基质金属蛋白酶-7和膜型基质金属蛋白酶-1在胃癌中的表达与胃癌临床病理特征的关系
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摘要
前言
侵袭和转移是恶性肿瘤主要生物学特性之一。原位恶性肿瘤在穿越自然屏障，侵袭周围组织并发生远处转移的过程中，细胞外基质和基底膜重塑是关键环节。细胞外基质的降解主要依靠各种蛋白水解酶的表达和激活。正常情况下，细胞外基质处于不断生长和不断降解的动态平衡中。在参与细胞外基质降解的酶系中，基质金属蛋白酶(MMPs)几乎能降解细胞外基质的所有成分。肿瘤细胞能分泌大量MMPs降解细胞外基质，而且转移倾向越高的肿瘤细胞分泌的MMPs含量越高。在肿瘤细胞介导的细胞外基质降解中MMPs起关键作用。因此，MMPs在肿瘤浸润和转移过程中的作用近年来备受关注。
基质金属蛋白酶-7(MMP-7)属于基质溶解素，由于缺乏与血红素结构蛋白同源序列而成为金属蛋白酶家族的最小成员。MMP-7的作用底物范围很广，主要是基质中的蛋白多糖和糖蛋白，如明胶、蛋白聚糖、弹性蛋白、纤维连接蛋白和酪蛋白。MMP-7由肿瘤细胞产生且能活化其它MMPs成员，如激活基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)等及灭活丝氨酸蛋白酶抑制剂。MMP-7的表达由癌基因、生长因子或激素调控。MMP-7在多种肿瘤中过表达，而且和肿瘤侵袭、转移、进展相关。
膜型基质金属蛋白酶-1(MT1-MMP)是MMPs家族中的一个亚家族。它是一种跨膜蛋白，存在于细胞膜上，具有降解细胞外基质的能力。它是明胶酶A的主要激活物，而明胶酶A是促进肿瘤侵袭和转移的主要基质金属蛋白酶之一。MT1-MMP在多种肿瘤组织中表达，与肿瘤的侵袭和转移相关。
目前的研究表明MMPs在多种肿瘤组织中过表达，在人类胃癌组织中MMPs的表达只有MMP-7和MT1-MMP几乎是选择性地定位于肿瘤细胞而无基质细胞着色。本研究应用免疫组织化学技术，同时检测MMP-7和MT1-MMP在胃癌组织和转移淋巴结中的表达，探讨它们与胃癌生物学行为的关系。
材料和方法
一、实验材料
1.实验标本随机选取中国医科大学附属第一临床医院肿瘤外科
1999年9月一2002年6月间手术切除的胃癌标本及其淋巴结60例。同时
选取非癌胃粘膜组织20例作为正常对照。
2.主要试剂鼠抗人MTI一MMP单克隆抗体和兔抗人MMP一7多克
隆抗体均购于福州迈新公司。sP免疫组化试剂盒和DAB显色剂均购于北
京中山公司。
3.主要仪器石蜡切片机;电热恒温干燥箱;光学显微镜。
二、实验方法
1.标本分组根据大体类型从标本中筛选出早期胃癌12例，Bor.
rm~1型胃癌12例，Bomnai双12型胃癌12例，Bornn田u6型胃癌12例，Bor-
rmann4型胃癌12例。每个大体类型为1组，共5组。
2.用免疫组织化学链霉素抗生物素蛋白一过氧化酶法(SP)染色检侧
MMP一7和MTI一MMP在胃癌中的表达。
3.结果判定标准以细胞浆有明显棕黄染色为阳性细胞。阴性(一):
无阳性细胞;弱阳性(+):阳性细胞数低于25%;阳性(++):阳性细胞数
25%一75%之间;强阳性(+++):阳性细胞数超过75%。勺
4.统计学处理用SPSS 11 .0统计分析软件对实验数据进行统计分
析，统计分析方法采用xZ检验和相关性分析，P<0.05有统计学意义。
实验结果
一、MMP一7和MTI一MMP在胃癌组织和淋巴结中的表达
MMP一7和MTI一MMP在非癌胃组织和无转移淋巴结中阴性表达，在
阳性表达的胃癌组织和转移淋巴结中多位于癌灶边缘，癌周基质阴性表达，
着色部位在胃癌细胞的胞浆中。60例胃癌组织中，MMP一7阳性率为
53.33%(32/60)，MTI一MMP阳性率为68.33%(41/60)。印例淋巴结中，
在无转移的28例淋巴结中MMP一7和MTI一MMP均为阴性表达。在有转
移的32例淋巴结中，MMP一7阳性率为75 .00%(24/32)，MTI一MMP阳性
率为84.38%(27乃2)。
二、MMP一7和MTI一MMP在胃癌组织中的表达与胃癌临床病理特征
的关系
MMP一7的阳性率在发生淋巴结转移的胃癌组织中为68.75%
(22/32)，在未发生淋巴结转移的胃癌组织中为35 .71%(10/28);淋巴结转
移距离:淋巴结未发生转移及淋巴结转移限于第I站(蕊N:)者为39 .47%
(巧乃8)，淋巴结转移至第n站和第n站以远(〕NZ)者为77.27%
(17/22);淋巴结转移数:0一4个者为47.50%(19/40)，5个以上者为65.
