骨巨细胞瘤基质金属蛋白酶系统的表达及其与复发的关系

目的：研究骨巨细胞瘤组织中基质金属蛋白酶-2和基质金属蛋白酶-9（matrix metalloproteinase,MMP-2,MMP-9)及其特异性抑制剂金属蛋白酶组织抑制物-1和金属蛋白酶组织抑制物-2（tissue inhibitors of metalloproteinase,TIMP-1和TIMP-2）和尿激酶型纤溶蛋白酶原激活物（urokinase type plasminogen activator,uPA)的表达与肿瘤复发的关系。方法：用免疫组化法和原位杂交法检测45例骨巨细胞瘤组织中MMP2、MMP-9、TIMP-1、TIMP-2和uPA蛋白和RNA的表达,计算MMP-2/TIMP-2和MMP-9/TIMP-1的比值,对比肿瘤复发组和非复发组的差异。结果：骨巨细胞瘤组织有高水平的MMP-2、MMP-9、TIMP-1、TIMP-2和uPA表达,但与肿瘤的形态分级无关。复发组MMP-2、MMP-9和MMP-2/TIMP-2、MMP-9/TIMP-1比值以及uPA表达均比非复发组明显增高,高水平MMP-2、MMP-9、MMP2/TIMP-2、MMP-9/TIMP-1表达的病例复发时间较早,复发风险较高。结论：骨巨细胞瘤的复发与基质金属蛋白酶和酶激活物表达增高,酶抑制剂表达减低相关。     

外周血基质金属蛋白酶及其抑制剂与甲状腺癌相关性分析

目的:探讨外周血中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、金属蛋白酶组织抑制剂-2(TIMP-2)和金属蛋白酶组织抑制剂-1(TIMP-1)与甲状腺癌的相关性。方法:采用酶联免疫吸附法(ELISA)和RT-PCR法检测30例甲状腺癌患者、27例良性甲状腺肿瘤患者和25例健康志愿者外周血中MMP-2、MMP-9、TIMP-2和TIMP-1的表达水平,分析两者与甲状腺癌之间的关系。结果:甲状腺癌、良性甲状腺病变和健康对照组血清中MMP-2的表达分别为(118.88±7.59)、(51.13±6.89)和(47.55±8.61)ng/mL,TIMP-2分别为(70.78±8.58)、(36.05±7.34)和(32.56±7.27)ng/mL,MMP-9分别为(110.18±15.49)、(56.39±10.90)和(49.28±11.64)ng/mL,TIMP-1分别为(63.62±9.95)、(38.53±8.31)和(34.01±8.22)ng/mL。甲状腺癌患者外周血中的表达均明显高于良性甲状腺肿瘤患者和健康志愿者P=0.000;良性甲状腺肿瘤患者和健康志愿者之间差异无统计学意义,P>0.05。结论:外周血中MMP-2、MMP-9、TIMP-2和TIMP-1的表达水平有助于甲状腺肿瘤良恶性的鉴别,是重要的术前评估手段。     

MMPs及TIMPs在脑膜瘤脑浸润和瘤周水肿中的作用

目的：研究基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)及其特异性抑制剂金属蛋白酶组织抑制剂-1(TIMP-1)、金属蛋白酶组织抑制剂-2(TIMP-2)在脑膜瘤组织中的表达与肿瘤脑浸润和瘤周水肿及组织学类型的关系。方法：用免疫组化分别检测脑膜瘤组织中MMP-2、MMP-9、TIMP-1和TIMP-2蛋白的表达；并比较各指标在不同组织中表达的差异性，分析其意义。结果：有脑浸润或瘤周水肿的脑膜瘤MMP-2、MMP-9阳性表达率较无脑浸润，无瘤周水肿的脑膜瘤高(P=0.023、0.028，P=0．027、0．031)；非典型性及恶性脑膜瘤较良性脑膜瘤高表达MMP=2、MMP=9，P=0．023、0.025；TIMP-1、TIMP-2的表达阳性率在各肿瘤中差异无统计学意叉，P=0．065、0.075；脑浸润与瘤周水肿关系密切(χ^2=11．23，P=0.0024)。结论：MMP-2、MMP-9、TIMP-1、TIMP-2对脑膜瘤的生物学行为的评估有重要意义。     

