壶腹癌MMP-9、TIMP-1的表达及其与淋巴结转移的相关性研究

目的：探讨基质金属蛋白酶9（MMP－9）、组织金属蛋白酶抑制因子－1在壶腹癌中表达的意义。方法：采用免疫组化S－P法对32例壶腹癌组织进行MMP－9、TIMP－1的检测。结果：MMP－9、TIMP－1在壶腹癌组织中阳性表达率分别为65．6％（21／32），46．9％（15／32），MMP－9的表达与壶腹癌患者淋巴结转移呈正相关（P＜0．05），而TIMP－1的表达则相反，MMP－9和TIMP－1的表达呈负相关（P＜0．01）。结论：MMP－9可作为判断壶腹癌恶性生物学行为的标记物。人工诱导TIMP－1的表达可能抑制肿瘤的浸润和转移，为肿瘤治疗提供了新的方向。     

基质金属蛋白酶9及组织型基质金属蛋白酶抑制因子1在大鼠外伤性脑水肿中的表达及意义

目的 探讨大鼠脑损伤中基质金属蛋白酶9(MMP9)及组织型基质金属蛋白酶抑制因子1(TIMP1)的表达变化规律及意义.方法 采用Feeney自由落体法制作大鼠轻度脑损伤模型,采用干湿重法测定脑组织含水量;采用逆转录-聚合酶链反应(RT-PCR)法对大鼠脑损伤模型中MMP9及TIMP1 mRNA进行检测并分析两者比值.结果 实验组脑组织含水量(BWC),MMP9mRNA及TIMP1 mRNA表达量及两者比值均明显高于对照组,P值分别为0.013、0.000、0.000及0.011,差异有统计学意义(P<0.05),MMP9 mRNA与TIMP1mRNA表达量及两者比值与BWC均呈正相关,r值分别为0.654、0.355(P<0.05).结论 MMP9/TIMP1的比例失衡可能在外伤性脑水肿的发生发展过程中起重要作用.     

肝门部胆管癌中MMP-2/TIMP-2比值半定量研究的临床意义

目的 探讨MMP-2和TIMP-2在肝门部胆管癌浸润、转移过程中的作用及其与预后的关系,为判断肿瘤恶性程度、浸润转移、预后风险及提高临床治疗效果提供实验依据。方法 采用免疫组化法结合计算机图象分析软件技术检测肝门部胆管癌中MMP-2和TIMP-2的表达情况,将临床结合相关病理参数进行分析及统计学处理。结果 MMP-2的表达肝内有浸润组高于肝内无浸润组,TIMP-2的表达相反。图象定量分析结果显示:二者阳性染色强度指数比值与肿瘤临床分型、局部淋巴结有无转移、有无肝内浸润转移、肿瘤组织分化程度和生存期有关(P<0.05);与肿瘤组织学类型无关(P>0.05)。结论 MMP-2有促进肿瘤浸润转移功能,TIMP-2具有抗肿瘤浸润转移功能,临床上检测MMP-2/TIMP-2比值有助于肝门部胆管癌生物学行为的判断和预后风险的估计。     

基质金属蛋白酶(MMP)9、MMP2 mRNA及基质金属蛋白酶抑制剂2 mRNA在人膀胱移行细胞癌中的表达及意义

目的探讨膀胱移行细胞癌中基质金属蛋白酶（MMP）9、MMP2及基质金属蛋白酶抑制剂2（TIMP2）mRNA表达与膀胱肿瘤恶性程度和侵袭性之间的关系及其临床意义。方法采用逆转录-聚合酶链反应（RT-PCR）方法检测36例膀胱移行细胞癌及5例正常膀胱组织MMP9、MMP2、TIMP2mRNA的表达。结果MMP9、MMP2mRNA随着肿瘤分级分期的增高，表达明显增强[x^-值（MMP9）G1：1．218，G2：1．681，G3：1．811；T2～T4：1．840，Tis～T1：1．399；P〈0．01；（MMP2）G1：1．323，G2：1．694，G3：2．060；T2～T4：1．950，Tis-T1：1．505；P〈0．01]，且明显高于正常膀胱组织中的表达[互值（MMP9）0．455，（MMP2）0．461；P〈0．01]。TIMP2mRNA随肿瘤分级的增高而明显减少（x^-值：G1：1．489，G2：1．391，G3；0．580；P〈0．01）。结论（1）MMP9、MMP2mRNA的高表达与BTCC的恶性程度呈正相关；（2）TIMP2mRNA的表达与BTCC的恶性程度呈负相关。     