oo%(13/20);浸润深度:肿瘤侵袭至粘膜层、粘膜下层、肌层、浆膜下层(m
一Ss)者为32.00%(8/25)，肿瘤穿透浆膜层(Se)者为94.29%(33乃5);组
织学分型:肠型者为45.45%(10/22)，弥漫型者为57.89%(22乃8);生长
方式:团块状生长者为35 .71%(10/28)，巢状、弥漫状生长者为68.75%
(22/32);大体分型:局限型(B心 rr.1、2)为41 .67%(10/24)，浸润型(Borr.
3、4)为75·oo%(‘8/24一MMP一7的阳性率与淋巴结转移(犷二6.55，P<
0.05)、淋巴结转移距离(扩二8.00，P<0.01)、浸润深度(x，二23.35，P<
0.01)、生长方式(犷二6.55，p<0.05)、大体分型(xZ=5.49，p<0.05)相
关;与淋巴结转移个数(犷=1.64，P>0.05)和组织学分型(扩=0.87，P>
0.05)无相关性。
MTI一MMP的阳性率在发生淋巴结转移的胃癌组织中为90.63%(29/
32)，在未发生淋巴结转移的胃癌组织中为42.86%(12/28);淋巴结转移距
离:淋巴结未发生转移及淋巴结转移限于第I站(感Nl)者为5
Objective
Matrix metalloproteinases (MMPs) are well known proteinases that have been considered to play significant roles in tumor invasion and metastasis. This study examined the expression of MMP - 7 and MT1 - MMP in human gastric carcinoma and metastastic lymphonode by Immunohistochemical streptavidin -peroxidase (SP) method. Analyze the correlation between the expression of MMP - 7 and MT1 - MMP and tumor invasion, lymphonode metastasis and classification. Supply the theory basis for determination of gastric carcinoma invasion and metastasis. Direct the surgery and assistant treatment of gastric carcinoma.
Materials
1. Experimental specimens: Paraffin wax pieces were obtained at random from 60 patients with stomach neoplasm who underwent surgery in the Department of Oncology from September 1999 to June 2002, the Affiliated Hospital of China Medical University, Shenyang, China. After resection the samples were immediately frozen in 4% polydehyde. 20 cases of normal stomach tissues of the edge of tumor were selected at random as control.
2. Experimental reagents: rabbit anti human MMP - 7 polyclonal antibody and rat anti human MT1 - MMP monoclonal antibody ( neomarker company of Fuzhou) ; SP kit (zhongshan company of Beijing).
3. Experimental instruments: paraffin wax microtome, microscopy, drying machine.
Methods
The clinicopathologic findings were diagnosed by reviewing all hematoxylin and eosin (H&E) stained tissue sections.
From the specimens we screened 12 cases of early stage stomach neoplasm, 12 cases of Borrmannl, 12 cases of Bornnann2, 12 cases of BorrmannS, 12 cases of Borrmann4 according to the macroscopic type. Devide the specimens into 3 groups. Every kind of macroscopic type was as one group.
To determine the MMP - 7 and MT1 - MMP expression level of stomach neoplasm , Immunohistochemistry method was used.
The distribution of cells immunopositive for MMP - 7 and MT1 - MMP antibody was analyzed and taken a picture by microscopy.
Data were analyzed with SPSS 11. 0 statistic software. x1 test and linear regression was used. A P -value of <0.05 was considered significant.
