MT1-MMP反义核酸抑制高转移人卵巢癌SW626细胞的侵袭效应

背景与目的:MT1-MMP(MMP-14)是新发现的一种膜型基质金属蛋白酶,研究表明MT1-MMP在肿瘤转移中发挥关键性作用。本研究应用反义核酸技术,观察了MT1-MMP反义核酸对高转移人卵巢癌SW626细胞体外生长和侵袭的抑制效应。方法:应用自行设计的引物,借助RT-PCR技术获得两端含不同限制酶位点的MT1-MMPcDNA片段,反向插入pcDNA3.1中,并应用脂质体介导的基因转染技术,将重组质粒导入SW626细胞中,应用MTT、Westernblot、明胶酶谱和体外侵袭实验等方法观察了SW626细胞转染前后,细胞生长、MT1-MMP蛋白表达、MMP-2和MMP-9酶活性及细胞体外侵袭能力等指标的变化。结果:成功构建了反义MT1-MMP真核表达载体(pMMP14as),将其转染入SW626细胞后,与空质粒转染组相比,MT1-MMP反义核酸转染组细胞MT1-MMP表达显著降低,抑制率为65.8%;MMP-2酶原的活化受到明显抑制;pMMP14as转染组的穿膜细胞百分数为(63.3±5.8)%,明显低于空质粒转染组(97.6±7.5)(P<0.05)%。结论:MT1-MMP反义核酸可明显抑制高转移SW626细胞体外生长和侵袭效应,提示MT1-MMP可作为人卵巢癌抗侵袭治疗的分子靶点。     

畸胎瘤细胞源性生长因子反义核酸载体对高度恶性人卵巢癌细胞增殖和侵袭的抑制效应及相关机制

目的 观察畸胎瘤细胞源性生长因子(PCDGF)反义核酸载体对高度恶性人卵巢癌细胞株Sw626和A2780增殖和侵袭的抑制效应,并初步探讨其相关机制.方法 采用二苯基溴化四氮唑蓝(MTT)法和Boyden小窒体外侵袭实验,检测PCDGF反义RNA真核表达载体对Sw626和A2780细胞增殖和侵袭能力的影响.采用Western blot技术,检测转染PCDGF反义RNA真核表达载体前后Sw626细胞cyclin D1和CDK4蛋自表达的变化.采用逆转录聚合酶链反应(RT-PCR)和明胶酶谱法,分析PCDGF反义RNA真核表达载体对Sw626细胞基质金属蛋白酶2(MMP-2)表达和活性的影响.结果 与空白对照组相比,PCDGF反义核酸载体转染组Sw626和A2780细胞的增殖抑制率分别为72.9%和70.9%,侵袭能力分别被抑制了62.9%和59.0%.转染组Sw626细胞cyclin D1和CDK4蛋白的表达水平分别为0.38±0.08和0.37±0.13,明显低于空白对照组(0.84±0.11和0.64±0.11,P<0.01).与空白对照组(0.89±0.09)相比,转染组Sw626细胞MMP-2 mRNA的表达水平(0.66±0.11)虽未见降低(P>0.05),但MMP-2酶原的活性被叨显抑制.结论 PCDGF反义核酸可显著抑制高度恶性人卵巢癌细胞株Sw626和A2780的增殖和侵袭能力,并逆转其部分恶性表型,这可能与其能下调cyclin D1和CDK4蛋白的表达并抑制MMP-2酶原的活性有关;PCDGF可以作为卵巢癌治疗的新靶点.     

寡核苷酸阻断ROCK-1蛋白对人卵巢癌细胞恶性行为的影响

目的通过反义寡核苷酸(ASODN)对ROCK-1的特异性阻断,探讨ROCK-1蛋白在卵巢癌转移中的作用。方法将ROCK-1反义寡核苷酸(ASODN)以脂质体介导,转染人卵巢癌细胞系SW626和Caov-3细胞,采用逆转录聚合酶链反应(RT-PCR)与Western blot印迹法,检测转染前后ROCK-1蛋白的表达,Boyden小室观察ROCK-1 ASODN对SW626和Caov-3细胞侵袭及迁移能力的影响,采用二苯基溴化四氮唑蓝(MTT)比色法,测定转染前后细胞增殖及黏附能力的变化。结果转染ROCK-1 ASODN后,2株细胞内ROCK-1蛋白的表达明显减少,最大抑制率可达49.0%;细胞的侵袭能力受到明显抑制,10μmol/L组和20μmol/L组SW626细胞的侵袭能力分别为对照组的75.6%±3.8%和54.7%±2.9%,Caov-3细胞为68.8%±4.7%和50.0%±4.5%;转染2种浓度ASODN的SW626和Caov-3细胞的随机运动能力分别为对照组的80.0%±1.3%、63.7%±1.9%、72.0%±1.3%和55.9%±2.5%;定向运动能力分别为对照组的83.9%±1.4%、64.1%±1.3%、72.5%±3.4%和54.5%±1.9%。转染后2株细胞的体外黏附能力和增殖能力,均未显示出明显的变化。结论ROCK-1蛋白的表达与人卵巢癌细胞的体外侵袭和迁移密切相关,阻断ROCK-1的表达,可有效地抑制卵巢癌肿瘤细胞的转移。     