细菌性颅内动脉瘤72-和92-KDa IV型胶原酶及其组织抑制因子mRNA的原位杂交

目的　探讨细菌性颅内动脉瘤 (BIAs)的发病机理。方法　用地高辛标记的 72 和92 KDaIV型胶原酶 (MMP 2和MMP 9)及其组织抑制因子 (TIMP) 1、TIMP 2反义RNA探针与BIAs(n =2 )和正常脑动脉组织 (n =6 )进行原位杂交 ,定位产生它们的细胞。并用正义RNA探针进行阴性对照。结果　 2份感染性颅内动脉瘤标本中均见MMP 9mRNA阳性杂交信号。且在内膜 ,尤其是相当于内弹力层部位 ,阳性杂交信号密集存在。动脉瘤壁内弹力层消失。阳性杂交信号定位于单核细胞、成纤维细胞和平滑肌细胞。正常脑动脉标本中未发现MMP 9mRNA阳性杂交信号。在BIAs和正常脑动脉组织中未发现MMP 2、TIMP 1和TIMP 2的mRNA阳性杂交信号。结论　感染因素引起炎性细胞浸润 ,表达MMP 9mRNA增多 ,导致内弹力层断裂、消失 ,甚至动脉壁全层结构的破坏 ,从而引发BIAs形成。     

肝癌组织中β-葡萄糖醛酸酶mRNA与临床病理因素的关系

目的 探讨肝癌组织中β-葡萄糖醛酸酶(β-glucuronidase,β-G)mRNA的表达与临床病理因素的关系.方法 采用RT-PCR法检测25例肝癌组织及10例正常肝组织中β-G mRNA的表达,并分析肝癌组织中β-G mRNA与临床病理因素的关系.结果 患者的年龄、肿瘤大小、AFP水平不影响肝癌组织中β-G mRNA的表达.β-G mRNA的表达在有门静脉癌栓以及淋巴结转移者中明显高于无门静脉癌栓以及淋巴结转移者(t=7.857,9.341,P＜0.05).结论 β-G mRNA的表达与肝癌患者有无门静脉癌栓及淋巴结转移等临床病理因素密切相关,β-G可能在肝癌细胞的侵袭和转移过程中起一定的作用.     

Metalloproteinases and their inhibitors: influence on tumor invasiveness and metastasis formation in head and neck squamous cell carcinomas

BACKGROUND: Matrix metalloproteinases (MMPs) play an important role in tumor invasiveness. This study investigates the expression status of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in head and neck squamous cell carcinomas (HNSCC). METHODS: Of 48 laryngeal squamous cell carcinoma (LSCC) biopsies and 10 HNSCC cell lines, mRNA was isolated, reversely transcribed, and subjected to polymerase chain reaction (PCR) amplifying MMP-1, MMP-2, MMP-9, MMP-10, TIMP-1, and TIMP-2. Silver nitrate-stained gel electrophoresis demonstrated MMP and TIMP expression status. Exemplary immunohistochemistry and zymography confirmed translation and enzyme activity. RESULTS: Densitometric analysis revealed MMP-2 expression and lymph node metastases to be positively and TIMP-1 and TIMP-2 to be negatively correlated with lymph node metastases. TIMP-2 expression and tumor size were negatively correlated. MMP-1, MMP-9, and MMP-10 expression were not correlated to metastasis formation or tumor size. CONCLUSIONS: Our results suggest that MMP-2 expression enhances, whereas TIMP-1 and TIMP-2 both suppress, cancer spread in LSCC.     

