Immunohistochemical Expression of Matrix Metalloproteinase-7 in Human Colorectal Adenomas Using Specified Automated Cellular Image Analysis System: A Clinicopathological Study

Background/Aim:

To evaluate the immunohistochemical expression of matrix metalloproteinase-7 (MMP-7) in colorectal adenomas, and to correlate this expression with different clinicopathological parameters.

Patients and Methods:

The study was retrospectively designed. Thirty three paraffin blocks from patients with colorectal adenoma and 20 samples of non-tumerous colonic tissue taken as control group were included in the study. MMP-7 expression was assessed by immunohistochemistry method. The scoring of immunohistochemical staining was conducted utilizing a specified automated cellular image analysis system (Digimizer).

Results:

The frequency of positive immunohistochemical expression of MMP-7 was significantly higher in adenoma than control group (45.45% versus 10%) (P value < 0.001). Strong MMP-7 staining was mainly seen in adenoma cases (30.30%) in comparison with control (0%) the difference is significant (P < 0.001). The three digital parameters of MMP-7 immunohistochemical expression (Area (A), Number of objects (N), and intensity (I)) were significantly higher in adenoma than control. Mean (A and I) of MMP-7 showed a significant correlation with large sized adenoma (≥ 1cm) (P < 0.05), also a significant positive correlation of the three digital parameters (A, N, and I) of MMP-7 expression with villous configuration and severe dysplasia in colorectal adenoma had been identified (P < 0.05).

Conclusion:

MMP-7 plays an important role in the growth and malignant conversion of colorectal adenomas as it is more likely to be expressed in advanced colorectal adenomatous polyps with large size, severe dysplasia and villous histology. The use of automated cellular image analysis system (Digmizer) to quantify immunohistochemical staining yields more consistent assay results, converts semi-quantitative assay to a truly quantitative assay, and improves assay objectivity and reproducibility.     

食管鳞癌中基质金属蛋白酶-7的表达及意义

目的:研究基质金属蛋白酶-7(MMP-7)在食管鳞癌中的表达情况及其与肿瘤浸润和淋巴结转移的关系。方法:用免疫组织化学法检测75例食管鳞癌组织及其相应正常粘膜中MMP-7的表达。结果:食管鳞癌组织MMP-7基因表达(59/75,78.7％)高于正常组织(P<0.01)。浸润到外膜层食管鳞癌(37/45,82.2％)与浸润至粘膜下层鳞癌(5/10,50.0％)之间MMP-7的高表达有显著性差异(P<0.05),淋巴结转移组MMP-7高表达(27/29,93.1％)高于淋巴结未转移组(P<0.05)。结论:MMP-7的高表达与食管鳞癌浸润深度和淋巴结转移有关,可能成为食管鳞癌恶性生物学行为的指标。     

Cyclooxygenase-2 Inhibition Prevents Migration of Colorectal Cancer Cells to Extracellular Matrix by Down-Regulation of Matrix Metalloproteinase-2 Expression

Purpose

Matrix metalloproteinases-2 hydrolyses gelatins and collagens. It has many biologic functions, including cancer cells invasion. Overexpression of cyclooxygenase-2 is known to be involved in colorectal carcinogenesis, although the mechanism is unclear. Up-regulation of matrix metalloproteinases-2 expression may be one of the mechanisms, which explains how cyclooxygenase-2 expression promotes migration of colorectal cancer cells to extracellular matrix.

Methods

Colorectal cancer cell lines HT29, CaCO2, and Colo205 were used. By using flow cytometry, their cyclooxygenase-2 expression was determined. These cell lines were modulated with NS398, a selective cyclooxygenase-2-inhibitor, and prostaglandin-E2. Western blot and enzyme-linked inmmunosorbent assay were used to determine these cells’ matrix metalloproteinases-2 expression. These cell lines’ ability to migrate into extracellular matrix was determined by Matrigel® (Millipore, Watford, UK) Invasion Chamber.