Stroma-derived matrix metalloproteinase (MMP)-2 promotes membrane type 1-MMP-dependent tumor growth in mice

Matrix metalloproteinase-2 (MMP-2) is a stroma-derived MMP belonging to the type IV collagenase family. It is believed to mediate tumor cell behavior by degrading deposits of type IV collagen, a major component of the basement membrane. The membrane type 1-MMP (MT1-MMP) is a highly potent activator of MMP-2 and is expressed in many tumor and stromal cells. However, the roles played by stromal MMP-2 in tumor progression in vivo remain poorly understood. We established a colon epithelial cell line from an Mt1-mmp(-/-) mouse strain and transfected these cells with an inducible expression system for MT1-MMP (MT1rev cells). Following s.c. implantation into Mmp-2(+/+) mice and induction of MT1-MMP expression, MT1rev cells grew rapidly, whereas they grew very slowly in Mmp-2(-/-) mice, even in the presence of MT1-MMP. This MT1-MMP-dependent tumor growth of MT1rev cells was enhanced in Mmp-2(-/-) mice as long as MMP-2 was supplied via transfection or coimplantation of MMP-2-positive fibroblasts. MT1rev cells cultured in vitro in a three-dimensional collagen gel matrix also required the MT1-MMP/MMP-2 axis for rapid proliferation. MT1rev cells deposit type IV collagen primarily at the cell-collagen interface, and these deposits seem scarce at sites of invasion and proliferation. These data suggest that cooperation between stroma-derived MMP-2 and tumor-derived MT1-MMP may play a role in tumor invasion and proliferation via remodeling of the tumor-associated basement membrane. To our knowledge, this is the first study demonstrating that MT1-MMP-dependent tumor growth in vivo requires stromal-derived MMP-2. It also suggests that MMP-2 represents a potential target for tumor therapeutics.     

Sequence-specific silencing of MT1-MMP expression suppresses tumor cell migration and invasion: importance of MT1-MMP as a therapeutic target for invasive tumors

Membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14) has been believed a key enzyme in tumor invasion, because it is expressed in a variety of malignant human tumors, and overexpression of the enzyme enhances the ability of cellular invasiveness. However, it has not necessarily been clarified whether the endogenously expressed MT1-MMP in human tumors plays a critical role in their invasiveness. We used RNA silencing technology to downregulate the endogenous MT1-MMP expression in human tumor cells (fibrosarcoma HT1080 and gastric carcinoma MKN-28 cell lines), and evaluated the effect on the invasion of a reconstituted basement membrane (Matrigel). Transfection of a double-stranded RNA targeted to the MT1-MMP gene decreased the level of the enzyme to less than 10-20% without affecting production of other MMPs. According to the degree of silencing, activation of proMMP-2 was inhibited. CD44 shedding was also inhibited, but only in part. Decreased MT1-MMP levels were also reflected in reduced cell motility on hyaluronan (HA) and invasion in Matrigel. Thus, specific downregulation of MT1-MMP expression was sufficient to cause significant inhibition of the migration and invasion of tumor cells, even though other MMPs continued to be expressed.     

