The association of transforming growth factor-beta 1 with myometrial invasion of endometrial carcinomas through effects on matrix metalloproteinase

OBJECTIVE: The association of transforming growth factor-beta 1 (TGF-beta 1) with a matrix metalloproteinase (MMP) and a tissue inhibitor of metalloproteinase (TIMP), as well as myometrial invasion of endometrial cancer was studied. METHODS: The effects of TGF-beta 1 on cellular invasiveness, gelatinase activity, and expression of TIMP-1 were examined in 2 endometrial adenocarcinoma cell lines, KLE and Ishikawa. Plasma was obtained from 8 endometrial cancer patients with Stage-Ia disease, from 6 with Stage-Ib disease, and from 4 with Stage-Ic disease, and the levels of TGF-beta 1 were measured by enzyme immunoassays. The immunohistochemical expression of MMP-9, TIMP-1, TGF-beta 1, and TGF-beta receptor Type I in tumor tissue from the same patients also was detected. RESULTS: Invasiveness, gelatinase activity, and the expression of TIMP-1 were higher in KLE cells than in Ishikawa cells, and they were increased by treatment with rTGF-beta 1. The expression of TGF-beta receptor Type I was higher in KLE cells than in Ishikawa cells, which were unresponsive to exogenous TGF-beta 1. The plasma levels of TGF-beta 1 were greater in Stage-Ib and Stage-Ic patients than in Stage-Ia patients. MMP-9 and TGF-beta receptor Type I were expressed mainly in tumor cells, while TIMP-1 and TGF-beta 1 were localized in both tumor epithelial cells and stromal cells. MMP-9 and TIMP-1 were expressed only in Stage-Ib and Stage-Ic patients, although TGF-beta 1 and TGF-beta receptor Type I were ubiquitous. CONCLUSIONS: Myometrial invasion of endometrial cancers involves an increase in gelatinase activity, regulated to some extent by TGF-beta 1 in an autocrine or paracrine fashion.     

基质金属蛋白酶及其组织抑制因子在子宫内膜癌中的表达及其意义

目的　研究基质金属蛋白酶 (matrixmetalloproteinases,MMPs) 2、9及其组织抑制因子(tissueinhibitorofmetalloproteinases ,TIMPs) 1、2在子宫内膜癌中的表达 ,探讨其与子宫内膜癌浸润转移的关系。方法　应用链霉菌抗生物素蛋白 过氧化物酶免疫组织化学方法和明胶酶谱法对 37例内膜癌及 7例绝经期妇女子宫内膜组织中MMP 2、MMP 9、TIMP 1、TIMP 2蛋白及其活性进行检测。结果　MMP 2、MMP 9及TIMP 1、TIMP 2蛋白主要分布在内膜癌细胞、血管内皮细胞及绝经期子宫内膜腺上皮细胞中 ,在间质细胞中也有少量表达。内膜癌细胞中 ,MMP 2、MMP 9及TIMP 1蛋白的表达 ,病理分级为G3内膜癌的强阳性率分别为 73%、2 0 %及 6 7% ,高于G2 (13%、0及 2 7% )、G1 者 (均为 0 ,P<0 0 5 ) ;深肌层浸润内膜癌的强阳性率分别为 6 3%、16 %及 6 8% ,高于浅肌层浸润的 8%、0及 0 (P<0 0 1) ;有淋巴结转移者的强阳性率分别为 4例中 4例、4例中 3例及 4例中 4例 ,高于无淋巴结转移者的 2 5 %、0及 2 5 % (P <0 0 5 ) ;手术病理分期为Ⅲ～Ⅳ期者强阳性率分别为 5例中 5例、5例中 3例及 5例中 5例 ,高于Ⅰ～Ⅱ期者的 30 %、0及 30 % (P <0 0 5 ) ;TIMP 2蛋白在不同病理分级、肌层浸润、淋巴结转移和手术病理分期的内膜癌细     

Activity of matrix metalloproteinases -2 and -9 (MMP-2 and MMP-9) and content of their tissue inhibitors in endometrial cancer--a preliminary study