基质金属蛋白酶及其组织抑制物在子宫内膜癌中的表达

目的研究基质金属蛋白酶(MMP)-9及其组织抑制物(TIMP)-1在子宫内膜癌组织中的表达及其临床病理学意义.方法应用链霉素抗生物素蛋白-过氧化物酶免疫组织化学方法对40例子宫内膜癌组织及10例正常子宫内膜组织中MMP-9及TIMP-1蛋白及活性进行检测.结果子宫内膜癌组织中MMP-9及TIMP-1的阳性率高于正常子宫内膜,差异有显著性.组织学分级为G3的子宫内膜癌组织中MMP-9和TIMP-1的表达水平高于G2和G1,手术病理分期Ⅱ～Ⅲ期的子宫内膜癌组织中MMP-9和TIMP-1的表达水平高于Ⅰ期.结论子宫内膜癌组织中MMP-9和TIMP-1的表达随着组织学分级和手术病理分期的升高而增加.     

肺癌MMPs和TIMPs的表达及浸润转移的研究

目的 研究肺癌组织中基质金属蛋白晦(MMPs)及其抑制剂(TIMPs)任肺癌组织中的表达情况，以探讨MMPs及TIMPs在肺癌浸润和转移中的作用。方法用SP免疫组化技术检测MMP1、MMP2、MMP-9、MMP-13、TIMP-1、TIMP-2在104例肺癌组织中的表达。结果肺癌组织中MMPs、TIMPs的阳性表达率均显著高于正常肺组织。MMP-2与肺癌分级有密切关系，MMP9与淋巴结转移有密切关系，TIMP与肺癌分期和淋巴结转移有密切关系。肺癌组织中MMP-2与其抑制剂TIMP-2的表达呈正相关，MMP-9与TIMP-2的表达呈正相关。结论MMPs和TIMPs在肺癌发展过程中具有重要作用，尤其是MMP-9和TIMP-1在肺癌的浸润转移过程中具有促进作用。     

白藜芦醇对宫颈癌HeLa细胞基质金属蛋白酶及其组织抑制剂的影响

目的探讨白藜芦醇对宫颈癌侵袭和转移的影响及其初步作用机制。方法选用Transwell侵袭系统,观察白藜芦醇对宫颈癌细胞侵袭转移的影响;用明胶酶谱法、RT-PCR和Western blot法了解白藜芦醇对宫颈癌细胞MMP-2、MMP-9、TIMP-1、TIMP-2的活性、mRNA及蛋白表达的影响。结果白藜芦醇1、0.5mmol/L两组平均穿过的细胞数分别为9.20±1.924,10.20±2.049与空白对照组(39.80±2.387)相比,明显减少,P<0.05。同时,白藜芦醇能够明显降低宫颈癌细胞MMP-2、MMP-9的活性、mRNA及蛋白表达水平(P<0.05),上调TIMP-1、TIMP-2的活性(F=636.354,P<0.05,F=87.516,P<0.05)、mRNA(F=9.000,P<0.05,F=10.288,P<0.05)及蛋白表达水平(F=39.329,P<0.05,F=12.148,P<0.05),并使得MMP-2/TIMP-2、MMP-9/TIMP-1减小,P<0.05。结论白藜芦醇能有效抑制宫颈癌侵袭和转移。其作用机制可能与降低宫颈癌细胞运动能力、抑制MMP-2、MMP-9的活性、mRNA及蛋白表达水平,上调TIMP-1、TIMP-2的活性、mRNA及蛋白表达水平,并使得MMP-2/TIMP-2、MMP-9/TIMP-1减小,MMP-2/TIMP-2、MMP-9/TIMP-1的平衡能力相应提高有关。     