联合转染PTEN与PINCH siRNA对结直肠癌细胞增殖、侵袭及凋亡影响的机制研究

目的:研究联合转染PTEN与PINCH siRNA对结直肠癌SW620细胞增殖、侵袭和凋亡的影响。方法:在结直肠癌SW620细胞中转染pcDNA-PTEN或/和si-PINCH,qRT-PCR检测PINCH和PTEN mRNA的表达,Western blot检测PINCH、PTEN、CDK1、MMP-2、Bcl-2和 Bax蛋白表达, MTT法、Transwell实验和流式细胞术分别检测细胞增殖、侵袭能力和凋亡率。结果:在结直肠癌SW620细胞中联合转染pcDNA-PTEN和si-PINCH后,PTEN的mRNA和蛋白表达量显著升高（P＜0.05）,PINCH的mRNA和蛋白表达量显著降低（P＜0.05）;转染pcDNA-PTEN或/和si-PINCH均可抑制SW620细胞增殖和侵袭并促进细胞凋亡,抑制SW620细胞内增殖蛋白CDK1、侵袭蛋白MMP-2和抗凋亡蛋白Bcl-2的表达,促进凋亡蛋白Bax的表达;联合转染pcDNA-PTEN和si-PINCH比单独转染pcDNA-PTEN或si-PINCH效果更显著。结论:联合转染pcDNA-PTEN和si-PINCH可抑制结直肠癌SW620细胞增殖和侵袭并促进细胞凋亡,且比单独转染效果更显著。     

弗林蛋白酶抑制剂对乳腺癌MCF-7细胞生长和迁移的影响

目的 探讨乳腺癌转移的机制,为深入研究乳腺癌发生、发展机制提供理论基础.方法 应用不同浓度的弗林蛋白酶（Furin）抑制剂α1-PDX处理乳腺癌MCF-7细胞.用四甲基偶氮唑蓝（MTT）和克隆形成实验检测Furin抑制剂对MCF-7细胞增殖和克隆形成的影响.单层细胞迁移实验和Transwell实验检测MCF-7细胞迁移和浸润能力.Hoechst 33342/PI双染法检测细胞凋亡.酶联免疫吸附法检测细胞培养液中基质金属蛋白酶2（MMP-2）和MMP-9蛋白水平.Western blot检测细胞迁移相关蛋白MT1-MMP、血管内皮生长因子（VEGF）-C和VEGF-D水平.结果 不同浓度α1-PDX作用MCF-7细胞48 h以上时,细胞的生长受到抑制,集落形成降低,细胞凋亡率升高.但在低浓度情况下对细胞迁移和侵袭起抑制作用.α1-PDX降低了细胞内MT1-MMP、VEGF-C、VEGF-D的表达,同时细胞培养液上清中MMP-2和MMP-9浓度也低于对照组.结论 Furin抑制剂通过抑制乳腺癌MCF-7细胞MMP及VEGF表达抑制肿瘤的迁移能力.     

膜型基质金属蛋白酶-1在基质金属蛋白酶-2活化中的作用机制

目的:研究MMP-2的活化并探索其机制.方法:采用酶谱分析方法检测瘤细胞条件培养基中MMP-2在包被前后表达和活化的情况,用RT-PCR方法检测MT1-MMP mRNA的表达,观察MT1-MMP激活的MMP-2在细胞侵袭中的作用,Boyden小室膜侵袭实验检测细胞的侵袭能力.结果:条件培养基明胶酶谱分析结果显示仅在三维聚I型胶原包被组存在MMP-2的活性形式;培养于I型胶原组的人MCF-7细胞MT1-MMP mRNA表达量明显高于对照组,与对照组间差异显著(P<0.01).Boyden小室膜侵袭实验可见,用MT1-MMP的抗体处理组的细胞数明显少于未用MT1-MMP的抗体处理组,差异有统计学意义(P<0.01).结论:I型胶原成分可以通过上调MT1-MMP的水平来调控人乳腺癌MCF-7细胞MMP-2的活化,MT1-MMP激活的MMP-2可以增强人乳腺癌MCF-7细胞的侵袭能力.     

Expression and biological role of cytoglobin in human ovarian cancer

Loss of cytoglobin is found to be involved in the progression of several human cancers. However, its expression pattern and biological roles in human ovarian cancers are not clear. In this study, we examined cytoglobin expression in 118 archived ovarian cancer specimens using immunohistochemistry. A total of 72 specimens (61.0 %) showed cytoglobin downregulation. cytoglobin downregulation positively correlated with advanced FIGO stage and tumor grade. Cytoglobin plasmid transfection was performed in SKOV3 cell line and siRNA knockdown was carried out in SW626 cell line. MTT, colony formation assay and matrigel invasion assay were carried out to assess the role of cytoglobin on cell proliferation and invasion. Cytoglobin overexpression inhibited cell growth, invasion, cell cycle progression and cyclin D1 expression in SKOV3 cell line and its depletion promoted cell proliferation, invasion, cell cycle transition and cyclin D1 expression. In conclusion, cytoglobin is downregulated in ovarian cancers and associated with advanced stage. Our data provides evidence that cytoglobin regulates the ovarian cancer cell proliferation and invasion.     