Results

HT29 expressed more cyclooxygenase-2 than CaCO2. Cyclooxygenase-2 was not detected in Colo205. Matrix metalloproteinases-2 expression is highest in HT29 and least in Colo205. Cyclooxygenase-2 inhibition by NS398 showed decreased matrix metalloproteinases-2 expression in HT29 and CaCO2, but not Colo205, reversible with prostaglandin-E2. Prostaglandin-E2 was shown to up-regulate matrix metalloproteinases-2 expression in all cell lines. Matrigel® Invasion Chamber demonstrated that many more HT29 cells migrate across the membrane than CaCO2 and Colo205, and cyclooxygenase-2 inhibition reduced cellular migration in the cyclooxygenase-2–positive cell lines. Prostaglandin-E2 promoted migration in all cell lines.

Conclusions

There is a positive relationship between cyclooxygenase-2 and matrix metalloproteinases-2 expression. The latter is modulated by prostaglandin-E2 in all cell lines and NS398 in cyclooxygenase-2–positive cells. Such modulation has a knock-on effect to the cells’ ability to invade into extracellular matrix. Cyclooxygenase-2 and matrix metalloproteinases-2 expression are potential therapeutic targets into prevention of colorectal cancer metastasis.

     

MMP-7、PTEN在子宫内膜增生症及内膜样腺癌中的表达

目的研究基质金属蛋白酶-7(MMP-7)和抑癌基因(PTEN)在子宫内膜增生症至子宫内膜样腺癌组织中的异常表达及变化规律,探讨其作为子宫内膜早期癌变及浸润、转移的生物学标志的可能性。方法应用免疫组化S-P法检测15例正常增生期子宫内膜组织、40例子宫内膜增生症组织、38例子宫内膜样腺癌组织中MMP-7、PTEN的表达。结果在正常增生期子宫内膜,子宫内膜简单型、复杂型、不典型增生及子宫内膜样腺癌,MMP-7的阳性表达率呈递增趋势,PTEN呈递减趋势。等级相关分析结果显示:MMP-7、PTEN表达异常与癌组织学分级均显著相关(r分别为0.410和-0.569,P均<0.01)。除不典型增生外,正常增生期子宫内膜、子宫内膜简单型、复杂型增生与子宫内膜样腺癌组织中的MMP-7、PTEN表达差异有显著性。正常增生期子宫内膜、子宫内膜简单型增生与不典型增生比较,MMP-7、PTEN表达差异显著;在内膜样腺癌中,MMP-7与PTEN表达呈负相关(r=-0.392,P<0.01);MMP-7与肿瘤浸润深度呈正相关(r=0.389,P<0.05);PTEN蛋白表达与子宫内膜样腺癌的临床分期、组织学分级无关(P>0.05),与肌层浸润呈负相关(P<0.05)。结论MMP-7、PTEN蛋白在子宫内膜增生症至子宫内膜样腺癌组织中表达异常,两者可能成为子宫内膜组织早期癌变和预测生物学行为的重要分子标记物。     

基质金属蛋白酶-9在脑缺血中的作用

基质金属蛋白酶-9（MMP-9）是参与细胞外基质降解的重要酶类。研究证实，脑缺血时MMP-9能分解细胞外基质和基底膜，破坏血脑屏障，最终导致神经元凋亡。其表达与脑缺血后血管源性脑水肿的发生、溶栓治疗后的出血性转化、脑白质降解等关系密切，在脑缺血后继发性损伤方面具有重要作用。     

Prospective Observation of Small Adenomas in Patients After Colorectal Cancer Surgery Through Magnification Chromocolonoscopy