OBJECTIVES: Matrix metalloproteinases-2 and -9 (MMP-2 and MMP-9) are enzymes degrading collagen type IV and other components of the basement membrane. Their activity is suppressed by tissue inhibitors of metalloproteinases--TIMP-1 and TIMP-2. Substantial evidence indicates that MMP2 and MMP-9 play an important role in the spread of malignant tumours. The aim of the study was to evaluate the activity of MMP-2 and MMP-9 and contents of their inhibitors: TIMP-1 and TIMP-2 in endometrial cancer and normal endometrium. MATERIAL AND METHODS: Material for the study comprised 28 samples of endometrial cancers and 15 samples of normal endmetrium. A two-step method for extraction of MMPs was applied. The activity of MMP-2 and MMP-9 was measured with semi-quantitative zymography. TIMP-1 and TIMP-2 contents were measured with ELISA method. RESULTS: Mean activity and activation ratio of MMP-9 was significantly higher in endometrial cancers compared with normal myometrium, whereas mean activity and activation ratio of MMP-2 did not differ significantly between investigated groups. Mean content of TIMP-1 and TIMP-2 did not differ between cancer and control tissues. No unequivocal association between activity of investigated MMPs or contents of their inhibitors and clinicopathological features of endometrial cancers was observed. CONCLUSIONS: Results of the study suggest that MMP-9 may play an important role in the progression of endometrial cancer, whereas MMP-2 does not seem to be involved in this process. Action of MMP-9 may be further enhanced by relative deficiency of TIMP-1.     

基质金属蛋白酶-2及9与子宫内膜腺癌临床及预后的关系

目的探讨基质金属蛋白酶-2(MMP-2)和9(MMP-9)在正常子宫内膜组织、子宫内膜腺癌组织中的表达,及其与子宫内膜腺癌临床和预后的关系。方法2005年10月至2009年3月在中山大学附属中山医院采用免疫组织化学EnVision法,检测100例子宫内膜腺癌组织中MMP-2、MMP-9蛋白的表达,以30例正常子宫内膜组织作为对照组。结果MMP-2、MMP-9在正常子宫内膜增生期表达增加而在早中分泌期几乎无表达(P>0.05)。MMP-2、MMP-9蛋白在子宫内膜腺癌组织中的表达为74.0%和78.0%,显著高于正常子宫内膜组织(50.0%、56.7%)(P<0.05);随着子宫内膜腺癌临床分期、病理组织学分级的提高,MMP-2、MMP-9的表达也随之增强,且有统计学意义(P<0.05);在伴有淋巴结转移的子宫内膜腺癌组织中,两者表达(90.0%、60.0%)高于无淋巴结转移组(28.6%、23.8%)(P<0.05)。子宫内膜组织中MMP-2与MMP-9蛋白的表达呈正相关(r=0.545,P<0.001)。MMP-2和MMP-9的表达与患者生存率相关(P<0.05)。结论MMP-2、MMP-9在子宫内膜腺癌发...     

雌激素受体亚型α对子宫内膜癌细胞HEC-1B侵袭能力调控的研究

目的：特异性下调子宫内膜癌细胞中的ERα基因，探讨E胁亚型表达在子宫内膜癌侵袭中的作用。方法：将ERa的小干扰RNA（siRNA-small interfering RNA）转染子宫内膜癌细胞HEC-1B，通过RT-PCR和Western blot证实ERα基因的有效阻断。通过transwell小室法检测下调ERa基因表达前后HEC-1B细胞的侵袭能力；应用RT-PCR检测转染前后细胞MMP-2、MMP-9、TIMP-1和TIMP-2 mRNA表达水平的变化；Western blot及明胶酶谱分别检测细胞分泌TIMP-1、TIMP-2、MMP-2和MMP-9蛋白的水平。结果：（1）将ERα-siRNA转染HEC-1B细胞后，转染效率大于90％，ERα mRNA及蛋白的表达水平均明显下调（分别为72％，67％）；（2）下调ERα基因表达后，肿瘤细胞的侵袭能力下降（P〈0．05）；在mRNA水平和蛋白水平均可检测到ERα-siRNA组细胞的MMP-2、MMP-9表达下降（P〈0．05），TIMP-1、TIMP-2表达增加（P〈0．05）。结论：使用ERα-siRNA能够有效地阻断ERa基因表达；子宫内膜癌细胞中，17β-雌二醇对MMPs／TIMPs具有调节作用，这种作用可通过ERα介导；ERα表达水平影响子宫内膜癌细胞的侵袭能力。     

Endometrial tumor invasiveness is related to metalloproteinase 2 and tissue inhibitor of metalloproteinase 2 expressions