The chemokine stromal cell derived factor-1 (CXCL12) promotes glioma invasiveness through MT2-matrix metalloproteinase

Chemokines have been found to alter tumor growth and metastasis. We have described previously that a particular chemokine receptor, CXCR4, was predominantly expressed on various glioma cell lines and in resected glioblastoma specimens. Herein, we have tested the ligand of CXCR4, stromal cell derived factor-1alpha (SDF-1alpha, CXCL12), on the response of human glioma cells. We found that SDF-1alpha increased the expression of membrane type-2 matrix metalloproteinase (MT2-MMP), but not the other MT-MMPs, MMP-2 or MMP-9. The SDF-1alpha enhanced MT2-MMP expression was blocked by a CXCR4 antagonist, AMD3100. Functional invasion assays showed that SDF-1alpha stimulated glioma cells to invade through matrigel-coated chambers and this effect was inhibited in glioma cells by the stable downregulation of MT2-MMP expression using small interfering RNA (siRNA). In vivo and at asymptomatic stages following intracerebral implant of cells, mice harboring MT2-MMP siRNA downregulated clones had smaller and less invasive tumors compared with mice implanted with non-specific siRNA control cells. Analyses at symptomatic stages demonstrate that mice with MT2-MMP siRNA clones survive longer than mice harboring control cells. These results highlight MT2-MMP as an effector of CXCR4 signaling in glioma cells, and they reveal the novel role of MT2-MMP in modulating tumor activity.     

Granulocyte-macrophage colony-stimulating factor upregulates matrix metalloproteinase-2 (MMP-2) and membrane type-1 MMP (MT1-MMP) in human head and neck cancer cells

Matrix metalloproteinase-2 (MMP-2) and membrane type 1-MMP (MT1-MMP) play an important role in the invasion and metastasis of head and neck squamous cell carcinoma (HNSCC), but the mechanism of their regulation is not clearly understood. Recently, granulocyte-macrophage colony-stimulating factor (GM-CSF) has been shown to be associated with cancer invasion and metastasis. We hypothesized that GM-CSF may upregulate MMP-2 and/or MT1-MMP expression in HNSCC cells, and may thereby influence their ability to invade and metastasize. We studied the effects of GM-CSF on the production of MMP-2 and MT1-MMP in HNSCC cell lines SAS and HSC-2. Gelatin zymography of conditioned media derived from HNSCC cells revealed a major band of 68 kDa, which was characterized as proMMP-2. GM-CSF stimulated the production of proMMP-2 in both cell lines in a dose-dependent manner. Treatment with 50 ng/ml GM-CSF for 24 h increased the proMMP-2 activity 3.4-fold in SAS cells and 2.3-fold in HSC-2 cells compared with untreated controls. Northern blot analyses demonstrated that GM-CSF led to elevated mRNA levels of MMP-2 and MT1-MMP in both cell lines. The results identify GM-CSF as a regulator of MMP-2 and MT1-MMP expression in certain types of HNSCC, and suggest that GM-CSF may contribute to the invasiveness of HNSCC through the regulation of MMP-2 and MT1-MMP expression.     

Membrane type 1 matrix metalloproteinase regulates collagen-dependent mitogen-activated protein/extracellular signal-related kinase activation and cell migration

Mitogen-activated protein kinase-extracellular signal-related kinase (ERK) kinase 1 (MEK1)/ERK signaling has been implicated in the regulation of tumor cell invasion and metastasis. Migration of HT1080 cells on type I collagen was suppressed by the matrix metalloproteinase (MMP) inhibitors BB94 and tissue inhibitor of metalloproteinase (TIMP)-2 but not by TIMP-1. TIMP-2-specific inhibition suggests that membrane type 1 MMP (MT1-MMP) is likely involved in this process. Activation of ERK was induced in HT1080 cells adhered on dishes coated with type I collagen, and this was inhibited by BB94. MMP-2 processing in HT1080 cells, which also was stimulated by cultivation on type I collagen, was inhibited by MEK inhibitor PD98059. Expression of a constitutively active form of MEK1 promoted MMP-2 processing concomitant with the increase of MT1-MMP levels, suggesting that MT1-MMP is regulated by MEK/ERK signaling. In addition, expression of the hemopexin-like domain of MT1-MMP in HT1080 cells interfered with MMP-2 processing, ERK activation, and cell migration, implying that the enzymatic activity of MT1-MMP is involved in collagen-induced ERK activation, which results in enhanced cell migration. Thus, adhesion of HT1080 cells to type I collagen induces MT1-MMP-dependent ERK activation, which in turn causes an increase in MT1-MMP levels and subsequent cell migration.     