Results
Immunohistochemical studies using the polyclonal MMP - 7 and MT1 -MMP antibody clearly showed positive staining in every grade stomach cells. The distribution of MMP - 7 and MT1 - MMP was at the cell membrane and cell plasm. There were no expression of MMP -7 and MT1 - MMP in normal gastric mucosa and lymphonode without metastasis.
The expression level of MMP - 7 and MT1 - MMP in gastric carcinoma correlated with lymphonode metastasis (P <0. 01) ,lymphonode metastasis distance ( P < 0.01) ,depth of invasion ( P < 0.05 ) ,macroscopic type ( P < 0.05 ), growth type ( P < 0. 01). The expression level of MT1 - MMP correlated with clinical histological classification and lymphonode metastasis number ( P < 0. 05 ). The expression level of MMP - 7 did not correlate with clinical histological classification and lymphonode metastasis number.
There was a significant positive correlation between expression of MMP - 7 in gastric carcinoma and lymphonode (r =0.955, P <0.01). There was a sig-
nificant positive correlation between expression of MTl - MMP in gastric carcinoma and lymphonode (r=0.927, P<0.05).
Tlie positive rate of MMP -7 in gastric carcinoma was 53.33% (32/60). The positive rate of MTl - MMP in gastric carcinoma was 68. 33% (41/60). There was a significant positive correlation between expression of MMP - 7 and MTl -MMP in gastric carcinoma (r=0.940, P<0.01).
Discussion
MMPs are related to invasion and metastasis of tumor. This study demonstrated gastric carcinoma cells express MMP - 7 at the border of focus mostly while normal gastric epithelial cell and matrix around of focus did not express MMP -7. The expression level of MMP -7 and MTl - MMP in gastric carcinoma correlated with lymphonode metastasis , lymphonode metastasis distance, depth of invasion ,macroscopic type ,growth type. The expression level of MTl -MMP correlated with clinical histological classification and
侵袭和转移是恶性肿瘤主要生物学特性之一。原位恶性肿瘤在穿越自然屏障，侵袭周围组织并发生远处转移的过程中，细胞外基质和基底膜重塑是关键环节。细胞外基质的降解主要依靠各种蛋白水解酶的表达和激活。正常情况下，细胞外基质处于不断生长和不断降解的动态平衡中。在参与细胞外基质降解的酶系中，基质金属蛋白酶(MMPs)几乎能降解细胞外基质的所有成分。肿瘤细胞能分泌大量MMPs降解细胞外基质，而且转移倾向越高的肿瘤细胞分泌的MMPs含量越高。在肿瘤细胞介导的细胞外基质降解中MMPs起关键作用。因此，MMPs在肿瘤浸润和转移过程中的作用近年来备受关注。
基质金属蛋白酶-7(MMP-7)属于基质溶解素，由于缺乏与血红素结构蛋白同源序列而成为金属蛋白酶家族的最小成员。MMP-7的作用底物范围很广，主要是基质中的蛋白多糖和糖蛋白，如明胶、蛋白聚糖、弹性蛋白、纤维连接蛋白和酪蛋白。MMP-7由肿瘤细胞产生且能活化其它MMPs成员，如激活基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)等及灭活丝氨酸蛋白酶抑制剂。MMP-7的表达由癌基因、生长因子或激素调控。MMP-7在多种肿瘤中过表达，而且和肿瘤侵袭、转移、进展相关。
膜型基质金属蛋白酶-1(MT1-MMP)是MMPs家族中的一个亚家族。它是一种跨膜蛋白，存在于细胞膜上，具有降解细胞外基质的能力。它是明胶酶A的主要激活物，而明胶酶A是促进肿瘤侵袭和转移的主要基质金属蛋白酶之一。MT1-MMP在多种肿瘤组织中表达，与肿瘤的侵袭和转移相关。
目前的研究表明MMPs在多种肿瘤组织中过表达，在人类胃癌组织中MMPs的表达只有MMP-7和MT1-MMP几乎是选择性地定位于肿瘤细胞而无基质细胞着色。本研究应用免疫组织化学技术，同时检测MMP-7和MT1-MMP在胃癌组织和转移淋巴结中的表达，探讨它们与胃癌生物学行为的关系。
材料和方法
一、实验材料
1.实验标本随机选取中国医科大学附属第一临床医院肿瘤外科
1999年9月一2002年6月间手术切除的胃癌标本及其淋巴结60例。同时
选取非癌胃粘膜组织20例作为正常对照。
2.主要试剂鼠抗人MTI一MMP单克隆抗体和兔抗人MMP一7多克
隆抗体均购于福州迈新公司。sP免疫组化试剂盒和DAB显色剂均购于北
京中山公司。
3.主要仪器石蜡切片机;电热恒温干燥箱;光学显微镜。
二、实验方法
1.标本分组根据大体类型从标本中筛选出早期胃癌12例，Bor.