Purpose This study was designed to confirm the safety of not removing small adenoma in patients who undergo colorectal cancer surgery. Methods Patients who underwent surveillance colonoscopy after surgery were enrolled. The study was approved by our institutional review board. Colonoscopy was performed with magnification chromocolonoscopy. Benign adenomas of 6 mm or less in size, diagnosed based on both nonmagnified and magnified observation, were left unresected with a maximum of three polyps per patient. The sites of the polyps were marked by tattooing. Interval colonoscopy was performed predominantly yearly or biennially. Increase in size by 2 mm or larger was defined as significant. In follow-up, polyps were removed if they grew larger than 6 mm, were suspicious for high-grade dysplasia, or the patients requested to have polyps removal. Results Five hundred polyps in 284 patients met the above criteria and were not resected, and 412 polyps were followed by repeat colonoscopy. The mean observation period was 3.6 ± 2.2 years and the mean number of repeat colonoscopy was 3.6 ± 1.6. At the final colonoscopy, 71 percent of 412 polyps showed no change in size, 15 percent increased, 3 percent decreased, and 11 percent could not be identified. Eighty-eight polyps were resected endoscopically, and histology showed neither cancer nor adenomas with high-grade dysplasia. Two hundred fifty-five polyps detected in the same patient cohort during index/repeat colonoscopy were removed, including four adenomas with high-grade dysplasia and two T1 cancers. Conclusions Leaving small polyps is safe even in patients who have undergone colorectal cancer surgery, provided that careful observation is guaranteed. Supported in part by Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan. Read at the meeting of The American Society of Colon and Rectal Surgeons, St. Louis, Missouri, June 2 to 6, 2007.     

曲匹地尔对兔主动脉球囊损伤后血管壁基质金属蛋白酶-2表达的影响

目的观察曲匹地尔对球囊损伤后血管内膜增生和基质金属蛋白酶-2(MMP-2)mRNA表达的影响,探讨其抑制内膜增生的可能机制。方法在兔腹主动脉球囊损伤的模型上,光镜下观察曲匹地尔对血管内膜增生的影响及应用逆转录聚合酶链反应的方法观察球囊损伤后MMP-2mRNA的动态表达及曲匹地尔对其表达的影响。结果球囊损伤后1,4,7,14,28d,MMP-2mRNA表达均显著高于对照组,且在损伤后第7天达高峰(1·66±0·14vs对照组0·41±0·14,P<0·01)。曲匹地尔可以抑制血管内膜的增生,并能抑制MMP-2mRNA的表达,损伤后7天抑制作用最为显著(1·15±0·18vs对照组1·66±0·14,P<0·01)。结论球囊损伤后MMP-2的表达可能与血管内膜增生有关。曲匹地尔抑制血管内膜增生的作用可能与其抑制MMP-2的表达有关。     

基质金属蛋白酶-7在哈萨克族食管癌中的表达及其意义

目的 观察哈萨克族食管癌及癌旁组织中基质金属蛋白酶-7的表达,探讨其在哈萨克族食管癌发生、发展,浸润转移中的作用.方法 采用RT-PCR方法检测MMP-7 基因mRNA在20例哈萨克族食管癌组织及其癌旁组织标本中的表达,选取mRNA水平差异组织标本采用Western-bolt方法检测其在蛋白水平的表达情况.结果 20例癌组织中有11例表达阳性,占55%,20例癌旁组织中有8例表达,占40%,其中癌组织表达量明显高于癌旁组织的有8例,占40%;MMP-7蛋白在癌组织中表达高于其相应癌旁组织.结论 MMP-7在哈萨克族食管癌的发生发展中起到一定的作用.     