Hypoxia stimulates breast carcinoma cell invasion through MT1-MMP and MMP-2 activation

The process of cancer cell invasion involves degradation of the extracellular matrix (ECM) by proteases, integrin adhesion and cell motility. The role of ECM degrading proteases on the hypoxia-induced invasion of breast carcinoma cells was investigated. Hypoxia markedly increased the invasion capacity of MDA-MB-231 and MDA-MB-435 breast carcinoma cell lines. Matrix metalloproteinase (MMP) inhibitors blocked the hypoxia-induced invasion, whereas other protease inhibitors had no effect. Antibodies or siRNAs blocking either membrane type-1 MMP (MT1-MMP) or MMP-2 were effective in reducing the hypoxia-induced invasion. Serum-free reconstitution experiments confirmed the involvement of the MT1-MMP/MMP-2/tissue inhibitor of metalloproteinase-2 complex in this hypoxia-induced response. Overexpression of MT1-MMP in a poorly invasive breast cancer cell line, T47-D, promoted hypoxia-induced invasion and MMP-2 activation. Cell surface accumulation and activation of MT1-MMP without apparent regulation at the mRNA or protein levels indicated a post-translational adaptive response to hypoxia. Inhibition of the small GTPase RhoA eliminated the hypoxia-induced invasion and blocked the localization of MT1-MMP to the plasma membrane. Zymographic and molecular analysis of human breast tumors showed a strong correlation between hypoxic microenvironments and MMP-2 activation without changes in MT1-MMP expression. Our studies suggest that hypoxic tumor microenvironments promote breast cancer invasion through an MT1-MMP-dependent mechanism.     

慢病毒介导的RNA干扰沉默KIF14表达对结肠癌细胞生长和转移的影响

目的:探讨慢病毒介导RNA干扰沉默KIF14表达对结肠癌SW480细胞生长和转移的影响。方法:采用脂质体法将构建的慢病毒靶向siRNA-KIF14干扰载体转入SW480细胞后,采用RT-PCR和Western blot检测KIF14 mRNA和蛋白的表达,MTT法、流式细胞仪和Transwell小室实验分别检测SW480细胞增殖、凋亡和侵袭,Western blot检测上皮-间质转化相关蛋白N-cadherin、E-cadherin和Vimentin的表达。结果:转染SW480细胞后成功下调KIF14 mRNA和蛋白的表达。与未转染细胞相比,下调KIF14的表达可抑制SW480细胞增殖、侵袭和上皮-间质转化,并促进细胞凋亡（P＜0.05）。结论:下调KIF14表达可抑制结肠癌SW480细胞生长和转移。     

Expression of E1AF, an ets-oncogene transcription factor, highly correlates with malignant phenotype of malignant melanoma through up-regulation of the membrane-type-1 matrix metalloproteinase gene

Matrix metalloproteinase (MMP) is closely involved in the degradation of extracellular matrix and confers invasive and metastatic potential to malignant tumors. MMP-2 is a type-IV collagenase secreted as a proenzyme that is activated on the surface of the tumor cell by membrane-type 1-MMP (MT1-MMP). MT1-MMP plays a critical role during tumor progression and metastasis. We investigated the expression levels of E1AF and MT1-MMP in malignant melanoma cell lines and specimens from patients in order to clarify the mechanisms responsible for the invasion and metastasis of malignant melanoma. High levels of E1AF and MT1-MMP mRNA expression were observed in melanoma cells by Northern blotting and real-time PCR. The expression level was highly correlated with an invasive potential determined by an in vitro invasion assay. The down-regulation of MT1-MMP was identified when E1AF was knocked down by RNA interference. These results suggest that E1AF plays a crucial role in the invasion and metastasis of malignant melanoma through up-regulating the MT1-MMP expression.     