血浆MMPs水平与甲状腺乳头状癌浸润转移的关系

背景与目的:甲状腺乳头状癌是常见的甲状腺恶性肿瘤,目前尚无理想的血液学指标来反映其生物学特性,本研究观察血浆基质金属蛋白酶(MMP)-2、-9活性与甲状腺乳头状癌侵袭和转移的关系。方法:温州医学院附属第二医院1999～2003年共有经病理确诊、行甲状腺癌改良根治术的甲状腺乳头状癌患者74例,用明胶酶谱法测定患者血浆MMP-2和MMP-9活性。结果:淋巴结转移组的血浆活化MMP-2和MMP-9水平(71.9±11.7、15.5±6.1)明显高于无淋巴结转移组(35.2±6.6、7.3±2.3)(P<0.001);包膜及包膜外浸润组的血浆活化MMP-2和MMP-9水平(70.5±13.0、14.7±6.1)明显高于包膜内浸润组(35.4±7.9、8.0±4.2)(P<0.001);而肿瘤直径≥1cm组和<1cm组血浆活化MMP-2和MMP-9水平(55.4±20.4vs.59.3±20.8,10.8±5.7vs.13.7±6.8)无显著性差异(P>0.05),各组之间血浆总MMP-2和MMP-9水平无显著性差异(P>0.05)。结论:明胶酶的活化形式MMP-2和MMP-9的血浆水平与甲状腺乳头状癌的转移、侵袭有关,与肿瘤的大小无关。     

重组人血管内皮抑制素注射液与多西紫杉醇不同顺序用药的抗肿瘤效应观察

目的 观察重组人血管内皮抑制素注射液(商品名:恩度)与多西紫杉醇联合使用时不同给药顺序的抗肿瘤效应.方法 建立肺腺癌A549荷瘤裸鼠模型,随机分为3组,每组8只.(1)同时用药组:每只小鼠恩度400μg/d,第1～35天;多西紫杉醇10 mg/kg,第1～19天,每3 d给药1次.(2)先恩度组:每只小鼠恩度400 μg/d,第1～35天;多西紫杉醇10 mg/kg,第16～34天,每3 d给药1次.(3)模型组:每只小鼠生理盐水100 μl/d,第1～35天;注射用水200 μl/d,第1～35天,每3 d注射1次.同时设立空白对照组(未荷瘤的正常裸鼠,8只),注射方法同模型组.实验过程中测量各组裸鼠体重、移植瘤体积,并计算抑瘤率.实验结束后,采用流式细胞术检测裸鼠外周血循环内皮细胞(CECs)数,采用免疫组化法检测移植瘤组织中基质金属蛋白酶2(MMP-2)、MMP-9、MMP-2的抑制剂(TIMP-2)、MMP-9的抑制剂(TIMP-1)、细胞外MMP诱导因子(EMMPRIN)、α-平滑肌肌动蛋白(SMA)的表达情况,并计数微血管密度(MVD).结果 同时用药组肿瘤体积增长为39.94 mm3,先恩度组肿瘤体积增长为(99.57±74.48)mm3,二者均明显小于模型组[(217.67±95.44)mm3,均P＜0.05].同时用药组、先恩度组、模型组和空白对照组裸小鼠外周血中CECs的数量分别为(25.86±11.77)个/104个细胞、(77.25±24.02)个/104个细胞、(14.71±11.07)个/104个细胞和(12.90±11.20)个/104个细胞,同时用药组、模型组和空白对照组均明显低于先恩度组(均P＜0.01).同时用药组和先恩度组移植瘤组织中MMP-2和MMP-9的表达均较模型组下调(均P＜0.05),先恩度组移植瘤组织中TIMP-1和同时用药组TIMP-2的表达均较模型组上调(均P＜0.05),同时用药组EMMPRIN的表达较模型组下调(P＜0.05).同时用药组和先恩度组移植瘤组织的MVD及α-SMA的水平均低于模型组(均P＜0.05).结论 同时用药组的抑瘤效果及小鼠生存质量更好.同时用药组和先恩度组均可通过下调MMPs的表达以限制肿瘤进展.CECs的变化可能是先上升后下降的动态过程,早期升高可能提示新生血管床面积减小,继而下降则反映联合治疗后的CECs凋亡和肿瘤消退,动态观察CECs的变化可能有助于了解肿瘤负荷及其血管床的变化并预测疗效.     