基质金属蛋白酶-9和血管内皮生长因子与原发性肝癌侵袭转移关系研究

目的　探讨原发性肝癌中MMP 9和VEGF基因的表达 ,探讨其与原发性肝癌侵袭转移的关系。方法　采用逆转录 聚合酶链式反应 (RT PCR)技术 ,检测 41例原发性肝癌手术切除肿瘤组织、癌旁组织标本的MMP 9和VEGF基因表达。结果　肝癌组织和癌旁 1cm、5cm组织中分别有 70 7%、48 9%、41 5 %MMP 9表达阳性和 75 6%、5 3 7%、46 3 %VEGF表达阳性 ,癌组织的MMP 9和VEGF表达水平显著高于癌旁 5cm组织 (P <0 0 1) ,癌组织与癌旁 1cm组织MMP 9和VEGF表达水平无显著性差异。癌组织中MMP 9和VEGF的表达水平与有无包膜、门静脉癌栓有显著性差异 (P <0 0 5 )。MMP 9与复发、AFP阳性有显著性差异 (P <0 0 5 ) ,而VEGF与之无差异。结论　MMP 9和VEGF基因的过度表达与肝癌侵袭转移密切相关 ,MMP 9可作为判断预后的指标     

转移相关蛋白与乳腺癌转移潜能的相关性分析

目的:探讨与肿瘤转移相关的蛋白在乳腺浸润性导管癌中的表达及其意义。方法:应用组织芯片技术及SP免疫组化方法检测247例乳腺浸润性导管癌中α B晶体蛋白，CD44v6， MMP 2,TIMP 2的表达，并分析它们与乳腺癌临床病理特征的关系。结果:(1) 在浸润性导管癌中, α B晶体蛋白，CD44v6，MMP 2,TIMP 2的阳性表达率分别为70.0%，61.5%， 57.5%,57.1%；与正常组织比较差异有统计学意义（P＜0.001）；（2）单因素分析显示α B晶体蛋白，CD44v6,MMP 2的表达与淋巴结转移呈正相关（r=0.317，0.623,0.705），而TIMP 2的表达与之呈负相关(r=-0.532)；(3）多因素分析显示淋巴结转移与α B晶体蛋白，CD44v6，MMP 2,TIMP 2的表达相关，该4项指标联合对淋巴结转移的预测率达89.5%。结论:α B晶体蛋白，CD44v6， MMP 2,TIMP 2的表达情况与乳腺浸润性导管癌的转移密切相关，通过对原发瘤多种转移相关蛋白的检测可以预测乳腺癌的转移能力。     

Plasma concentrations of metalloproteinases (MMP-1, MMP-3) and their inhibitors in patients with prostate cancer

Summary Matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) are involved in important processes of tumor invasion and metastasis. In the study presented, matrix metalloproteinase 1 (MMP1) and 3 (MMP3), the tissue inhibitor of metalloproteinase 1 (TIMP1) and the complex MMP1/TIMP1 were measured by ELISA tests specific for these proteins in blood plasma. These components have been investigated in prostate cancer patients (PCa) with metastases (n = 18; T2, 3, 4 pN1, 2M1), prostate cancer patients without metastases (n = 29; T2, 3 pN0M0), patients with benign prostate hyperplasia (BPH; n = 29) and in healthy men (n = 35). Mean values of MMP1 and of the complex MMP1/TIMP1 were not different among the four groups. The mean values of MMP3 and especially TIMP1 were significantly higher in prostate cancer patients with metastases compared with controls, BPH patients and prostate cancer patients without metastases. Ten of these 18 patients had TIMP1 levels higher than the upper reference limit. TIMP1 concentrations correlate to the tumor stage but not to the tumor grade. These results indicate, that TIMP1 could be an potential marker for metastases in prostate cancer patients.      