Matrix metalloproteinase (MMPs) expression has been linked to gynecological tumor aggressiveness. The objective of this study was to determine MMP-2, MMP-7, and MMP-9 and tissue inhibitors of metalloproteinases (TIMP)-1 and TIMP-2 expression in endometrial malignancies and their relation to clinical and histologic parameters. Formalin-fixed, paraffin-embedded tumor samples from 50 patients with endometrial carcinoma treated between 1999 and 2004 were stained with specific monoclonal antibodies. The tumors were grouped according to the FIGO classification. The staining results were compared to histologic and clinical data. Semiquantitative analysis of MMP and TIMP expression showed a significant difference in TIMP-2 expression according to the histologic subtype (P = 0.03) and also a trend towards a difference in MMP-9 expression (P = 0.05). MMP-2 expression increased and TIMP-2 expression fell as the histologic grade increased (P = 0.0007, P < 0.0001, respectively). MMP-2 expression correlated with lymph node metastasis (P = 0.04), while TIMP-2 expression correlated with the depth of myometrial invasion (P = 0.01), vasculolymphatic space involvement (P = 0.02), and lymph node metastasis (P = 0.0003). These results support the involvement of MMPs and TIMPs in endometrial tumor growth and progression. High MMP-2 and low TIMP-2 expression were the most potent markers of endometrial tumors with a high risk of local and distant spread.     

Functional role of matrix metalloproteinases in ovarian tumor cell plasticity

OBJECTIVE: We previously demonstrated that aggressive ovarian cancer cells are able to display in vitro vasculogenic mimicry, which is reflected by their ability to form vasculogenic-like networks in 3-dimensional cultures and to express vascular cell-associated markers. The goal of this study was to examine the functional role of specific matrix metalloproteinases in the formation of vasculogenic-like networks and extracellular matrix remodeling in vitro. We also investigated the clinical relevance of matrix metalloproteinase-2 and -9 and membrane type 1-matrix metalloproteinase in human ovarian cancers with evidence of tumor cell-lined vasculature. STUDY DESIGN: Ovarian cancer cells (A2780-PAR, SKOV3, and EG) were seeded onto separate 3-dimensional cultures that contained either Matrigel or type I collagen, in the absence of endothelial cells or fibroblasts. These cultures were treated with either chemically modified tetracycline-3 (general matrix metalloproteinase inhibitor), recombinant tissue inhibitor of metalloproteinase-1 or -2, or function-blocking antibodies to matrix metalloproteinase-2 or -9 or membrane type 1-matrix metalloproteinase. In addition, 78 invasive epithelial ovarian cancers were evaluated for expression of matrix metalloproteinase-2 and -9 and membrane type 1-matrix metalloproteinase and correlated with various clinical parameters. RESULTS: The aggressive ovarian cancer cells (SKOV3 and EG) were able to form in vitro vasculogenic-like networks and contract 3-dimensional collagen I gels, whereas the poorly aggressive A2780-PAR cell line did not. Chemically modified tetracycline-3 completely blocked the network formation. Blocking antibodies to matrix metalloproteinase-2 and membrane type 1-matrix metalloproteinase inhibited the formation of the vasculogenic-like networks and collagen gel contraction, but the antibody to matrix metalloproteinase-9 had no effect on network formation and minimal effect on gel contraction. Treatment of 3-dimensional cultures with tissue inhibitor of metalloproteinase-2 retarded the network formation and only small, partially developed structures were noted that did not form network connections. Tissue inhibitor of metalloproteinase-1 had no appreciable effect on the extent or efficiency of network formation. Human invasive ovarian cancers with evidence of tumor cell-lined vasculature were significantly more likely to have strong epithelial and stromal matrix metalloproteinase-2 and -9 and membrane type 1-matrix metalloproteinase expression (all probability values were <.05). CONCLUSION: Matrix metalloproteinase-2 and membrane type 1-matrix metalloproteinase appear to play a key role in the development of vasculogenic-like networks and matrix remodeling by aggressive ovarian cancer cells. Human ovarian cancers with matrix metalloproteinase overexpression are more likely to have tumor cell-lined vasculature. These results may offer new insights for consideration in ovarian cancer treatment strategies.     

