MMP—9和TIMP—2在喉癌中的表达及其临床意义

目的：探讨基质金属蛋白酶（matrix metalloproteinase,MMP）中的MMP－9及其组织抑制因子－2（tissue inhibitor of metallorpoteinase,TIMP）在喉癌中的表达及其临床意义。方法：采用免疫组织化学法检测29例喉癌石蜡包埋组织中的MMP－9和TIMP－2的表达。结果：MMP－9在喉癌组织中阳性表达率为79．3％，TIMP－2阳性表达率为89．6％，MMP－9与TIMP－2强阳性表达率差异明显（P＜0．05），MMP－9在晚期喉癌中表达增高，T1，T2与T3，T4期喉癌强阳性表达率差异显著（P＜0．05），但弱阳性表达率与临床分期无关，而TIMP－2在喉癌组织中的强阳性表达率与弱阳性表达率均与临床分期有关。MMP－9在淋巴结转移组的强阳性表达率明显高于非淋巴结转移组（P＜0．05），而TIMP－2淋巴结转移强阳性表达率则低于非淋巴结转移组。结论：MMP－9与TIMP－2与喉癌的浸润和淋巴结转移有关，MMP－9与TIMP－2两者关系密切，二者平衡的破坏是喉癌侵袭与转移的重要因素。     

基质金属蛋白酶-11和基质金属蛋白酶-7与喉癌浸润转移及预后的相关性

目的　检测基质金属蛋白酶-11（MMP-11）和基质金属蛋白酶-7（MMP-7）在喉癌组织中的表达,探讨其与喉癌浸润和转移及预后的关系。方法　采用免疫组化方法  检测63例喉癌和癌旁组织中MMP-11和MMP-7蛋白水平表达。结果　①癌组织的阳性表达率明显高于癌旁正常黏膜组织;②MMP-11、MMP-7蛋白表达在T3+T4组高于T1+T2组,淋巴结转移组高于非淋巴结转移组;③Kaplan-Meier生存曲线显示：MMP-11、MMP-7蛋白表达强阳性组预后较阴性、弱阳性组差。结论　MMP-11、MMP-7的表达与喉癌浸润、转移及预后密切相关,可作为预测喉癌浸润和转移潜能及评估预后参考指标之一。     

高迁移率族蛋白B1和基质金属蛋白酶-2、9在喉癌组织中的表达及其与预后的关系

目的:探讨喉癌组织中高迁移率族蛋白B1(HMGB1)和基质金属蛋白酶(MMP)-2、MMP-9的表达及其与临床病理特征和预后的关系。方法:应用免疫组织化学EnVision二步法检测61例喉癌组织中HMGB1和MMP-2、MMP-9蛋白表达;实时荧光定量PCR法检测30例喉癌组织中HMGB1和MMP-2、MMP-9mRNA表达。结果:HMGB1和MMP-2、MMP-9蛋白在喉癌组织中的阳性表达率明显高于癌旁正常组织(χ2值分别为44.934和49.923、36.054,均P<0.01);HMGB1和MMP-2、MMP-9mRNA在喉癌组织中的相对表达水平明显高于癌旁正常组织(t值分别为5.940和7.005、7.664,均P<0.01);HMGB1和MMP-2、MMP-9蛋白表达均与喉癌T分期、临床分期和淋巴结转移有关(P<0.05或<0.01);HMGB1和MMP-9蛋白在喉癌组织中的表达呈正相关(r=0.381,P<0.01),HMGB1和MMP-2蛋白在喉癌组织中的表达无相关性(r=0.094,P>0.05);Kap-lan-Meier生存分析结果显示,HMGB1和MMP-9表达阳性患者的生存率低于表达阴性患者(χ2值分别为4.974、6.418,均P<0.05);COX回归分析显示,HMGB1是危险因子,其表达越高,患者预后越差(P<0.05)。结论:HMGB1和MMP-2、MMP-9与喉癌的浸润、转移有关;HMGB1和MMP-9的表达有协同促进作用;HMGB1可能是影响预后的独立危险因素。     