次级淋巴组织趋化因子在胃癌淋巴结转移中的作用

目的：研究次级淋巴组织趋化因子（secondary lymphoid tissue chemokine,SLC)在胃癌淋巴结中的表达，以进一步阐明胃癌淋巴转移的机制。方法：用逆转录聚合酶链反应（RT－PCR）测定29例胃癌组织淋巴结中SLC mRNA的表达。结果：胃癌转移淋巴结中SLC mRNA的表达均显著低于相应的正常胃黏模组织，但其表达与病人的临床病理特征并不相关。结论：在胃癌转移淋巴结中，SLC的表达明显受抑，后者可能在胃癌的淋巴结转移中起了重要作用，提高淋巴结中SLC的表达可能成为控制胃癌转移和复发的又一关键途径。     

去甲斑蝥素对荷瘤裸鼠胆囊癌移植瘤的抗癌作用机制

目的 探讨去甲斑蝥素（NCTD）对荷瘤裸鼠胆囊癌移植瘤的抗癌作用机制。方法 在建立裸鼠胆囊癌移植瘤动物模型的基础上进行肿瘤增殖、侵袭、转移体内干预实验,实验分空白对照、氟尿嘧啶、NCTD、NCTD＋氟尿嘧啶4组。6周末应用链霉素亲和生物素复合物（SABC）法检测移植瘤增殖细胞核抗原（PCNA）、Ki-67、细胞周期素D1（cyclin D1）、p27、Bcl-2蛋白,逆转录PCR（RT-PCR）检测PCNA、cyclin D1、p27、Bcl-2、Bax、存活素（Survivin）基因mRNA。HE染色观察瘤周癌细胞浸润、侵袭,解剖显微镜下观察肺组织转移瘤结节,并采用SABC和RT-PCR检测nm23、金属蛋白酶2（MMP2）及其组织抑制剂（TIMP2）蛋白和mRNA。结果 NCTD组增殖相关PCNA、Ki-67、cyclin D1蛋白表达下降,p27蛋白表达上升;PCNA mRNA、cyclin D1 mRNA表达下降,p27 mRNA表达增高。NCTD组凋亡相关Bcl-2蛋白表达下降;Bcl-2 mRNA、Survivin mRNA表达下降,Bax mRNA表达增高。NCTD显著减少荷瘤鼠移植瘤的瘤周癌细胞浸润和肺转移结节（P〈0．01）;NCTD组转移相关MMP2蛋白表达下降,nm23、TIMP2蛋白表达上升;nm23-H,mRNA、TIMP2 mRNA表达增高。结论 NCTD抑制荷瘤裸鼠胆囊癌移植瘤增殖、侵袭和转移的机制可能与NCTD干扰胆囊癌移植瘤细胞周期,抑制细胞增殖,诱导细胞凋亡,阻止细胞迁移运动,以及影响细胞增殖、细胞周期调控、细胞凋亡、细胞基质溶解和转移相关基因蛋白表达有关。     

纤溶酶原激活物抑制物1与肝细胞癌

目的研究纤溶酶原激活物抑制物１（ＰＡＩ１）在肝细胞癌（ＨＣＣ）蛋白和ｍＲＮＡ水平的表达及其与ＨＣＣ生物学特性的关系。方法取ＨＣＣ石蜡标本４８例,肝良性肿瘤石蜡标本１２例（对照组）做免疫组化染色;液氮冻存ＨＣＣ标本２０例,肝血管瘤５例（对照组）做免疫印迹杂交。结果肝癌细胞与癌周细胞及对照组肝细胞相比,ＰＡＩ１抗原蛋白和ｍＲＮＡ表达显著升高,差异有显著意义,Ｐ值分别＜００１和＜０．０５。术后２年内死亡病例与生存病例相比,ＰＡＩ１阳性率有显著意义的升高,Ｐ＜００５。ＰＡＩ１和纤溶酶原激活物（ｕＰＡ）及其受体（ｕＰＡＲ）同时阳性患者与同时阴性患者相比,前者侵袭性病例较后者升高有显著性意义（Ｐ＜００５）。结论ＨＣＣ中ＰＡＩ１蛋白和ｍＲＮＡ表达明显增高。ＰＡＩ１与ＨＣＣ浸润转移和预后密切相关。     

