matriolytic enzymes.     

Expression of matrix metalloproteinases in benign and malignant follicular thyroid lesions

AIMS: To examine expression of matrix metalloproteinases (MMPs) and related proteins in follicular thyroid lesions (FTLs) and to determine their usefulness for differential diagnosis of FTLs, particularly between minimally invasive carcinoma and adenoma. METHODS AND RESULTS: Six widely invasive follicular carcinomas (WIFCs), 15 minimally invasive follicular carcinomas (MIFCs), 19 follicular adenomas (FAs) and 10 adenomatous goitres (AGs) were analysed immunohistochemically for MMP-1, MMP-2, MMP-7, MMP-9, membrane-type 1-MMP (MT1-MMP) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2). MMP-1 was positive in all FTLs. MMP-2 and MMP-7 were positive in more than 80% of WIFC and MIFC cases, whereas they were negative in all FA and AG cases except one MMP-2+ FA (P < 0.001). MMP-9 stained positive significantly more in MIFC than FA or AG cases (P < 0.05, respectively). The positivity of MT1-MMP and TIMP-2 was different among some of the FTLs, but with no significant difference between MIFC and FA cases. In-situ hybridization of MMP-2 and MMP-7 mRNA in selected cases demonstrated the expression of these enzymes in the tumour cells as well as in some stromal cells. CONCLUSIONS: Our results confirm MMP expression mainly in malignant FTLs and suggest that MMP-2 and MMP-7 may be useful markers to distinguish MIFC from FA.     

IL-19及其受体与慢性鼻-鼻窦炎组织重塑的相关性研究

目的:探讨白细胞介素19(interleukin-19,IL-19)及其受体(IL-20R1/IL-20R2)在不同类型慢性鼻-鼻窦炎[慢性鼻-鼻窦炎伴鼻息肉(chronic rhinosinusitis with nasal polyps,CRSw NP)和慢性鼻-鼻窦炎不伴鼻息肉(chronic rhinosinusitis without nasal polyps,CRSs NP)]患者中表达的差异,并分析其与慢性鼻-鼻窦炎组织重塑的相关性。方法:研究共分3组:CRSw NP组(30例)、CRSs NP组(15例)和对照组(鼻中隔偏曲患者15例)。实时荧光定量PCR检测IL-19及其受体(IL-20R1/IL-20R2)和组织重塑因子基质金属蛋白酶(MMP)-2、MMP-9、金属蛋白酶组织抑制物(TIMP)-1的mRNA相对表达量;免疫组化检测IL-19及其受体的蛋白表达量;ELISA检测MMP-9、TIMP-1的蛋白表达量。结果:CRSw NP组IL-19、IL-20R1、IL-20R2和MMP-9的mRNA和蛋白表达量高于CRSs NP组和对照组(P0.05),CRSs NP组TIMP-1的mRNA和蛋白的表达量均高于CRSw NP组和对照组(P0.05);MMP-2的mRNA和蛋白的表达量在各组无显著差异。CRSw NP组IL-19、IL-20R1和IL-20R2均与MMP-9的mRNA相对表达量呈正相关(P0.05);IL-19及其受体与TIMP-1的mRNA相对表达量无明显相关。结论:IL-19及其受体的mRNA在CRSw NP中高表达,且与MMP-9 mRNA表达呈正相关,提示二者可能存在交互作用,可能参与慢性鼻-鼻窦炎的组织重塑。     

细胞周期抑制蛋白p16、p27蛋白表达与垂体腺瘤侵袭性的关系

目的探讨p27、p16、PCNA的蛋白表达与垂体腺瘤侵袭性的关系,为根治垂体腺瘤及减少肿瘤术后复发提供实验依据。方法应用免疫组化SP法检测43例侵袭性垂体腺瘤和37例非侵袭性腺瘤的p16、p27和PCNA蛋白表达水平,分析p16、p27蛋白表达水平与PCNA蛋白表达之间的相关性。结果垂体侵袭腺瘤p16、p27蛋白表达较非侵袭组明显减低;复发组p16蛋白表达的阳性率与非复发组之间无统计学意义。p27蛋白表达的阳性率较非复发组减低有显著差异。结论p16、p27蛋白表达异常与垂体腺瘤的发生及侵袭性有关,它们的表达情况能作为垂体腺瘤侵袭性的参考指标。     

