子宫内膜癌中基质金属蛋白酶及其组织抑制因子的表达与临床意义

目的研究基质金属蛋白酶(MMPs)- 2、9及其组织抑制因子(TIMPs)- 1、2在子宫内膜癌中的表达并探讨其与子宫内膜癌浸润转移的关系.方法应用免疫组织化学方法对121例子宫内膜癌组织及20例子宫脱垂妇女子宫内膜组织中MMP2、MMP9、TIMP1、TIMP2蛋白进行检测.结果MMP2、MMP9、TIMP1、TIMP2蛋白主要分布在内膜癌细胞中,在间质中也有少量表达.MMP2、MMP9、TIMP1、TIMP2在内膜癌细胞表达的阳性率均高于正常对照组.MMP2、MMP9蛋白的表达,在内膜癌细胞病理分级为G2、G3中,强阳性率高于G1者(P＜0.05);有肌层浸润内膜癌的强阳性率明显高于无肌层浸润者(P＜0.05);有淋巴转移者高于无淋巴转移者(P＜0.05);手术病理分期Ⅲ、Ⅳ期的强阳性率明显高于Ⅰ、Ⅱ期者(P＜0.05).TIMP1蛋白的表达在不同病理分级中差异无显著性(P＞0.05),在深肌层浸润内膜癌中阳性率低于浅肌层浸润(P＜0.05);有淋巴转移者的阳性率低于无淋巴转移者(P＜0.005),手术病理分期为Ⅲ、Ⅳ者阳性率低于Ⅰ期者(P＜0.05);TIMP2蛋白在不同病理分级、肌层浸润、淋巴转移和手术病理分期中的表达率比较,差异均无显著性(P＞0.05).结论MMP2、MMP9蛋白在内膜癌中的表达随病理分级而升高,随肌层浸润程度的加深及有淋巴转移而增加,TIMP1表达随肌层浸润程度的加深及有淋巴转移而下降.     

子宫内膜癌组织中FHIT、MMP-2的表达与临床病理的相关性研究

目的:探讨脆性组氨酸三联体(FHIT)和基质金属蛋白酶 2 (MMP- 2 )在子宫内膜癌组织中的表达及其与临床病理特征的关系。方法:免疫组化S P法检测12例正常子宫内膜,4 2例子宫内膜癌组织中FHIT、MMP- 2表达情况。结果:FHIT、MMP -2在正常子宫内膜和子宫内膜癌组织中的阳性表达率分别为10. 0 %、5 9.5 % ;4 1.7%、90 .5 % ,差异具有显著性(P =0 .0 11、P =0 .0 0 1)。在子宫内膜癌组织中,FHIT、MMP 2表达与子宫内膜癌的临床病理参数关系密切,表达强度与组织学病理分级、手术分期、肌层浸润程度和淋巴结转移相关(P <0 .0 1) ,FHIT、MMP -2表达存在负相关性(r=- 0 .6 5 7,P =0 .0 0 0 )。结论:联合检测FHIT、MMP 2在子宫内膜癌组织中的表达可为子宫内膜癌的早期诊断,进一步治疗及预后判断提供必要的理论依据。     

基质金属蛋白酶MMP-9及其组织抑制剂TIMP-1在卵巢肿瘤中的表达及意义

目的　研究基质金属蛋白酶MMP - 9及组织抑制剂TIMP - 1在卵巢肿瘤中的mRNA表达及与卵巢癌临床病理特征和预后的关系。方法　应用RT -PCR方法检测 4 8例卵巢癌、 2 1例良性卵巢肿瘤及 2 2例正常卵巢组织中MMP - 9及TIMP - 1mRNA的表达情况 ,进行阳性率及半定量的比较 ,并将其结果与临床及病理学资料进行统计学分析。结果　MMP - 9在卵巢癌中表达的阳性率及半定量明显高于正常卵巢组织 ,其表达水平与卵巢癌的手术病理分期及预后有关 ,在Ⅲ -Ⅳ期患者中的表达显著高于Ⅰ -Ⅱ期患者 (P <0 0 5 )。MMP - 9阳性患者累积生存率明显低于阴性者。TIMP - 1的表达在卵巢癌中无明显增高 ,且与临床病理无相关性。MMP - 9/TIMP- 1在卵巢癌中的比值明显高于正常卵巢组织。结论　MMP - 9及MMP - 9与TIMP - 1的平衡失调与卵巢癌的浸润转移密切相关 ,MMP - 9可作为监测晚期卵巢癌患者预后的一个指标。     