促血管生成素在肝癌组织中的表达与肝癌新生血管形成及转移的关系

目的　观察促血管生成素 (angiopoietin)在肝细胞癌组织中的表达 ,探讨其临床意义。方法　采用原位分子杂交和免疫组织化学染色方法检测两种促血管生成素Angiopoietin 1(Ang 1)、Angiopoietin 2 (Ang 2 )以及血管平滑肌激动蛋白 (α smoothmuscleactin ,α SMA)在正常肝脏组织和肝细胞癌组织中的表达。结果　Ang 1mRNA在正常肝脏和肝癌组织中都有表达 ,肝癌组织和癌旁组织 (5 6± 6 /HP)以及正常肝脏 (6 7± 11/HP)之间差异没有显著意义 (t=3 2 14 ,P =0 12 6 )。Ang 2mRNA在正常肝脏不表达 ,肝癌组织中阳性表达 (阳性细胞数为 32± 8/HP)。Ang 2集中于癌灶边缘、血管周围。肝癌组织中微血管密度 (15± 2 /HP)和新生血管的数量 (9± 3/HP)明显高于正常肝脏(分别为 5± 1/HP和阴性 ) ,Ang 2的表达随肝癌组织的病理学分级、有无转移以及新生血管数和微血管密度而增加 (t=2 714 ,P =0 0 31)。结论　促血管生成素在肝癌的新生血管形成和转移过程中可能起到一定作用。Ang 2可能参与了肝癌的新生血管形成。     

淋巴结微转移及相关蛋白检测在大肠癌患者Dukes分期、治疗和预后中的意义

目的 探讨淋巴结微转移（LNMM）和nm23-H1、基质金属蛋白酶9（MMP9）、金属蛋白酶2组织抑制因子（TIMP2）蛋白检测及其相关性在大肠癌患者Dukes分期、治疗和预后中的意义。方法 应用免疫组化SABC法检测30例DukesB期大肠癌淋巴结细胞角蛋白20（CK20）和癌组织nm23-H1、MMP9、TIMP2蛋白表达,另对同期30例DukesC和D期大肠癌患者检测nm23-H1、MMP9和TIMP2;随访、记录患者的临床病理参数和生存资料,分析其相关性。结果 （1）26．7％DukesB期大肠癌患者、7．8％DukesB期大肠癌淋巴结存在CK20阳性。（2）DukesB期大肠癌nm23-H1、MMP9表达与DukesC和D期差异显著（P〈0．05）;nm23-H,表达下降和（或）MMP9表达增强与LNMM相关（P〈0．05）,两者预测大肠癌LNMM敏感性和特异性分别为62．5％和81．8％、75．0％和69．8％,联合检测特异性则达90．9％;而TIMP2与Dukes分期、LNMM无关。（3）DukesB期LNMM（＋）患者癌复发转移率明显高于同期LNMM（-）组（P〈0．05）,而生存率则降低（P〈0．05）;nm23-H1（-）LNMM（＋）、MMP9（＋）LNMM（＋）患者生存期明显短于nm23-H1（＋）LNMM（-）、MMPq（＋）LNMM（-）组（P〈0．05）。结论 CK20免疫组化可检出大肠癌LNMM;DukesB期大肠癌nm23-H1、MMP9表达与LNMM相关,且表达异常LNMM患者预后差;联合检测淋巴结CK20和癌组织rim23-H1、MMP9表达,对大肠癌Dukes分期、术后辅助化疗和预后判断有重要意义。     

Diabetes-related intestinal region-specific thickening of ganglionic basement membrane and regionally decreased matrix metalloproteinase 9 expression in myenteric ganglia