基质金属蛋白酶MMP-9、2及其抑制物TIMP-1、2在侵袭性垂体腺瘤生物学行为中的意义

背景与目的:通常垂体腺瘤在组织学上属良性肿瘤,但仍有部分肿瘤侵犯周围组织,形成侵袭性肿瘤,垂体腺瘤侵袭的生物学机制尚不清楚。垂体腺瘤血管丰富,而血管形成与肿瘤侵袭都需要细胞外基质成份的降解和细胞移动,这一过程中基质金属蛋白酶(matrixmetalloproteinases,MMP)和它们的天然抑制物-组织金属蛋白酶抑制剂(tissueinhibitorsofmetalloproteinases,TIMP)有可能起着关键作用。本研究拟通过检测MMP-9和MMP-2及其抑制因子TIMP-1和TIMP-2在非侵袭性垂体瘤和侵袭性垂体瘤中的表达,探讨垂体腺瘤的侵袭机制。方法:收集垂体瘤手术切除标本61例,分为侵袭组(49例)和非侵袭组(12例)。采用免疫组织化学SP法检测,并根据阳性细胞占肿瘤细胞总数的比率进行半定量非参数检验分析。结果:MMP-9、TIMP-1、MMP-2和TIMP-2在侵袭性垂体瘤中表达的阳性率分别为95.9%(47/49),57.1%(28/49),75.5%(37/49),和89.8%(44/49);在非侵袭性垂体瘤中阳性率分别为100%(12/12),91.7%(11/12),66.7%(8/12)和91.7%(11/12)。TIMP-1和TIMP-2的表达在侵袭性垂体瘤组明显少于非侵袭性垂体瘤组,(P<0.05);MMP-2在侵袭性垂体瘤组有增加的趋势,但无统计学意义,(P>0.05)。结论:在垂体腺瘤的侵袭性生物学机制中组织金属蛋白酶抑制剂TIMP-1和TIMP-2有可     

血浆MMPs活性检测对甲状腺乳头状癌患者的临床意义

目的本研究观察血浆基质金属蛋白酶MMP-2和MMP-9活性与甲状腺乳头状癌侵袭和转移的关系,以探讨其临床意义.方法本院1999年～2003年共有明确病理结果、无基础疾病史并经检查排除相关疾病的甲状腺乳头状癌患者74例,所有患者行甲状腺癌改良根治术(肿瘤侧腺叶全切加夹部切除加对侧次全切,颈部淋巴结清扫),术后依照病理结果分组,并用明胶酶谱法测定患者血浆MMP-2和MMP-9活性.检测结果采用SPSS10.0 for windows软件包进行t检验.结果淋巴结转移组的血浆MMP-2(62 KD)和MMP-9(82 KD)活性(71.93±11.72、15.50±6.06)明显高于无淋巴结转移组(35.16±6.63、7.27±2.30)(P＜0.001);包膜及包膜外浸润组的血浆MMP-2(62 KD)和MMP-9(82 KD)活性(70.46±13.04、14.67±6.13)明显高于包膜内浸润组(35.43±7.92、8.04±4.23)(P＜0.001);而肿瘤直径≥1cm组和＜1cm组血浆MMP-2(62 KD)和MMP-9(82 KD)活性(55.43±20.38 vs 59.30±20.77,10.85±5.72 vs 13.71±6.77)无显著差别(P＞0.05),各组之间MMP-2(62 KD+72 KD)和MMP-9(82 KD+92 KD)活性无显著差别(P＞0.05).结论明胶酶的活化形式MMP-2(62 KD)和MMP-9(82 KD)和甲状腺乳头状癌转移与侵袭有关,和肿瘤的大小无关,可作为术前评估的重要指标.     