Regulation of matrix metalloproteinases and their inhibitors in uterine endometrial cells of patients with and without endometriosis

OBJECTIVE: To determine whether alterations in the secretion and regulation of matrix metalloproteinases (MMPs) and their inhibitors are present in uterine endometrial cells from endometriosis patients. STUDY DESIGN: In an in vitro study, uterine endometrial cells from 19 regularly cycling women with and 32 without endometriosis were treated with diethyl stilbestrol, promegestone (R5020), interleukin-1 (IL-1) and tumor necrosis factor a (TNF-alpha). Culture supernatants were assayed for MMPs 1, 2, 3, and 9, and for tissue inhibitors of MMP (TIMP-1 and TIMP-2) by ELISA. RESULTS: MMP-3 was secreted in high concentrations, moderate concentrations were seen for MMP-1 and MMP-2, and very low concentrations for MMP-9. Substantially more TIMP-1 than TIMP-2 was secreted. MMP-1 and MMP-3 were uniformly attenuated by R5020, while MMP-2 was not influenced by hormone treatment. MMP-3 was upregulated by TNF-alpha in all samples while IL-1 only increased secretion in cells from endometriosis patients. CONCLUSION: The upregulation of MMP-3 by IL-1 may contribute to an increased invasiveness of uterine endometrial fragments in endometriosis patients.     

Gelatinase A-immunoreactive protein in ovarian lesions- prognostic value in epithelial ovarian cancer.

OBJECTIVE: Gelatinase A (MMP-2) is a member of the matrix metalloproteinase family and it degrades the major component of the basement membrane, type IV collagen. MMP-2 has been linked to invasion in different types of cancer. METHOD: We have studied the localization of MMP-2 in 18 benign, 3 borderline, and 33 malignant ovarian lesions by immunohistochemical stainings using a monoclonal antibody against MMP-2. RESULTS: MMP-2-immunoreactive protein was localized in epithelial cells and in fibroblasts. Two types of cytoplasmic staining were observed, a diffuse and a granular pattern. The diffuse staining model was found more often. In 19% of the cases, both staining patterns were present in epithelial cells. Granular staining was found in epithelial cells in cystadenomas and in ovarian cancer cells. The pattern of MMP-2 positivity in fibroblasts was diffuse. MMP-2 positivity in cancer cells was associated with recurrent disease (P < 0.05) in ovarian cancers. MMP-2 negativity in fibroblasts correlated to Grade 3 (P < 0.01), Stage III-IV (P < 0.001), recurrency (P < 0.05), and refractory disease (P < 0.05) in ovarian cancer. The relative survival rate was 32% in patients with an MMP-2-positive ovarian cancer, 57% in patients with an MMP-2-negative ovarian cancer, and 19% in patients with MMP-2 positivity in cancer cells and concomitant negativity in stromal fibroblasts. The disease-free 5-year survival rates were 25, 57, and 12.5%, respectively. CONCLUSIONS: These data suggest that MMP-2 may contribute to poor prognosis of ovarian cancer.     

MMP-1 and MMP-2 in the cervix uteri in different steps of malignant transformation--an immunohistochemical study.

OBJECTIVES: Enzymatic degradation of the extracellular matrix (ECM) represents a key element in the multistage process of tumor invasion and metastasis. This process requires extensive degradation of ECM components such as basement membrane collagen (type IV) and interstitial collagen (type I, II, III). Matrix metalloproteinase-2 (MMP-2) specifically cleaves collagen type IV, the major collagen of the basement membrane. MMP-1 digests interstitial collagen type I and III, the main collagen types of the stromal extracellular matrix. We investigated protein levels of MMP-1 and MMP-2 in different stages of malignant transformation. METHODS: Using the APAAP method we analyzed 10 normal cervical tissues, 11 cervical intraepithelial neoplasia 1 (CIN 1), 8 CIN 2 and 10 CIN 3 lesions, and 15 invasive squamous cell carcinomas. These data were compared with the HPV DNA status tested by hybrid capture II. RESULTS: Only a few isolated epithelial cells stained positively for MMP-1 and MMP-2 in normal cervical tissue and CIN 1 lesions. The CIN 2 and CIN 3 group displayed a heterogeneous distribution of MMP expression. 3 CIN 2 and 8 CIN 3 lesions showed strong MMP-2 and weak MMP-1 expression in the dysplastic epithelial cells. 5 CIN 2 and 2 CIN 3 lesions stained negatively. Invasive carcinomas showed a coexpression for MMP-1 and MMP-2 in malignant epithelial cells and peritumoral stroma cells. All MMP-2-positive cases tested positive for the HPV high-risk group. CONCLUSIONS: The expression of MMP-2 protein in preinvasive lesions of the cervix uteri and a consecutive coexpression of MMP-1 and MMP-2 in invasive cancer suggest a gradually increasing invasive potential. MMP-2 expression, when focally observed in high-grade squamous intraepithelial lesions of the cervix, may indicate tumor areas with an increased risk for invasive growth.     