MMPs和TIMPs在卵巢上皮性肿瘤中表达的分析

目的　体内组织重建和生理过程中基质金属蛋白酶 (MMPs)和特异性抑制剂 (TIMPs)保持着动态平衡 ,不同的TIMP对不同的MMP具有不同的亲和性 ,本课题旨在研究MMP2、MMP9、TIMP1和TIMP2在卵巢上皮性肿瘤中的表达及其作用。方法　收集 1 996年 5月～ 2 0 0 1年 5月在浙江大学医学院附属妇产科医院行手术切除 ,并用福尔马林固定、石蜡包埋的 1 32例病例 ,运用免疫组织化学染色法检测。结果　MMP2、MMP9、TIMP1和TIMP2均表达于细胞胞浆内 ,均随着上皮性卵巢肿瘤恶性程度的增加 ,表达增强 ;在有淋巴结转移的患者中表达明显高于无淋巴结转移者 ;且MMP9 TIMP1、MMP2 TIMP2比例在恶性卵巢上皮性肿瘤中明显增高 ;相关性分析表明MMP2和TIMP2、MMP9和TIMP1间存在相关性 (P <0 0 0 0 1 )。结论　MMP2、MMP9、TIMP1和TIMP2与卵巢上皮性肿瘤的演化、侵袭过程关系密切     

基质金属蛋白酶MMP-9及其抑制剂TIMP-1在外阴癌中的表达

目的　观察基质金属蛋白酶MMP 9和其抑制剂TIMP 1在外阴癌组织中的表达情况 ,探讨其与外阴癌的浸润及转移的关系。方法　采用免疫组织化学SP方法对 33例外阴癌、2 3例VIN、15例外阴色素减退疾病、14例外阴尖锐湿疣和 12例正常皮肤组织进行标记及分析。结果　MMP 9、TIMP 1蛋白在以上 5组细胞浆中的阳性表达分别为 31/33、2 4 /33,2 1/2 3、19/2 3,15 /15、13/15 ,12 /14、12 /14 ,10 /12、7/12。MMP 9、TIMP 1蛋白在外阴癌、VIN、色素减退疾病、尖锐湿疣中的阳性表达率与正常对照组相比差异均无显著性意义 (P >0 0 5 ) ,但在(-～ +)组与 (++～ +++)组表达率之间MMP 9在外阴癌组与其他各组间相比差异均有显著性意义 (P<0 0 5 ) ;TIMP 1在外阴癌组与色素减退疾病组和正常组相比差异有显著性意义 (P <0 0 5 )。这 2个指标在外阴癌中的表达与临床病理特征无明显相关性。在外阴癌中MMP 9与TIMP 1在阳性表达之间差异有显著性意义 (P <0 0 5 ) ,而强阳性表达间差异有极显著性意义 (P <0 0 1)。结论　在外阴癌中MMP 9的表达强度超过TIMP 1时可能是外阴癌侵袭的一个早期指标 ,但不能单独作为预测外阴癌结局的一个标志。     

MMP-9及TIMP-1与胎膜早破关系的研究

目的 :研究胎膜早破 (PROM)孕妇羊水、血清中基质金属蛋白酶 9(MMP 9)、金属蛋白酶组织抑制剂 1(TIMP 1)的含量及在胎膜中的表达 ,探讨其与PROM的关系。方法 :选择临产前足月剖宫产分娩的胎膜早破者 2 5例 (PROM组 )和正常未破膜者 2 0例 (对照组 ) ,用双抗体夹心ELISA法测定羊水、血清中MMP 9、TIMP 1的含量 ,并用免疫组化SABC法检测胎膜中MMP 9及TIMP 1的表达。结果 :①PROM组和对照组羊水中MMP 9含量分别为 (887.2 4± 36 3.2 5 )与(6 6 6 .15± 2 39.5 8)ng/ml,差异有显著性 (P <0 .0 5 ) ;两组血清中MMP 9含量分别为 (38.72± 2 3.4 8)与 (17.4 5± 10 .77)ng/ml,差异有非常显著性 (P <0 .0 0 1) ;两组羊水与血清中TIMP 1含量差异无显著性 (P >0 .0 5 )。②PROM组血清与羊水中MMP 9、TIMP 1的含量无相关性 (r分别为 0 .133与 0 .10 3,P >0 .0 5 )。③PROM组和对照组胎膜MMP 9的阳性表达分别为 10 0 %、85 % ,差异有非常显著性 (P <0 .0 0 1)。结论 :胎膜早破孕妇羊水、血清中MMP 9含量明显升高 ,且MMP 9在胎膜中高表达 ,可能是胎膜早破发生的原因之一     