Quercetin inhibits the invasion of murine melanoma B16-BL6 cells by decreasing pro-MMP-9 via the PKC pathway

Purpose: On the basis of the inhibitory effect of quercetin on the invasion of melanoma B16-BL6 cells previously reported by us, the mechanisms of quercetin-mediated inhibition of invasion were further investigated in the present study.Methods: The ability of B16-BL6 cells to invade and migrate was evaluated in terms of the numbers of cells penetrating a reconstituted basement membrane in the Transwell coculture system. The relative levels and activities of matrix metalloproteinase-9 (MMP-9) and MMP-2 were determined by gelatin zymography and quantified using LabWorks 4.0 software.Results: The quercetin-mediated inhibition of invasion was partially blocked by phorbol-12,13-dibutyrate (PDB), a PKC (protein kinase C) activator, and by doxorubicin, a PKC inhibitor. Only the proforms of MMP-9 (92 kDa) and MMP-2 (72 kDa) were detected by gelatin zymography. Quercetin dose-dependently decreased the gelatinolytic activity of pro-MMP-9. Doxorubicin also markedly reversed the quercetin-induced decrease. Quercetin showed a dose-dependent antagonism of increases in gelatinolytic activity of pro-MMP-9 induced by PDB and free fatty acid (another PKC activator).Conclusions: Together with the report that quercetin directly reduces PKC activity, the results reported here suggest that quercetin may inhibit the invasion of B16-BL6 cells by decreasing pro-MMP-9 via the PKC pathway. Published online: 30 October 2003     

The Inhibitory Effect of Endostatin and Doxycycline Administration on B16 Melanoma Angiogenesis and Cellular Proliferation

OBJECTIVE To investigate the effect of endostatin and doxycycline on melanoma cellular proliferation and tumor angiogenesis.METHODS The effects of endostatin and doxycycline were studied in mice transplanted with B16 melanoma cells.The mice were divided into 4 groups that were treated as follows:endostatin treatment(E group),doxycycline treatment(D group),endostatin plus doxycycline trearment(DE group),controls(C group) received no treatment.Following 9 days of treatment the tumor tissue was removed to compare the differences in the tumor necrotic rate and micro-vessel density(MVD) among the different groups.Immunohistochemical staining was conducted to detect the expression of proliferating cell nuclear antigen(PCNA)in the di?erent groups. RESULTS The MVD of the 3 experimental groups was significantly less than the control group,(F=10.888,P<0.05),indicating that doxycycline and endostatin can inhibit tumor angiogenesis by decreasing the tumor blood supply.This effect results in inhibition of tumor cellular proliferation and promotion of tumor cell necrosis.The tumor cell necrotic rate of the 3 experimental groups were all significantly higher than the C group(F=7.229,P<0.05) and the difference between the DE and C groups also was statistically significant.PCNA expression in all 3 experimental groups was statistically less than the C group(F=17.729,P<0.05). CONCLUSION The combined use of endostatin and doxycycline in vivo can influence PCNA expression and angiogenesis in melanoma,and significantly inhibit melanoma cellular proliferation.     

Matrix metalloproteinases 2 and 9 (gelatinases A and B) expression in malignant mesothelioma and benign pleura

Matrix metalloproteinases (MMPs), in particular the gelatinases (MMP-2 and -9), play a significant role in tumour invasion and angiogenesis. The expression and activities of MMPs have not been characterised in malignant mesothelioma (MM) tumour samples. In a prospective study, gelatinase activity was evaluated in homogenised supernatants of snap frozen MM (n=35), inflamed pleura (IP, n=12) and uninflammed pleura (UP, n=14) tissue specimens by semiquantitative gelatin zymography. Matrix metalloproteinases were correlated with clinicopathological factors and with survival using Kaplan-Meier and Cox proportional hazard models. In MM, pro- and active MMP-2 levels were significantly greater than for MMP-9 (P=0.006, P<0.001). Active MMP-2 was significantly greater in MM than in UP (P=0.04). MMP-2 activity was equivalent between IP and MM, but both pro- and active MMP-9 activities were greater in IP (P=0.02, P=0.009). While there were trends towards poor survival with increasing total and pro-MMP-2 activity (P=0.08) in univariate analysis, they were both independent poor prognostic factors in multivariate analysis in conjunction with weight loss (pro-MMP-2 P=0.03, total MMP-2 P=0.04). Total and pro-MMP-2 also contributed to the Cancer and Leukemia Group B prognostic groups. MMP-9 activities were not prognostic. Matrix metalloproteinases, and in particular MMP-2, the most abundant gelatinase, may play an important role in MM tumour growth and metastasis. Agents that reduce MMP synthesis and/or activity may have a role to play in the management of MM.     