喉鳞癌中NO、iNOS的测定及其临床意义

目的 :探讨一氧化氮 (NO)、诱生型一氧化氮合酶 (i NOS)在喉鳞癌中的表达及其临床意义。方法 :采用硝酸还原酶法测定健康人和喉癌患者血清 NO3和 NO- 2 之和 ;采用逆转录 -聚合酶链反应 (RT- PCR)技术检测 4 0例不同病理分化程度、临床分期的喉鳞癌组织、癌旁组织及 10例喉乳头状瘤、7例正常喉粘膜中诱生型一氧化氮合酶 (i NOS) m RNA的表达。结果 :喉鳞癌患者血清 NO含量显著高于正常对照组 ,喉癌组织 i NOS m RNA阳性表达率为 85 .0 0 % ,较对照组癌旁组织 (2 0 .0 0 % )、良性肿瘤组 (40 .0 0 % )、正常人组 (0 / 7)明显增高 (P <0 .0 0 0 1,P <0 .0 0 5 ,P <0 .0 0 0 1) ,有淋巴结转移组 (19/ 19)显著高于无淋巴结转移组 (71.4 2 % ) (P<0 .0 5 ) ,i NOS m RNA阳性表达与肿瘤原发部位及 T分级无关 ,与细胞分化程度呈负相关。结论 :喉鳞癌组织中存在 i NOS,它可能通过合成一氧化氮 (NO)在分子水平参与了喉癌的发生、发展     

声门上喉癌MMP-2和E-cadherin的表达与颈部淋巴结转移的相关性研究

目的:探讨MMP-2、E-cadherin在喉癌组织中的表达及其与喉癌颈部淋巴结转移之间的关系。方法:应用免疫组织化学SP法检测10例正常喉黏膜和48例原发性声门上喉癌组织中MMP-2和E-cadherin的表达。结果:MMP-2在声门上喉癌组织中的表达明显高于正常喉黏膜组织中的表达;淋巴结转移组明显高于无淋巴结转移组(P<0.05);E-cadherin在正常喉黏膜组织中的表达明显高于在声门上喉癌组织中的表达;在淋巴结转移组明显低于无淋巴结转移组(P<0.05)。MMP-2和E-cadherin在声门上喉癌组织中的表达呈负相关(r=-0.41)。分别用MMP-2( )、E-cadherin(-)及MMP-2( )与E-cadherin(-)联合来预测喉癌颈部淋巴结转移,MMP-2( )的敏感性最高(91.7%),特异性(58.3%)和阳性预测值(68.8%)最低。MMP-2( )与E-cadherin(-)联合的敏感性最低(79.2%),特异性(95.8%)和阳性预测值(95.0%)最高。E-cadherin(-)的各项指标介于两者之间。结论:MMP-2和E-cadherin蛋白的表达可以作为判断声门上喉癌淋巴结转移的有效指标,联合应用MMP-2和E-cadherin蛋白的检测能够提高判断声门上喉癌淋巴结转移的准确率,对于声门上喉癌的手术治疗具有指导意义。     

人喉鳞状细胞癌中PTEN、PCNA的表达及临床意义

目的　研究抑癌基因PTEN(phosphotaseandtensionhomologdeletedfromchromsome 10 )、增殖细胞核抗原PCNA(proliferatingcellnuclearantigen)在人喉鳞状细胞癌中的表达及临床意义。 方法　应用免疫组化S -P法检测 10例正常喉组织 ,2 0例癌旁组织和 6 0例喉鳞状细胞癌中抑癌基因PTEN ,增殖细胞核抗原PCNA的表达。结果　 10例正常喉组织和 2 0例癌旁组织均有较强的PTEN蛋白的表达 ,15 % (9/ 6 0 )的喉癌呈PTEN蛋白阴性 ,31.7% (19/ 6 0 )的喉癌呈PTEN蛋白弱阳性 ,5 3.3% (32 / 6 0 )的喉癌呈PTEN蛋白阳性或强阳性 ;PTEN阴性喉癌颈部淋巴结转移率为 77.8% (7/ 9) ;PTEN阳性喉癌颈部淋巴结转移率为 33.3% (17/ 5 1) ,两者之间差异有统计学意义 (P <0 .0 5 )。 83.3% (5 0 / 6 0 )的喉癌呈PCNA阳性表达 ,显著高于正常及癌旁组织 (P <0 .0 1)。晚期、低分化、伴颈部淋巴结转移的喉癌组织中PCNA阳性表达率显著高于早期、高分化、无颈部淋巴结转移的喉癌组织(P <0 .0 1)。结论　抑癌基因PTEN的表达异常可能与喉鳞状细胞癌的发生、发展有关 ;PCNA在喉癌中的表达与其分级、临床分期、淋巴结转移密切相关 ,PCNA可作为喉癌进展的重要参考指标。     