基质金属蛋白酶-9及其组织抑制剂TIMP-3在子宫内膜异位症的表达

目的 :探讨基质金属蛋白酶 9(MMP 9)及其抑制剂TIMP 3在子宫内膜异位症发生发展中的作用。方法 :采用免疫组化链霉菌抗生物素蛋白 过氧化物酶染色法 (SP法 )检测 4 3例子宫内膜异位症的异位内膜和在位内膜中MMP 9、TIMP 3的表达 ,以 19例正常子宫内膜为对照组 ,并用银染法观察腺上皮基底膜的完整性。结果 :在异位内膜组织的腺上皮细胞MMP 9呈高表达状态 ,与子宫内膜异位症在位内膜和正常子宫内膜相比 ,差异有高度显著性 (P <0 .0 1)。子宫内膜异位症的在位内膜和异位内膜TIMP 3均呈低表达 ,与正常内膜相比 ,差异有高度显著性 (P <0 .0 1) ,异位内膜较在位内膜TIMP 3的表达降低 ,差异有显著性 (P <0 .0 5 )。子宫内膜腺上皮的基底膜银染结果显示 :MMP 9与TIMP 3的染色强度比值与基底膜的阳性率呈负相关 (P <0 .0 1)。MMP 9、TIMP 3的表达强度与子宫内膜异位症的严重程度无显著相关性。结论 :异位内膜组织中MMP 9过度表达 ,而TIMP 3的表达下降 ,导致MMP 9/TIMP 3的比例增高 ,使子宫内膜异位症患者的异位内膜更具侵袭性 ,在子宫内膜异位症的发生发展中起重要作用     

子宫内膜癌PTEN蛋白表达及其临床意义

目的 :探讨抑癌基因第 10染色体同源丢失性磷酸酶 -张力蛋白基因 (PTEN)蛋白表达与子宫内膜癌的发生、发展及复发的关系。方法 :采用免疫组织化学S P法检测PTEN蛋白在 5 0例子宫内膜癌、12例不典型增生子宫内膜及 10例正常子宫内膜组织的表达。结果 :5 0例子宫内膜癌中PTEN蛋白表达率为 5 6 0 0 % ,明显低于正常子宫内膜以及不典型增生子宫内膜中的表达 (10 0 0 0 %、91 0 0 % ) (P <0 0 5 )。PTEN蛋白在无肌层浸润或肌层浸润≤ 1/ 2的子宫内膜癌中阳性表达率为 76 0 0 % ,肌层浸润 >1/ 2或有淋巴结转移者为 36 0 0 % ,与肌层浸润程度显著相关 (P <0 0 5 ) ,且Ia Ib期与Ic期相比也有统计学差异 ;但与子宫内膜癌的组织学分级、有无复发无关 (P >0 0 5 )。结论 :抑癌基因PTEN蛋白表达缺失在子宫内膜癌的发生、发展、浸润及复发过程中有一定作用。     

基质金属蛋白酶-2及孕激素受体在子宫内膜癌组织中的表达及其意义

目的检测基质金属蛋白酶-2（MMP-2）和孕激素受体（PR）在子宫内膜癌组织中的表达，分析其在子宫内膜癌中表达的相关性以及与临床病理特征的关系，探讨MMP-2在子宫内膜癌发生、发展中的作用。方法采用免疫组化SP法检测45例子宫内膜癌和12例增生子宫内膜中MMP-2、PR的表达，以20例正常子宫内膜作为对照。结果MMP-2蛋白在正常子宫内膜、增生子宫内膜中的阳性表达率比较，差异无显著性（P〉0．05）。子宫内膜癌组织中MMP-2阳性表达率（62．2％）高于对照组（15．6％）（P〈0．05）。MMP-2在中、低分化子宫内膜癌组织中的阳性表达率（82．6％）高于高分化子宫内膜癌组织（40．9％）（P〈0．01），有肌层浸润标本中的阳性表达率（71．1％）高于无肌层浸润者（14．3％）（P〈0．05）；绝经者（65．5％）与未绝经者（56．3％）、手术病理为Ⅰ期者（52．0％）与Ⅱ～Ⅳ者（75．0％）以及有淋巴结转移者（63．6％）与无淋巴结转移者（61．8％）相比，MMP-2的阳性表达率差异无显著性。子宫内膜癌中PR的阳性表达率（55．6％）明显低于对照组（84．4％）（P〈0．05）。MMP-2与PR在子宫内膜癌中的表达呈负相关（P=0．0048，r=-0．42）。结论MMP-2在子宫内膜癌中呈高表达，可能在其进展中有重要作用，PR表达降低可能是MMP-2表达升高的原因；检测MMP-2在子宫内膜癌中的表达，有助于子宫内膜癌预后的判断。     