膜型基质金属蛋白酶-1对乳腺癌细胞浸润能力的影响

目的 研究膜型基质金属蛋白酶-1（MT1-MMP）对乳腺癌细胞株浸润能力的影响,并初步探讨其作用机制。方法 用20μg／ml刀豆素（ConA）刺激乳腺癌细胞株MDA—MB-453,促使其表达MT1-MMP蛋白,并用免疫细胞化学和Western blot法检测;然后加入外源性MMP-2原酶（proMMP-2）,并用明胶酶谱分析法检测proMMP-2被激活的情况;最后用侵袭实验检测细胞株的浸润能力。实验中将细胞株分为4组：空白对照组、ConA组、MMP-2组和ConA＋MMP-2组,各组实验结果进行对比分析。结果 利用ConA刺激后,ConA组和ConA＋MMP-2组的细胞均表达MT1-MMP蛋白,另两组无MT1-MMP蛋白表达。明胶酶谱分析实验发现,MMP-2组只检测到72000原酶形式的MMP-2,ConA＋MMP．2组同时检测到72000原酶形式和64000活酶形式的MMP-2,其余两组检测不到任何形式MMP-2。侵袭实验结果显示,ConA＋MMP-2组细胞穿过Marigel胶的细胞数目明显多于其他组。结论 MT1-MMP能显著增强乳腺癌细胞株的浸润能力,其机制主要是通过激活MMP-2原酶,降解肿瘤周围的基质成分实现的。     

Differential Regulation of MMP-9 and TIMP-2 Expression in Malignant Melanoma Developed in Metallothionein/RET Transgenic Mice

We recently established a metallothionein-I(MT)/RET transgenic mouse line in which skin melanosis, benign melanocytic tumor and malignant melanoma develop stepwise. Malignant melanoma cells but not benign melanocytic tumor cells had metastatic ability in transgenic mice. In the present study, we investigated the expression of several matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) , including MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MT1– MMP, TIMP-1 and TIMP-2 , in these tumors. Western and northern blot analyses revealed that malignant transformation of melanocytic tumors developed in MT/RET transgenic mice accompanied with upregulation of MMP-9 and downregulation of TIMP-2. Expression of other MMP and TIMP genes examined was very low or undetectable in both benign and malignant tumors. Since activation of MMP-9 in malignant tumors was detected by gelatin zymography, these results suggest that imbalance of expression of the MMP-9 and TIMP-2 genes might be associated with metastatic ability of melanoma cells developed in MT/RET transgenic mice.     

Differential regulation of MMP-9 and TIMP-2 expression in malignant melanoma developed in metallothionein/RET transgenic mice.

We recently established a metallothionein-I(MT)/RET transgenic mouse line in which skin melanosis, benign melanocytic tumor and malignant melanoma develop stepwise. Malignant melanoma cells but not benign melanocytic tumor cells had metastatic ability in transgenic mice. In the present study, we investigated the expression of several matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs), including MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MT1-MMP, TIMP-1 and TIMP-2, in these tumors. Western and northern blot analyses revealed that malignant transformation of melanocytic tumors developed in MT/RET transgenic mice accompanied with upregulation of MMP-9 and downregulation of TIMP-2. Expression of other MMP and TIMP genes examined was very low or undetectable in both benign and malignant tumors. Since activation of MMP-9 in malignant tumors was detected by gelatin zymography, these results suggest that imbalance of expression of the MMP-9 and TIMP-2 genes might be associated with metastatic ability of melanoma cells developed in MT/RET transgenic mice.     

Increased expression of matrix metalloproteinases (MMP)-2, MMP-9, and the urokinase-type plasminogen activator is associated with progression from benign to advanced ovarian cancer.