西安及深圳地区鼻咽癌组织中MMP-9和TIMP-1的表达及意义

目的:探讨鼻咽癌组织中基质金属蛋白酶9(MMP-9)和基质金属蛋白酶组织抑制剂-1(TIMP-1)的表达在陕西西安与深圳地区之间是否存在地域差别及其意义。方法:应用免疫组织化学方法分别检测西安地区21例、深圳地区22例鼻咽癌组织中MMP-9和TIMP-1表达的情况。结果:西安与深圳地区鼻咽癌组织中MMP-9阳性表达率分别为61.90%和68.18%;TIMP-1阳性表达率两地区分别为28.57%和31.82%。两地区鼻咽癌组织中MMP-9与TIMP-1表达均有显著负相关性。西安地区MMP-9表达与淋巴结转移有显著关系,TIMP-1则与淋巴结转移无相关性。两地区之间MMP-9、TIMP-1的表达差异无统计学意义(P>0.05)。结论:鼻咽癌组织中MMP-9、TIMP-1的表达在西安与深圳两地区之间无差别;MMP-9高表达在鼻咽癌浸润转移中有重要作用。     

转移相关基因CD44V6在甲状腺癌中的表达及其意义

目的 :研究转移相关基因 CD44 V6 在甲状腺癌组织中的表达及其临床意义。方法 :应用免疫组化 SP方法测定45例甲状腺癌组织中的 CD44 V6 的表达阳性率 ,分析其阳性率与肿瘤转移的关系。结果 :CD44 V6 表达阳性率在淋巴结转移组 (85 .0 % )明显高于无淋巴结转移组 (48.0 % ) (P<0 .0 5 ) ;在乳头状腺癌中的表达率为 75 .0 % ,滤泡腺癌组为 6 8.8% ,在髓样癌中及未化分癌中未见阳性表达 ;45～ 6 0岁病人组 CD44 V6 表达阳性率高于其它年龄段。分析 CD44 V6 阳性表达与临床分期的关系 ,发现随分期增加而增加 ,CD44 V6 的表达阳性率在 、 、 期分别为 6 4.3% ,77.8% ,10 0 .0 % ;死亡的 8例中有 6例 CD44 V6 表达阳性。结论 :CD44 V6 表达与甲状腺癌的颈淋巴结转移密切相关 ;与临床分期、年龄、预后及病理类型有一定关系。     

CD44v6在甲状腺乳头状腺癌中的表达及意义

目的 :探讨肿瘤转移相关基因 CD44v6与甲状腺乳头状腺癌颈淋巴结转移及其生物学行为的关系。方法 :采用免疫组化 L SAB方法对 5 0例甲状腺乳头状腺癌存档标本进行 CD44v6表达测定。结果 :甲状腺乳头状腺癌颈淋巴结转移组 2 6例 CD44v6阳性表达率为 76 .9% (2 0 /2 6 ) ,颈淋巴结转移阴性组 2 4例中阳性率为 5 0 .0 % (1 2 /2 4) ,二者比较 P <0 .0 5 ;CD44v6表达阳性率与病人临床 T分期无明显相关性 (P >0 .0 5 )。结论 :CD44v6表达与甲状腺乳头状腺癌的颈淋巴结转移密切相关 ,对预测颈淋巴结转移的危险性有一定参考价值。     

MMP-7和MMP-14在喉癌中的表达及临床意义

目的 研究基质金属蛋白酶7(matrix metallo-proteinase-7,MMP-7)及MMP-14在喉癌中的表达及临床意义.方法 采用免疫组化方法检测60例喉癌组织、34例癌旁组织、22例正常喉黏膜组织中MMP-7和MMP-14的表达,统计分析其在标本中的表达及相关性.结果 ①MMP-7和MMP-14在喉癌组织、癌旁组织及正常喉黏膜组织中阳性和强阳性表达率3组间两两比较,喉癌组织与癌旁组织、正常喉黏膜组织差异均具有统计学意义(P均<0.01);癌旁组织与正常喉黏膜组织差异无统计学意义(P>0.05);②MMP-7和MMP-14强阳性表达与颈淋巴结转移、临床分期及组织病理学分级具有显著相关性(P均<0.05);⑨MMP-7和MMP-14在喉癌组织中的阳性表达率呈正相关(r=0.969,P<0.05).结论 ①MMP-7和MMP-14的高表达与喉癌的发病、浸润、转移有关;②MMP-7和MMP-14两者在喉癌组织中的阳性率呈显著正相关,联合检测对喉癌的治疗和预后判断具有临床意义.     