BACKGROUNDThe importance of the neuronal microenvironment has been recently highlighted in gut region-specific diabetic enteric neuropathy. Regionally distinct thickening of endothelial basement membrane (BM) of intestinal capillaries supplying the myenteric ganglia coincide with neuronal damage in different intestinal segments. Accelerated synthesis of matrix molecules and reduced degradation of matrix components may also contribute to the imbalance of extracellular matrix dynamics resulting in BM thickening. Among the matrix degrading proteinases, matrix metalloproteinase 9 (MMP9) and its tissue inhibitor (TIMP1) are essential in regulating extracellular matrix remodelling.AIMTo evaluate the intestinal segment-specific effects of diabetes and insulin replacement on ganglionic BM thickness, MMP9 and TIMP1 expression.METHODSTen weeks after the onset of hyperglycaemia gut segments were taken from the duodenum and ileum of streptozotocin-induced diabetic, insulin-treated diabetic and sex- and age-matched control rats. The thickness of BM surrounding myenteric ganglia was measured by electron microscopic morphometry. Whole-mount preparations of myenteric plexus were prepared from the different gut regions for MMP9/TIMP1 double-labelling fluorescent immunohistochemistry. Post-embedding immunogold electron microscopy was applied on ultrathin sections to evaluate the MMP9 and TIMP1 expression in myenteric ganglia and their microenvironment from different gut segments and conditions. The MMP9 and TIMP1 messenger ribonucleic acid (mRNA) level was measured by quantitative polymerase chain reaction.RESULTSTen weeks after the onset of hyperglycaemia, the ganglionic BM was significantly thickened in the diabetic ileum, while it remained intact in the duodenum. The immediate insulin treatment prevented the diabetes-related thickening of the BM surrounding the ileal myenteric ganglia. Quantification of particle density showed an increasing tendency for MMP9 and a decreasing tendency for TIMP1 from the proximal to the distal small intestine under control conditions. In the diabetic ileum, the number of MMP9-indicating gold particles decreased in myenteric ganglia, endothelial cells of capillaries and intestinal smooth muscle cells, however, it remained unchanged in all duodenal compartments. The MMP9/TIMP1 ratio was also decreased in ileal ganglia only. However, a marked segment-specific induction was revealed in MMP9 and TIMP1 at the mRNA levels.CONCLUSIONThese findings support that the regional decrease in MMP9 expression in myenteric ganglia and their microenvironment may contribute to extracellular matrix accumulation, resulting in a region-specific thickening of ganglionic BM.     

Role of gelatinases (matrix metalloproteinases 2 and 9), vascular endothelial growth factor and endostatin on clinicopathological behaviour of rectal cancer

Aim The aim of this study was to evaluate the role of matrix metalloproteinases (MMPs), their tissue inhibitors [tissue inhibitors of metalloproteinases (TIMPs)] and activators [membrane‐type MMPs (MT1‐MMPs)], vascular endothelial growth factor (VEGF) and endostatin on clinicopathological variables and prognosis in patients with rectal cancer. Method Paired samples of tumour tissue and normal tissue were obtained from patients with rectal cancer who underwent curative surgery (n = 34). Gelatin zymography for MMP‐2 and MMP‐9, an activity assay for MT1‐MMP and enzyme‐linked immunoassays for TIMP‐2, VEGF and endostatin were performed using extracts from the paired tissue samples. Results Active MMP‐9 showed statistically significant relationships with metastatic disease and perineural invasion (P = 0.002 and P = 0.042). A significant relationship was observed between the levels of tumoral pro‐MMP‐2 and pro‐MMP‐9 and the presence of lymph node metastasis (P = 0.012 and P = 0.021, respectively). Tumoral TIMP‐2 levels showed a significant relationship with tumour recurrence (P = 0.011). A significant relationship was also observed between tumour VEGF levels and the presence of perineural invasion (P = 0.044), and VEGF levels were correlated with the size of the tumour (P = 0.009, r = 0.454). Conclusion These results might contribute to further investigation of a possible prognostic significance in rectal cancer.