Proteases are linked to the malignant phenotype of different solid tumors. Therefore, the expression of the matrix metalloproteinase (MMP)-2 and MMP-9 and of the serine protease urokinase-type plasminogen activator (uPA) and its inhibitor plasminogen activator inhibitor type 1 (PAI-1) in the progression of ovarian cancer was investigated. Gelatinolytic activity and protein expression of MMP-2 and MMP-9 were analyzed in tissue extracts of 19 cystadenomas and 18 low malignant potential (LMP) tumors, as well as 41 primary tumors of advanced ovarian cancer stage International Federation of Gynecology and Obstetrics IIIc/IV and their corresponding omentum metastases by quantitative gelatin zymography and Western blot. In the same tissue extracts, antigen levels of uPA and its inhibitor PAI-1 were determined by ELISA. Protein expression of pro-MMP-2 (72 kDa) and pro-MMP-9 (92 kDa as well as antigen levels of uPA and PAI-1 were low in benign ovarian tumors but increased significantly from LMP tumors to advanced ovarian cancers. The highest values of all of the proteolytic factors were detected in omentum metastases. Active MMP-2 enzyme (62 kDa) was detected only in ovarian cancer (66%) and corresponding metastases (93%) but never in benign or LMP tumors. The activation rate of MMP-2 to its active isoform was higher in the metastases. Comparing both proteolytic systems, higher PAI-1 concentrations were consistently found in cancers with high pro-MMP-9 expression. These data indicate that members of the plasminogen activator system, as well as the metalloproteinases MMP-2/9, increase with growing malignant potential of ovarian tumors. These findings are of particular relevance to the development of protease inhibitors as new therapeutic approaches in ovarian cancer.     

内皮抑素30肽对SGC-7901细胞增殖及转移能力的影响

目的研究经RGD修饰的人内皮抑素（endostatin）N-端的30肽对胃癌SGC-7901细胞增殖及转移能力的影响。方法人工合成人内皮抑素1～30位氨基酸(30肽,序列25～31由RGIRGAD改为RGDRGD)所对应的核苷酸序列,连接到质粒pTYB2中,再转化至大肠埃希菌BL21(DE3)中表达。用WST-1试剂检测30肽对SGC-7901细胞增殖的影响;细胞划痕实验和迁移实验观察30肽对细胞运动迁移的影响;明胶酶谱实验观察其对肿瘤转移相关基因MMP-2、MMP-9表达和分泌的影响。结果30肽能够有效抑制SGC-7901细胞的增殖、侵袭和迁移,同时还抑制了MMP-2和MMP-9的活性。结论30肽可能是通过下调MMP-2和MMP-9的活性而抑制SGC-7901细胞的转移,其在胃癌的临床治疗上具有潜在的应用前景。     

Enhanced production of matrix metalloproteinases and activation of matrix metalloproteinase 2 (gelatinase A) in human gastric carcinomas

We examined the production and tissue localization of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in gastric carcinoma tissues. MMP-I (tissue collagenase), MMP-9 (gelatinase B) and TIMP-2 were immunolocalized in carcinoma cells and MMP-2 (gelatinase A) on tumor cell membranes, whereas no or little immunostaining for MMP-3 (stromelysin-1) and TIMP-I was seen in carcinoma cells. Stromal cells in carcinoma tissue were also positively stained for these MMPs and TIMPs. MMP-2 immunostaining was observed exclusively on advanced gastric carcinoma cells and correlated with vascular invasion by tumor cells. Sandwich enzyme immunoassays revealed enhanced production of MMP-1, MMP-2, MMP-3, MMP-9 and TIMP-I by carcinoma tissues. Gelatinolytic activities were significantly higher in carcinoma samples than in normal controls. Using gelatin zymography, active forms of MMP-2 and MMP-9 were more frequently detected in carcinoma tissue, and the activation rate of the zymogen of MMP-2 (proMMP-2), but not that of proMMP-9, correlated well with degree of local invasion and lymphatic permeation. Our data indicate an enhanced production of 4 MMPs in gastric carcinoma tissue and suggest that activation of pro-MMP-2 may be a key step for spreading of gastric carcinoma cells. © 1996 Wiley-Liss, Inc.