Matrix metalloproteinase-2 and Matrix metalloproteinase-9 in cholesteatoma and deep meatal skin

A qualitative and quantitative study of the presence of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9) in cholesteatoma was performed. Ten cholesteatoma and four deep meatal skin specimens were analysed for gelatinase activity at molecular weights corresponding to MMP-2 and MMP-9 using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE) Zymography. Gelatinase activity at 72 kDa and 92 kDa was investigated. Western blotting was employed using primary monoclonal antibodies to provide a qualitative assessment of MMP-2 and MMP-9. Non-parametric data analysis using the Mann–Whitney U test did not show a significant difference in expression of MMP-2 (P = 0.51) or MMP-9 (P = 0.14) between the two tissue types. Western blotting showed the presence of both MMP-2 and MMP-9 in the majority of specimens, both cholesteatoma and deep meatal skin.     

Matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 gene expressions and their differential regulation by proinflammatory cytokines and prostaglandin in nasal polyp fibroblasts.

Chronic inflammation of the paranasal sinus leads to nasal polyp (NP) formation. In this study, we investigated the effect of stimulation of the proinflammatory cytokines interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha) and prostaglandin (PG) E2 on the production of messenger RNA (mRNA) of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-I (TIMP-1) in nasal polyp fibroblasts (NPFs) and nasal mucosa fibroblasts (NFs). The mRNAs of IL-1alpha, TNF-alpha, MMP-1, and TIMP-1 in 40 surgical specimens of NPs were studied by in situ hybridization to corroborate the in vitro findings. The results indicated a significant amount of constitutive MMP-1 mRNA in NPFs and cytokine-induced MMP-1 steady-state mRNAs in NFs. The effect of stimulation of cytokines on TIMP-1 mRNA synthesis was unremarkable in NPFs and NFs. Exogenous PGE2 enhanced cytokine-stimulated MMP-1 mRNA synthesis in NPFs. In situ hybridization revealed that cells expressing MMP-1 and TIMP-1 mRNAs (primarily plasma cells, fibroblasts, and endothelial cells) gathered around areas with loose stroma, suggestive of rapid extracellular matrix degradation. These data suggest that the pathogenesis of nasal polyposis could be related to production of MMP-1 and consequent promotion of matrix collagenolysis.     

Distribution and relative activity of matrix metalloproteinase-2 in human coronal dentin

The presence of matrix metalloproteinase-2 (MMP-2) in dentin has been reported, but its distribution and activity level in mature human coronal dentin are not well understood. The purpose of this study was to determine the MMP-2 distribution and relative activity in demineralized dentin. Crowns of twenty eight human molars were sectioned into inner (ID), middle (MD), and outer dentin (OD) regions and demineralized. MMP-2 was extracted with 0.33 mol·L -1 EDTA/2 mol·L -1 guanidine-HCl, pH 7.4, and MMP-2 concentration was estimated with enzyme-linked immunoabsorbant assay (ELISA). Further characterization was accomplished by Western blotting analysis and gelatin zymography. The mean concentrations of MMP-2 per mg dentin protein in the dentin regions were significantly different (P=0.043): 0.9 ng (ID), 0.4 ng (MD), and 2.2 ng (OD), respectively. The pattern of MMP-2 concentration was OD>ID>MD. Western blotting analysis detected ～66 and ～72 kDa immunopositive proteins corresponding to pro-and mature MMP-2, respectively, in the ID and MD, and a ～66 kDa protein in the OD. Gelatinolytic activity consistent with MMP-2 was detected in all regions. Interestingly, the pattern of levels of Western blot immunodetection and gelatinolytic activity was MD>ID>OD. The concentration of MMP-2 in human coronal dentin was highest in the region of dentin that contains the dentinoenamel junction and least in the middle region of dentin. However, levels of Western blot immunodetection and gelatinolytic activity did not correlate with the estimated regional concentrations of MMP-2, potentially indicating region specific protein interactions.     