Expression of matrix metalloproteinase-26 and tissue inhibitor of matrix metalloproteinase-3 and -4 in endometrium throughout the normal menstrual cycle and alteration in users of levonorgestrel implants who experience irregular uterine bleeding

OBJECTIVE: To determine the expression of matrix metalloproteinase (MMP-26) and tissue inhibitor of MMP (TIMP) in the endometrium of women with normal menstrual cycles compared with users of levonorgestrel implants. DESIGN: Prospective observational study. SETTING: Academic research center. PATIENT(S): Fifty patients with normal menstrual cycles who requested permanent surgical sterilization (tubal ligation) and 35 users of levonorgestrel implants. INTERVENTION(S): Endometrial biopsy. MAIN OUTCOME MEASURE(S): Expression of MMP-26, TIMP-3, and TIMP-4 by immunohistochemistry and semiquantitative analysis of staining intensity by using the H score. RESULT(S): Endometrium from women with a normal menstrual cycle and users of levonorgestrel implants expresses MMP-26, TIMP-3, and TIMP-4. These substances are present in various types of endometrial cells; expression is strongest in surface and glandular epithelial cells, followed by vascular endothelial and endometrial stromal cells. Inflammatory and immune-related cells also stained strongly for MMP-26 and TIMPs. Semiquantitative analysis of the staining intensity of endometrial epithelial and stromal cells indicated that expression of MMP-26, TIMP-3, and TIMP-4 peaks during the early to mid-luteal phase. Expression of MMP-26 is elevated in users of levonorgestrel implants who experienced irregular uterine bleeding. CONCLUSION(S): Endometrial expression of MMP-26 and TIMP-4 is present throughout the menstrual cycle and is elevated during the early to mid-luteal phase in normally cycling women. Further elevations in MMP-26 are seen in users of levonorgestrel implants who experience irregular uterine bleeding. These substances thus seem to play a role in hormonal regulation and endometrial tissue remodeling.     

Invasiveness corresponds to differentiation rather than to proteinase secretion in endometrial cancer cell lines

Local invasiveness is an important prognostic factor in endometrial carcinoma. To study the role of two groups of secreted proteinases (serine proteinases and matrix metalloproteinases) in this process, we examined three endometrial cancer cell lines (Ishikawa HEC 1A, AN3CA) for their invasiveness in vitro. Additionally, we considered the secretion of urokinase type plasminogen activator (uPA), plasminogen activator inhibitor 1 and 2 (PAI-1 and PAI-2), as well as matrix metalloproteinases (MMP) 1, 2, 3, and 9, and their inhibitors TIMP-1 and TIMP-2. Compared to the highly invasive fibrosarcoma cell line HT 1080, Ishikawa displayed low and AN3CA moderate invasiveness, while HEC 1A cells were almost as invasive as HT 1080 cells. Ishikawa cells secreted the highest amounts of proteinases. Cytokine and steroid treatments upregulated MMP-1 in all cell lines while the effects were heterogeneous regarding other proteinases and inhibitors. No effect of these treatments on invasiveness could be detected. Both basal secretion and regulation of the proteinases tested in this set of experiments seem to be markers of differentiation rather than of invasiveness.     

Endometrial bleeding in hormone replacement therapy users: preliminary findings regarding the role of matrix metalloproteinase 9 (MMP-9) and tissue inhibitors of MMPs

OBJECTIVE: To establish the effect of hormone replacement therapy (HRT) on the expression of matrix metalloproteinase 9 (MMP-9) and the tissue inhibitor of MMPs, TIMP-1, in the endometrium of postmenopausal and perimenopausal women. DESIGN: Prospective observational study. SETTING: United Kingdom teaching hospital. PATIENT(S): Thirty-one perimenopausal and postmenopausal HRT recipients, with a control group of eight postmenopausal women not undergoing HRT. INTERVENTION(S): Prospective record of bleeding patterns and endometrial biopsy. MAIN OUTCOME MEASURE(S): Endometrial histology, bleeding patterns, MMP-9, and TIMP-1 expression. RESULT(S): MMP-9 and TIMP-1 are expressed in benign postmenopausal endometrium. Expression of both molecules is reduced in HRT recipients compared with non-HRT recipients. CONCLUSION(S): Exposure to HRT appears to alter endometrial expression of MMP-9 and TIMP-1 and also the local balance between these molecules. This alteration may promote breakdown of the endometrial extracellular matrix and blood vessels and hence bleeding.     