rm~1型胃癌12例，Bomnai双12型胃癌12例，Bornn田u6型胃癌12例，Bor-
rmann4型胃癌12例。每个大体类型为1组，共5组。
2.用免疫组织化学链霉素抗生物素蛋白一过氧化酶法(SP)染色检侧
MMP一7和MTI一MMP在胃癌中的表达。
3.结果判定标准以细胞浆有明显棕黄染色为阳性细胞。阴性(一):
无阳性细胞;弱阳性(+):阳性细胞数低于25%;阳性(++):阳性细胞数
25%一75%之间;强阳性(+++):阳性细胞数超过75%。勺
4.统计学处理用SPSS 11 .0统计分析软件对实验数据进行统计分
析，统计分析方法采用xZ检验和相关性分析，P<0.05有统计学意义。
实验结果
一、MMP一7和MTI一MMP在胃癌组织和淋巴结中的表达
MMP一7和MTI一MMP在非癌胃组织和无转移淋巴结中阴性表达，在
阳性表达的胃癌组织和转移淋巴结中多位于癌灶边缘，癌周基质阴性表达，
着色部位在胃癌细胞的胞浆中。60例胃癌组织中，MMP一7阳性率为
53.33%(32/60)，MTI一MMP阳性率为68.33%(41/60)。印例淋巴结中，
在无转移的28例淋巴结中MMP一7和MTI一MMP均为阴性表达。在有转
移的32例淋巴结中，MMP一7阳性率为75 .00%(24/32)，MTI一MMP阳性
率为84.38%(27乃2)。
二、MMP一7和MTI一MMP在胃癌组织中的表达与胃癌临床病理特征
的关系
MMP一7的阳性率在发生淋巴结转移的胃癌组织中为68.75%
(22/32)，在未发生淋巴结转移的胃癌组织中为35 .71%(10/28);淋巴结转
移距离:淋巴结未发生转移及淋巴结转移限于第I站(蕊N:)者为39 .47%
(巧乃8)，淋巴结转移至第n站和第n站以远(〕NZ)者为77.27%
(17/22);淋巴结转移数:0一4个者为47.50%(19/40)，5个以上者为65.
oo%(13/20);浸润深度:肿瘤侵袭至粘膜层、粘膜下层、肌层、浆膜下层(m
一Ss)者为32.00%(8/25)，肿瘤穿透浆膜层(Se)者为94.29%(33乃5);组
织学分型:肠型者为45.45%(10/22)，弥漫型者为57.89%(22乃8);生长
方式:团块状生长者为35 .71%(10/28)，巢状、弥漫状生长者为68.75%
(22/32);大体分型:局限型(B心 rr.1、2)为41 .67%(10/24)，浸润型(Borr.
3、4)为75·oo%(‘8/24一MMP一7的阳性率与淋巴结转移(犷二6.55，P<
0.05)、淋巴结转移距离(扩二8.00，P<0.01)、浸润深度(x，二23.35，P<
0.01)、生长方式(犷二6.55，p<0.05)、大体分型(xZ=5.49，p<0.05)相
关;与淋巴结转移个数(犷=1.64，P>0.05)和组织学分型(扩=0.87，P>
0.05)无相关性。
MTI一MMP的阳性率在发生淋巴结转移的胃癌组织中为90.63%(29/
32)，在未发生淋巴结转移的胃癌组织中为42.86%(12/28);淋巴结转移距
离:淋巴结未发生转移及淋巴结转移限于第I站(感Nl)者为5
Objective
Matrix metalloproteinases (MMPs) are well known proteinases that have been considered to play significant roles in tumor invasion and metastasis. This study examined the expression of MMP - 7 and MT1 - MMP in human gastric carcinoma and metastastic lymphonode by Immunohistochemical streptavidin -peroxidase (SP) method. Analyze the correlation between the expression of MMP - 7 and MT1 - MMP and tumor invasion, lymphonode metastasis and classification. Supply the theory basis for determination of gastric carcinoma invasion and metastasis. Direct the surgery and assistant treatment of gastric carcinoma.