黏着斑激酶与基质金属蛋白酶9在垂体腺瘤中的表达

目的研究黏着斑激酶（focal adhesion kinase,FAK）和基质金属蛋白酶9（matrix metalloproteinase 9,MMP-9）在垂体腺瘤中的表达及意义。方法垂体腺瘤组织标本50例,其中侵袭性27例,非侵袭性23例,利用免疫组化法检测并分析比较FAK及MMP-9在侵袭性垂体腺瘤和非侵袭性垂体腺瘤中的表达。结果侵袭性垂体腺瘤中FAK和MMP-9的表达均高于非侵袭性垂体腺瘤（P〈0.05）;在侵袭性垂体腺瘤中FAK和MMP-9的表达水平呈正相关（P〈0.05）。结论垂体腺瘤的侵袭性与FAK、MMP-9的过度表达有关,FAK和MMP-9可作为辅助诊断侵袭性垂体腺瘤的一项生物学指标。     

Study of matrix metalloproteinase-2 and -9 in thyroid papillary cancer

Metastasis of cancer starts with the penetration of cancer cells through the membrane surrounding the cancer focus into the stroma (extracellular matrix). The focal membrane consists of mainly type-IV collagen. An immunochemical study of 28 patients with benign thyroid nodular diseases and 27 patients with papillary carcinoma revealed the fragmentation of type-IV collagen in 4 patients with papillary carcinoma. Matrix metalloprotease (MMP)-2 and MMP-9 are the major enzymes which decompose type-IV collagen, and they have been suggested to be related to cancer metastasis. Therefore, we conducted biochemical and immunohistochemical studies to determine the relationship between these MMPs and the degree of malignancy in thyroid diseases. The concentration of MMP-2 in the serum of patients with papillary carcinoma and patients with benign nodules was 526.0 +/- 96.6 and 522.7 +/- 114.6 ng/ml, respectively, and that of MMP-9 was 53.8 +/- 40.3 and 39.9 +/- 36.0 respectively. There was no significant difference between the two groups in the concentration of either enzyme. The concentration of TIMP-2 in the serum was below the detectable level. On the other hand, the concentration of MMP-2 in the tissue of papillary carcinoma, benign nodules and normal tissue was 12.1 +/- 8.1, 5.7 +/- 4.3, and 0.6 +/- 0.5 ng/mg tissue protein, respectively, and that of MMP-9 was 4.2 +/- 4.1, 2.1 +/- 1.7, and 0.4 +/- 0.3 ng/mg tissue protein, respectively. Concentrations of both enzymes were significantly higher in the papillary carcinoma tissue. Immunohistochemical studies revealed a diffuse granular distribution of MMP-2 in the cytoplasm of the tumor cells. These findings imply that MMP-2 and MMP-9 are related to the degree of malignancy of cancer, especially metastasis.     

基质金属蛋白酶2抑制物在鼻咽癌中的表达及与其生物学行为的关系

目的:研究基质金属蛋白酶2抑制物(TIMP2)在鼻咽癌中的表达及其与鼻咽癌生物学行为的关系.方法:采用免疫组织化学 Elivision Plus二步法对50例鼻咽癌组织、15例炎性鼻咽黏膜组织TIMP2进行检测,并进行χ2检验.结果:免疫组织化学结果显示TIMP2在鼻咽癌组织中表达显著高于炎性鼻咽黏膜组织(P<0.05).TIMP2在有淋巴结转移者极显著高于无淋巴结转移者(P<0.01).TIMP2随临床浸润分期升高其表达上调,但差异无统计学意义(P>0.05).结论:表明TIMP2在鼻咽癌细胞突破基底膜向外扩散及转移中起着重要作用,MMP2/TIMP2比率失衡在判断鼻咽癌侵袭性、估计其预后中有一定意义.     

Matrix metalloproteinase-2 and -9 expression in sinonasal inverted papilloma

STATEMENT OF PROBLEM: Inverted papilloma (IP) is a proliferative lesion of the epithelium lining the sinonasal tract, characterized by marked propensity for recurrence and association with carcinoma. To determine a putative role of matrix metalloproteinase-2 (MMP-2) and MMP-9 in the establishment of IP, their expression was studied in IP. METHODS: Archived surgical specimens from 15 IPs were studied using immunohistochemistry and compared to 12 nasal polyps (NP), a model of chronic respiratory mucosal inflammation, and to 6 control nasal mucosa (CM) samples obtained from snorers during turbinectomy. Within IP, MMP-2 and -9 expression was compared between tumoral areas with hyperplastic epithelium and non tumoral areas with nonhyperplastic epithelium. RESULTS: In IP, MMP-2 and MMP-9 epithelial expression was not different compared to CM and NP. MMP-9 expression in submucosal inflammatory cells was not different between IP and CM or NP. However, within IP, a significantly increased number of MMP-9 positive inflammatory cells in the lamina propria adjacent to the hyperplastic epithelium was observed compared to the lamina propria adjacent to nonhyperplastic epithelium. CONCLUSION: Our findings suggest that MMP 9 expressing inflammatory cells may be involved in the pathophysiology of IP.