基质金属蛋白酶-3、C反应蛋白与颈动脉硬化的相关性研究

目的研究颈动脉粥样硬化患者血清基质金属蛋白酶-3(MMP-3)、高敏C反应蛋白(hsCRP)的变化,探讨两者之间的关系,阿托伐他丁的干预作用。方法对77例来院就诊病人行颈动脉超声检查,根据颈动脉内中膜厚度(IMT)、有无动脉斑块及斑块的性质,分为三组:颈动脉软斑组(N=36),颈动脉硬斑组(N=18),对照组(N=23);检测血清MMP-3及hsCRP水平,分析MMP-3及hsCRP的水平与颈动脉硬化、斑块性质的关系,并对其中18例颈动脉软斑患者用阿托伐他丁治疗12周,观察斑块及MMP-3及hsCRP的水平的变化。结果(1)MMP-3水平:颈动脉软斑组、颈动脉硬斑组、对照组分别为8.27±5.12ng/ml、6.15±2.20ng/ml、3.87±2.31ng/ml,三组间差异有统计学意义;CRP水平则分别为4.82±2.02mg/L、1.54±1.01mg/L、1.45±1.01mg/L,颈动脉软斑组高于颈动脉硬斑组及对照组(P<0.001),颈动脉硬斑组与对照组之间差异无统计学意义(P>0.05);(2)颈动脉软斑病人阿托伐丁治疗12周后血MMP-3的水平明显下降,复查颈动脉彩超,斑块明显好转,hsCRP的水平变化不明显(P>0.05)。结论血清MMP-3水平与颈动脉硬化及斑块性质有密切关系,阿托伐他丁有改善颈动脉硬化,降低血清MMP-3水平作用。     

基质金属蛋白酶-9 C-1562T基因多态性与冠心病相关关系的初步研究

目的：了解基质金属蛋白酶-9（matrix metaloproteinase-9，MMP-9）G1562T多态性在山西省人群中的分布。初步探讨其与冠心病（conoary heart disease，CHD）的相关性。方法：采用聚合酶链反应-限制性酶切片段长度多态性（polymerase chain reaction-restriction fragment length polymorphism，PCR-RFLP）方法，判定各研究对象的基因型，并统计各基因型及等位基因的频率。结果：在CHD组（84例）和正常对照组（64例）C／T基因型的频率分别是26．6％、21．4％，T等位基因频率为14．8％、10．7％，两组比较无显著性差异（P值均〉0．05）。将CHD分组后，分别将不稳定心绞痛（unstable angina，UA）（37例）、心肌梗死（myocardial isechemia，MI）（27例）组C／T基因型和T等位基因频率与正常对照组比较，结果表明，UA组（C／T为44．4％，T为25．9％）与正常对照组（C／T为21．4％，T为10．7％）比较差异有统计学意义（均P〈0．05）。结论：CHD组中UA患者MMP-9C1562TC／T基因型及T等位基因频率与正常对照组比较差异有统计学意义（P〈O．05）。结合临床资料分析，CHD组C／T和T／T基因型即T等位基因携带者65岁以下患者占61％，且多为不稳定心绞痛患者（72．7％）．提示了MMP-9C-1562T多态性可能是65岁以下UA患者的一个危险因素。     

大鼠实验性脑出血后基质金属蛋白酶2和9的表达

目的探讨大鼠脑出血后基质金属蛋白酶2和基质金属蛋白酶9的表达及其意义。方法采用未抗凝新鲜自体股动脉血注入大鼠尾状核建立脑出血动物模型。用酶谱分析法测定脑出血后6h、12h、24h、3天、6天、10天和15天时基质金属蛋白酶2和基质金属蛋白酶9的表达。结果基质金属蛋白酶2在脑出血后24h开始表达(P<0.05),第6天时达高峰(P<0.01),到第10天仍维持高水平表达(P<0.05);而基质金属蛋白酶9在脑出血后6h开始表达(P<0.05),24h~3天最明显(P<0.01),此后活性逐渐降低,第10天时的活性仍高于对照组(P>0.05),15天时不再表达。结论脑出血后6h~3天出血灶周围脑组织中基质金属蛋白酶9的活性增高,可能与急性期脑水肿的形成有关,而基质金属蛋白酶2的迟发表达可能参与了脑组织的修复过程。     