Increased expression of HMGA1 correlates with tumour invasiveness and proliferation in human pituitary adenomas

Wang E L, Qian Z R, Rahman M M, Yoshimoto K, Yamada S, Kudo E & Sano T
(2010) Histopathology 56, 501–509 Increased expression of HMGA1 correlates with tumour invasiveness and proliferation in human pituitary adenomas Aims: High‐mobility group A1 (HMGA1) is highly expressed in various benign and malignant tumours. The development of pituitary adenoma in Hmga1 transgenic mice has been reported. However, no studies have investigated HMGA1 expression and its clinical significance in human pituitary adenomas. Methods and results: Immunohistochemical expression of HMGA1 was analysed with respect to various clinicopathological factors in 95 pituitary adenomas. Nuclear expression of HMGA1 was observed in 62% of pituitary adenomas, whereas normal adenohypophysial tissues were negative. Although HMGA1 expression was frequently detected in clinically non‐functioning adenomas – 90% of silent adrenocorticotropic hormone (ACTH), 76.2% of follicle‐stimulating hormone/luteinizing hormone and 100% of null cell adenomas – it was also detected in 48.1% of growth hormone (GH), 60% of mixed GH/prolactin (PRL), 62.5% of PRL, 66.6% of thyroid‐stimulating hormone and 37.5% of ACTH adenomas. HMGA1 expression was significantly higher in invasive adenomas or macroadenomas than in non‐invasive adenomas or microadenomas (invasive versus non‐invasive, P < 0.05; macroadenoma versus microadenoma, P < 0.05). In addition, HMGA1 showed the highest level in grade IV, more aggressive pituitary adenomas, than in grades I, II and III (IV versus I, P = 0.01; IV versus II, P = 0.01; IV versus III, P = 0.07). Furthermore, a significant correlation between HMGA1 expression and MIB‐1 labelling index was observed (R = 0.368, P < 0.0002). Conclusions: These findings suggest that HMGA1 up‐regulation has an important oncogenic role in pituitary tumorigenesis, as well as being a novel molecular marker of tumour proliferation and invasiveness.     

PRL-2基因转染对肝细胞基质金属蛋白酶及其抑制剂表达的影响

目的:研究PRL-2基因增强肿瘤细胞侵袭及转移能力的机制。 方法:采用脂质体转染的方法将PRL-2基因表达质粒转染至正常永生化肝细胞系CL1中，G418筛选阳性克隆。应用明胶酶谱法检测转染肝细胞分泌MMPs 酶谱变化，Western blotting 及RT-PCR检测转染肝细胞MMP-2、MMP-9、TIMP-1和TIMP-2的变化。以PRL-2磷酸酯酶特异性抑制剂处理转染细胞，观察抑制PRL-2活性对上述指标的影响。 结果:经过 8周G418筛选及RT-PCR和Western blotting鉴定,获得稳定表达PRL-2的细胞亚系PRL-2-CL1。转染后的CL1细胞分泌MMP－9 、活性型MMP－9 和MMP－2 ,均显著高于转染前CL1细胞的MMPs 分泌(P<0.01);使用特异性抑制剂后， MMP－9 、活性型MMP－9 和MMP－2活性显著降低(P<0.01)。Western blotting及RT-PCR检测显示PRL-2-CL1细胞MMP2、MMP9蛋白及mRNA含量均较转染前显著升高(P<0.05)，TIMP-2则显著降低(P<0.05)；使用抑制剂后，可以逆转上述变化。 结论:PRL-2在永生化肝细胞中获得稳定、高效表达，PRL-2基因增强肝细胞侵袭及转移能力与其提高细胞MMP2、MMP9表达，降低TIMP-2有关。