Materials
1. Experimental specimens: Paraffin wax pieces were obtained at random from 60 patients with stomach neoplasm who underwent surgery in the Department of Oncology from September 1999 to June 2002, the Affiliated Hospital of China Medical University, Shenyang, China. After resection the samples were immediately frozen in 4% polydehyde. 20 cases of normal stomach tissues of the edge of tumor were selected at random as control.
2. Experimental reagents: rabbit anti human MMP - 7 polyclonal antibody and rat anti human MT1 - MMP monoclonal antibody ( neomarker company of Fuzhou) ; SP kit (zhongshan company of Beijing).
3. Experimental instruments: paraffin wax microtome, microscopy, drying machine.
Methods
The clinicopathologic findings were diagnosed by reviewing all hematoxylin and eosin (H&E) stained tissue sections.
From the specimens we screened 12 cases of early stage stomach neoplasm, 12 cases of Borrmannl, 12 cases of Bornnann2, 12 cases of BorrmannS, 12 cases of Borrmann4 according to the macroscopic type. Devide the specimens into 3 groups. Every kind of macroscopic type was as one group.
To determine the MMP - 7 and MT1 - MMP expression level of stomach neoplasm , Immunohistochemistry method was used.
The distribution of cells immunopositive for MMP - 7 and MT1 - MMP antibody was analyzed and taken a picture by microscopy.
Data were analyzed with SPSS 11. 0 statistic software. x1 test and linear regression was used. A P -value of <0.05 was considered significant.
Results
Immunohistochemical studies using the polyclonal MMP - 7 and MT1 -MMP antibody clearly showed positive staining in every grade stomach cells. The distribution of MMP - 7 and MT1 - MMP was at the cell membrane and cell plasm. There were no expression of MMP -7 and MT1 - MMP in normal gastric mucosa and lymphonode without metastasis.
The expression level of MMP - 7 and MT1 - MMP in gastric carcinoma correlated with lymphonode metastasis (P <0. 01) ,lymphonode metastasis distance ( P < 0.01) ,depth of invasion ( P < 0.05 ) ,macroscopic type ( P < 0.05 ), growth type ( P < 0. 01). The expression level of MT1 - MMP correlated with clinical histological classification and lymphonode metastasis number ( P < 0. 05 ). The expression level of MMP - 7 did not correlate with clinical histological classification and lymphonode metastasis number.
There was a significant positive correlation between expression of MMP - 7 in gastric carcinoma and lymphonode (r =0.955, P <0.01). There was a sig-
nificant positive correlation between expression of MTl - MMP in gastric carcinoma and lymphonode (r=0.927, P<0.05).
Tlie positive rate of MMP -7 in gastric carcinoma was 53.33% (32/60). The positive rate of MTl - MMP in gastric carcinoma was 68. 33% (41/60). There was a significant positive correlation between expression of MMP - 7 and MTl -MMP in gastric carcinoma (r=0.940, P<0.01).
Discussion
MMPs are related to invasion and metastasis of tumor. This study demonstrated gastric carcinoma cells express MMP - 7 at the border of focus mostly while normal gastric epithelial cell and matrix around of focus did not express MMP -7. The expression level of MMP -7 and MTl - MMP in gastric carcinoma correlated with lymphonode metastasis , lymphonode metastasis distance, depth of invasion ,macroscopic type ,growth type. The expression level of MTl -MMP correlated with clinical histological classification and
引文
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7. Muller D, Breathach R, Engelmann A, Millon R, Bronner G, Flesch H, et al. Expression collagenase- related metalloproteinase genes in human lung or head and neck tumors. Int J Cancer, 1991,.48:550-6.
8. Basset P, Bellocq JP, Wolf C, Stoll I, Hutin P, Limacher JM, et al. A no-vel metalloproteinase gene specifically expressed in stromal cells of breast carcinomas. Nature, 1990, 348:699-704.
9. Pajouh MK, Bowden GT. Expression of metalloproteinase genes in human prostate cancer. J Cancer Res Clin Oncol, 1991, 117:144-50.