Expression of membrane type 1 matrix metalloproteinase is associated with cervical carcinoma progression and invasion

Membrane type 1 matrix metalloproteinase (MT1-MMP) is frequently expressed by cancer cells and is believed to play an important role in cancer cell invasion and metastasis. However, little is known about the role of MT1-MMP in mediating invasiveness of cervical cancer cells. In this study, we examined MT1-MMP expression in 58 primary human cervical tissue specimens, including normal cervix, low-grade squamous intraepithelial lesions (LSIL), high-grade SILs (HSIL), and invasive carcinomas. We also evaluated MT1-MMP, MMP-2, and tissue inhibitor of metalloproteinase-2 expression in several cervical cancer-derived cell lines, human papillomavirus (HPV)-immortalized keratinocytes, and keratinocytes derived from a LSIL. Using in situ hybridization techniques to study the cervical tissue specimens, we found that MT1-MMP expression increases with cervical tumor progression (Spearman correlation coefficient = 0.66; P < 0.0001, exact test). Specifically, MT1-MMP expression is very low or absent in normal cervix and LSILs, is readily detectable in HSILs, and is very strongly expressed in nearly all invasive carcinomas. Most but not all cervical cancer-derived cell lines also expressed significant levels of MT1-MMP and MMP-2. Constitutive expression of exogenous MT1-MMP in cervical carcinoma-derived cells and HPV-immortalized keratinocytes with low endogenous levels of MT1-MMP induced invasiveness in collagen I, but this effect was not observed in LSIL-derived keratinocytes. Our results show that MT1-MMP is a key enzyme mediating cervical cancer progression. However, MT1-MMP alone is not always sufficient for inducing keratinocyte invasiveness at least in the collagen I invasion assay used in this study. Further studies of gene expression in preinvasive and invasive cervical cancers should assist with identification of additional critical factors mediating cervical cancer progression.     

Type 1 insulin-like growth factor regulates MT1-MMP synthesis and tumor invasion via PI 3-kinase/Akt signaling

The membrane type 1 matrix metalloproteinase (MT1-MMP) has been identified as a major activator of MMP-2 - a process involving the formation of a trimolecular complex with TIMP-2. We previously identified the IGF-I receptor as a positive regulator of MMP-2 synthesis. Here, we investigated the role of IGF-IR in the regulation of MT1-MMP. Highly invasive Lewis lung carcinoma subline H-59 cells express MT1-MMP and utilize it to activate their major extracellular matrix degrading proteinase-MMP-2. These cells were transiently transfected with a plasmid vector expressing a luciferase reporter gene downstream of the mouse MT1-MMP promoter. IGF-I treatment increased luciferase activity in the transfected cells by up to 10-fold and augmented endogenous MT1-MMP mRNA and protein synthesis by up to 2-3-fold, relative to controls. MT1-MMP induction and invasion were blocked by the PI 3-kinase inhibitors LY294002 and wortmannin and by rapamycin, but not by the MEK inhibitor PD98059. Overexpression of a dominant negative Akt mutant or of the tumor suppressor phosphatase and tensin homologue, PTEN, in these cells also caused a significant reduction in MT1-MMP expression and invasion. The results demonstrate that IGF-IR controls tumor cell invasion by coordinately regulating MMP-2 expression and its MT1-MMP-mediated activation and identify PI 3-kinase/Akt/mTOR signaling as critical to this regulation.     

Effects of E-cadherin transfection on gene expression of a gallbladder carcinoma cell line: repression of MTS1/S100A4 gene expression

E-cadherin is important in cell-to-cell adhesion and controls cell polarity and tissue morphology. Loss of E-cadherin expression occurs in various human tumors and is the first step in cancer invasion and metastasis. We demonstrate that the exogenous expression of E-cadherin transfected into G-415 GB cells not only increases cell-to-cell adhesion but also reduces in vitro cell proliferation, motility and invasion. Our aim was to determine what genes are most affected by the exogenous expression of E-cadherin in GB cancer cells. We analyzed gene expression pertaining to cell proliferation, motility and invasion. Conventional RT-PCR was performed for these genes; quantitative RT-PCR was carried out on genes exhibiting altered expression. Conventional RT-PCR revealed that E-cadherin transfection suppressed expression of mts1 mRNA and increased that of c-myc and MT1-MMP. In quantitative RT-PCR analysis, levels of c-myc and MT1-MMP mRNA were elevated by to 2.56- and 2.22-fold, respectively, in the E-cadherin transfectant, whereas mts-1 was 7.14-fold suppressed compared to parental cells. These results indicated that expression of mts1 mRNA was most affected by E-cadherin transfection. Immunocytochemical analysis of transfectant and parental cells demonstrated an inverse correlation in E-cadherin and mts1 expression. Immunohistochemical analysis of 37 GB cancer specimens confirmed this observation in vivo. Loss of E-cadherin expression followed by expression of the mts1 gene may be an important event for increasing cell proliferation, motility and invasion activity in the progression of GB cancer.