MMP-2/TIMP-2在人及裸鼠异位子宫内膜组织中的表达及与雌、孕激素的关系

目的:探讨MMP-2/TIMP-2在子宫内膜异位症发病机制中的作用及雌、孕激素对二者表达的影响。方法:(1)用RT-PCR及SP免疫组化法检测51例异位内膜组织及28例正常子宫内膜组织中MMP-2/TIMP-2mRNA及蛋白的表达;(2)建立裸鼠子宫内膜异位症模型并予以雌、孕激素干扰,随机分成4组(E组、P组、E+P组及对照组C),观察不同激素干扰下裸鼠内异症模型的种植成功率及其大体、镜下的形态表现;用免疫组化法检测MMP-2/TIMP-2蛋白在不同组别裸鼠异位内膜中的表达。结果:(1)四组裸鼠内异症模型中种植物均成活,总种植成功率86.4%(51/59),各组间成功率无显著差异。其中P组种植物数量较其他组明显增多,P组、E+P组种植物明显大于E组、C组。受外源性雌、孕激素影响,镜下种植物分别呈现增生期和分泌期特点;(2)MMP-2/TIMP-2mRNA及蛋白在人及各组裸鼠异位内膜组织中均有表达,MMP-2相对表达量明显高于正常子宫内膜,差异有统计学意义(P<0.05);TIMP-2表达与正常内膜组织则无显著差异,MMP-2/TIMP-2比值明显增高(P均<0.01)。(3)E组、E+P组MMP-2蛋白表达均明显高于C组,而P组与C组无差异;TIMP-2蛋白在C组的表达明显高于其他激素干扰组。结论:裸鼠模型是EMs早期临床研究的理想模型。MMP-2/TIMP-2在人及裸鼠异位组织中异常表达,动态平衡紊乱,在EMs发生发展中起重要作用。雌激素可上调MMP-2蛋白表达,促进异位内膜种植;孕激素未表现对抗雌激素的作用,且通过抑制TIMP-2表达,同样造成MMP-2/TIMP-2比值升高,使异位内膜侵袭性增高,表现出"孕激素抵抗"现象。     

MMP-2 and TIMP-2 expression correlates with poor prognosis in cervical carcinoma--a clinicopathologic study using immunohistochemistry and mRNA in situ hybridization.

OBJECTIVE: The spread of malignant neoplasms is closely associated with matrix and basement membrane degradation, mediated by various classes of proteolytic enzymes. Matrix metalloproteinases (MMP) appear to have a key role in the sequence of events that lead to local invasion and metastasis. The present study evaluated the role of matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinases-2 (TIMP-2), and membrane-type metalloproteinase (MT1-MMP) in cervical neoplasia. METHODS: We have analyzed 49 uterine cervical squamous cell carcinomas, 10 cases of high-grade cervical intraepithelial neoplasia (CIN II-III), and 10 control cervices for the presence of MMP-2, TIMP-2, and MT1-MMP using in situ hybridization. MMP-2 protein expression was evaluated using immunohistochemistry. Results were analyzed for possible correlation with disease outcome. RESULTS: MMP-2, TIMP-2, and MT1-MMP mRNA were localized to both stromal and tumor cells. However, an intense signal for MMP-2 was detected almost exclusively in tumor cells and was uniformly absent from CIN lesions and control cervices. Conversely, intense signals for TIMP-2 and MT1-MMP were detected in both stromal and tumor cells of invasive carcinomas, more often for the former. As with MMP-2, they were absent from CIN lesions. MMP-2 protein expression was enhanced in tumor cells compared to CIN cases and controls, significantly compared to the latter (P = 0.01). The presence of both MMP-2 and TIMP-2 mRNA in tumor cells correlated with advanced stage (P = 0.003 for MMP-2, P = 0.002 for TIMP-2) and with poor survival (P = 0.003 for MMP-2, P = 0.002 for TIMP-2) in univariate analysis. In addition, their presence in tumor cells intercorrelated (P = 0.002). In multivariate survival analysis, MMP-2 presence retained its association with survival (P = 0.004), in addition to patient age (P = 0.027) and advanced stage (P = 0. 0002). CONCLUSIONS: Both MMP-2 and TIMP-2 have a key role in extracellular matrix invasion in cervical carcinoma, largely through their elaboration by tumor cells. The presence of mRNA for both proteins is interrelated and is associated with poor survival.