Cyclooxygenase-2 Expression in Colorectal Adenomas

PURPOSE: Cyclooxygenase-2 is an important target for nonsteroidal anti-inflammatory drugs in suppressing colorectal tumorigenesis. To evaluate the role of cyclooxygenase-2 in sporadic colorectal adenoma, we correlated cyclooxygenase-2 expression in adenomas with other adenoma characteristics. METHODS: Cyclooxygenase-2 expression was evaluated immunohistochemically in 95 endoscopically resected colorectal adenomas. RESULTS: Cyclooxygenase-2 was expressed mainly in the cytoplasm of adenoma cells, where it was seen in 74 percent (70/95) of adenomas. Expression was related significantly to grade of dysplasia (P < 0.001) and tumor size (P = 0.028). Multivariate logistic regression analysis showed cyclooxygenase-2 expression in adenoma cells to be independently associated with grade of dysplasia (P = 0.001). CONCLUSION: Observed associations suggest that cyclooxygenase-2 plays an important role in progression of the adenoma-to-carcinoma sequence.     

基质金属蛋白酶-9与缺血性脑血管病

基质金属蛋白酶-9与缺血性脑血管病的发生发展、脑缺血后损伤和再灌注、继发性出血等关系密切，控制基质金属蛋白酶-9水平有助于改善缺血性脑血管病的防治。     

基质金属蛋白酶9在动脉粥样硬化中的研究进展

基质金属蛋白酶9(MMP-9)是一种主要表达于巨噬细胞的含锌离子的蛋白酶,参与了细胞外基质的合成与降解、炎症介质的调控,可促进动脉粥样硬化的发生及进展、血管壁的重构,导致严重的心血管事件发生。本文将重点讨论MMP-9在动脉粥样硬化中的研究进展,并就其在急性心肌梗死的预测价值和心肌梗死后心脏重塑方面加以探讨。     

血管生成素-2与基质金属蛋白酶-7在大肠癌中的表达及意义

目的:研究血管生成素-2(Ang-2)及基质金属蛋白酶-7(MMP-7)蛋白在人大肠癌组织中的表达及其与大肠癌临床病理特征的关系．方法:应用免疫组化法检测40例大肠癌及其癌旁正常组织中Ang-2及MMP-7蛋白表达水平．结果:大肠癌组织中Ang-2蛋白表达阳性率为77．5％(31/40),明显高于癌旁正常组织 40％(16/40)(P=0．001);Ang-2表达水平与患者性别、年龄及癌组织的部位、大小、分化程度、淋巴结转移无关(P>0．05);与浸润深度, 远处转移及Dukes’分期相关(分别为P=0．007, P=0．023,P=0．008);大肠癌组织RMMP-7蛋白表达阳性率为85％(34/40),明显高于癌旁正常组织35％(14/40)(P=0．000)．MMP-7蛋白表达阳性率与Ang-2的表达相关(P=0．016)．结论:Ang-2蛋白可能通过促进MMP-7的表达从而促进了大肠癌的生长及转移．     

基质金属蛋白酶-9、C反应蛋白与2型糖尿病患者颈动脉粥样硬化的相关性研究

目的:探讨基质金属蛋白酶-9(MMP-9)、C反应蛋白(CRP)与2型糖尿病颈动脉粥样硬化的关系, 方法:用酶联免疫法测定50例单纯2型糖尿病患者(A组)及50例2型糖尿病合并颈动脉粥样硬化患者(B 组)的血清MMP-9、CRP水平,通过彩色多普勒超声测量颈动脉内中膜厚度、斑块及血管狭窄情况判断颈动脉粥样硬化。结果:B组血清MMP-9、CRP水平分别为600.30 ng/ml、5.98 mg/L,较A组血清MMP-9、CRP水平284.05 ng/ml、3.46 mg/L明显增高,两组相比均有极显著性差异(P<0.01)。调整混杂因素后B组血清MMP一9、CRP水平仍较A组高(369.52 ng/ml比153.16 n/ml及4.03 mg/ml比2.19 mg/L),且两组相比有极显著性差异(P<0.01)。结论:MMP-9、CRP在2型糖尿病大血管病变的发生发展中起着重要作用。