10. Rodgers WH, Osteen KG, Matrisian LM, Giudice LG, Gorstein F. Expres-sion and localization of matrilysin, matrix metalloproteinase -2, in human endometrium during the reproductive cycle. Am J Obstet Gymecol, 1993, 260:168-253.
11. Matri H-P, McNeil L, Thomas G, Davies M, Lovett DH. Molecular char-acterization of a low-molecularmass matrix metalloproteinase secreted by glomerular mesangial cells as PUMP -1. Biochem J, 1992, 285:899-905.
12. Pepper MS, Montesano R, Orel L, Vassalli J-D. Plasminogen activator inhibitor-1 is induced by a chondrocy TE-derived transforming growth factor-beta. Biochem Biophys Ret Commui, 1991,176:633-8.
13. Matrisian LM. The matrix-degrading metalloproteinases. Bioessays,1992,14:455-63.
14. Stilter-Stevenson WG, Liotta LA, Kleiner DE, Jr. Extracellur matrix:role of matrix metalloproteinase in tumor invasion and metastasis.FASEB J,1993,7:1434-41.
15. Sato H, Takino T, Okada Y, et al. A matrix metalloproteinase expressed on the surface of invasive tumor celles [J] Natuer, 1994, 370:61-65.
16. Moil M, Mimori K, Shiraishi T, et al. Analysis of MT1-MMP and MMP-2 expression in human gastric cancers[J]. Int J Cancer, 1997,74:316-21.
17. Okada A, Bellocq JP, Rouyer N, et al. Membrane -type matrix metallo-proteinase (MT-MMP) gene is expressed in stromal cells of human colon,breast, and head and neck carcinomas [J]. Proc Natl Acad Sci USA,1995, 92:2730-34.
18. Yamashita K, Azumano I, Okada Y, et al. Expression and tissue locaLiza-tion of matrix metalloproteinase-7 (matrilysin) in human gastric carcino-mas. ImpLications for vessel invasion and metastasis. Int J Cancer,1998,79:187-94.
19. Murphy G, Reynolds JJ, Hembry RM. Metalloproteinases and cancer inva-sion and metastasis[J]. Int J Cancer, 1989, 44:757-760.
20. Fang J, Shing Y, Wiederschain D, et al. Matrix metalloproteinase-2 is required for the switch to the angiogenic phenotype in a tumor mode[J].Proc Natl Acad Sci USA, 2000, 97:3884-3889.
21. McDonnell S, Navre M, Coffey RJ, et al. Expression and localization of the matrix metalloproteinase PUMP-1 (MMP-7) in human gastric and colon
carcinomas. Mol Carcinog, 1991,4:527-33.
22. Honda M, Mori M, Ueo H, et al. Matrix metalloproteinase -7 expression in gastric carcinomas. Gut, 1996, 39:444-8.
23．杜月光，陈慰浙，陈智，孔梅，万海同．人胃癌组织中基质金属蛋白酶-7的基因表达及意义[J]．中国癌症杂志，2003，13：201-3．
24. Senota A, Itoh F, Yamamoto H, et al. Relation of matrilysin messenger RNA expression with invasive activity in human gastric cancer. Clin Exp.Metastasis,1998, 16:313-21.
25. Nomura H, Sato H, Seiki M, et al. Expression of membrane-type matrix metalloproteinases in human gastric carcinomas [J]. Cancer Research,1995,55:3263-6.
26．万远廉，郭源，魏群等．膜型基质金属蛋白酶-1在人胃癌中的表达[J]．北京医科大学学报，2000，32(1)：61-3．
2. Douglas B. Invasion and metastasis. Cancer and Metastasis Rev,1996;15:77
3. Knox JD, Boreham DR, Walker JA, et al. Mapping of the metalloproteinase gene matrilysin (MMP-7) to human chromosomel 11 q21-q22.Cytogenet Cell Genet, 1996, 72:179-82.
4. Miyazaki K, Hattori Y, Umenishi F, Yasumitsu H, Umeda M. Purification and characterization of extracellular matrix-degrading metalloproteinase,matrin (Pump-1), secreted from human rectal carcinoma cell line. Cancer Res,1990, 50:7758-64.
5. Quantin B, Murphy G, Breathnach R. Pump-1 cDNA codes for a protein with characteristics similar to those of classical collagenase family members.Biochem J,1998,28:5327-33.
6. Parsons SL, Swatson SA, Brown PD, et al. Matrix metalloproteinases [J].Br J Surg, 1997, 84(2):160-6.
7. Muller D, Breathach R, Engelmann A, Millon R, Bronner G, Flesch H, et al. Expression collagenase- related metalloproteinase genes in human lung or head and neck tumors. Int J Cancer, 1991,.48:550-6.
8. Basset P, Bellocq JP, Wolf C, Stoll I, Hutin P, Limacher JM, et al. A no-vel metalloproteinase gene specifically expressed in stromal cells of breast carcinomas. Nature, 1990, 348:699-704.
9. Pajouh MK, Bowden GT. Expression of metalloproteinase genes in human prostate cancer. J Cancer Res Clin Oncol, 1991, 117:144-50.
10. Rodgers WH, Osteen KG, Matrisian LM, Giudice LG, Gorstein F. Expres-sion and localization of matrilysin, matrix metalloproteinase -2, in human endometrium during the reproductive cycle. Am J Obstet Gymecol, 1993, 260:168-253.
11. Matri H-P, McNeil L, Thomas G, Davies M, Lovett DH. Molecular char-acterization of a low-molecularmass matrix metalloproteinase secreted by glomerular mesangial cells as PUMP -1. Biochem J, 1992, 285:899-905.
12. Pepper MS, Montesano R, Orel L, Vassalli J-D. Plasminogen activator inhibitor-1 is induced by a chondrocy TE-derived transforming growth factor-beta. Biochem Biophys Ret Commui, 1991,176:633-8.
13. Matrisian LM. The matrix-degrading metalloproteinases. Bioessays,1992,14:455-63.
14. Stilter-Stevenson WG, Liotta LA, Kleiner DE, Jr. Extracellur matrix:role of matrix metalloproteinase in tumor invasion and metastasis.FASEB J,1993,7:1434-41.
15. Sato H, Takino T, Okada Y, et al. A matrix metalloproteinase expressed on the surface of invasive tumor celles [J] Natuer, 1994, 370:61-65.
16. Moil M, Mimori K, Shiraishi T, et al. Analysis of MT1-MMP and MMP-2 expression in human gastric cancers[J]. Int J Cancer, 1997,74:316-21.
17. Okada A, Bellocq JP, Rouyer N, et al. Membrane -type matrix metallo-proteinase (MT-MMP) gene is expressed in stromal cells of human colon,breast, and head and neck carcinomas [J]. Proc Natl Acad Sci USA,1995, 92:2730-34.
18. Yamashita K, Azumano I, Okada Y, et al. Expression and tissue locaLiza-tion of matrix metalloproteinase-7 (matrilysin) in human gastric carcino-mas. ImpLications for vessel invasion and metastasis. Int J Cancer,1998,79:187-94.
19. Murphy G, Reynolds JJ, Hembry RM. Metalloproteinases and cancer inva-sion and metastasis[J]. Int J Cancer, 1989, 44:757-760.
20. Fang J, Shing Y, Wiederschain D, et al. Matrix metalloproteinase-2 is required for the switch to the angiogenic phenotype in a tumor mode[J].Proc Natl Acad Sci USA, 2000, 97:3884-3889.
21. McDonnell S, Navre M, Coffey RJ, et al. Expression and localization of the matrix metalloproteinase PUMP-1 (MMP-7) in human gastric and colon
carcinomas. Mol Carcinog, 1991,4:527-33.
22. Honda M, Mori M, Ueo H, et al. Matrix metalloproteinase -7 expression in gastric carcinomas. Gut, 1996, 39:444-8.
23．杜月光，陈慰浙，陈智，孔梅，万海同．人胃癌组织中基质金属蛋白酶-7的基因表达及意义[J]．中国癌症杂志，2003，13：201-3．
24. Senota A, Itoh F, Yamamoto H, et al. Relation of matrilysin messenger RNA expression with invasive activity in human gastric cancer. Clin Exp.Metastasis,1998, 16:313-21.
25. Nomura H, Sato H, Seiki M, et al. Expression of membrane-type matrix metalloproteinases in human gastric carcinomas [J]. Cancer Research,1995,55:3263-6.
26．万远廉，郭源，魏群等．膜型基质金属蛋白酶-1在人胃癌中的表达[J]．北京医科大学学报，2000，32(1)：61-3．