子宫内膜癌中基质金属蛋白酶-9表达及其临床意义

目的研究基质金属蛋白酶(MMPs)-9在子宫内膜癌中的表达以及临床意义。方法对广州医学院第三附属医院1992-1997年收治的128例子宫内膜癌、30例不典型增生内膜及30例正常子宫内膜石蜡标本,应用免疫组织化学方法检测MMP-9蛋白表达情况。结果子宫内膜癌组织与子宫内膜不典型增生、正常子宫内膜组织相比,MMP-9蛋白的阳性表达率增加,差异有统计学意义(χ2=8.154,P<0.05)。MMP-9蛋白阳性表达与子宫内膜癌病理分级及淋巴转移有关(P<0.05)。结论 MMP-9蛋白在子宫内膜癌的浸润转移中起促进作用,可能与子宫内膜癌的预后不良有关。     

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.     

Anti-invasive effect of MMI-166, a new selective matrix metalloproteinase inhibitor, in cervical carcinoma cell lines

OBJECTIVES: The aim of our study was to evaluate the anti-invasive effect of MMI-166, a new matrix metalloproteinase (MMP) inhibitor in cervical carcinoma cell lines. METHODS: We analyzed the invasive activities of cervical carcinoma cell lines (CAC-1, CaSki, and SiHa) and the gene expression of various matrix proteinases (matrix metalloproteinase-1 [MMP-1], MMP-2, MMP-9, membrane-type MMP type 1 [MT1-MMP], MT2-MMP, and MT3-MMP) and their inhibitors (tissue inhibitor of metalloproteinase type 1 [TIMP-1] and TIMP-2). The effect of MMI-166 was analyzed by in vitro invasion assay. The cytotoxicity of MMI-166 was determined by MTT assay. The gelatinase activity was analyzed by gelatin zymography. RESULTS: Cervical carcinoma cell lines, which produced both MMP-2 and MT1-MMP, showed invasive capacity in the in vitro invasion assay. The invasion of cervical carcinoma cells was suppressed by MMI-166. No remarkable suppression of the proliferation by MMI-166 was observed in the MTT assay. Gelatin zymography revealed complete suppression of MMP-2 activity by MMI-166. CONCLUSIONS: MMI-166 inhibited the MMP-2 activity in cervical carcinoma cells and it is useful for the regulation of cervical carcinoma cell invasion.     

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。     

Inhibitory effect of ginsenoside-Rb2 on invasiveness of uterine endometrial cancer cells to the basement membrane.

Ginsenoside-Rb2 derived from ginseng inhibited invasiveness to the basement membrane of endometrial cancer cell lines Ishikawa. HHUA and HEC-1-A cells. These cells dominantly expressed matrix metalloproteinase (MMP)-2 (gelatinase A) among MMPs by zymography. Ginsenoside-Rb2 suppressed the expression and activity of MMP-2, but did not alter the expression of tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 in the cells. Therefore, ginsenoside-Rb2 might inhibit invasiveness to the basement membrane via MMP-2 suppression in some endometrial cancers, and can be used as a medicine for inhibition of secondary spreading of uterine endometrial cancers.     

Expression of matrix metalloproteinase-26 and tissue inhibitors of metalloproteinases TIMP-3 and -4 in benign endometrium and endometrial cancer

OBJECTIVE: Matrix metalloproteinases (MMPs) and their physiological inhibitors, the tissue inhibitors of MMPs (TIMPs), play a key role in tumor cell invasion, angiogenesis, and growth. The aim of this study was to determine the expression and cellular distribution of MMP-26, TIMP-3, and TIMP-4 in endometrial cancers and benign endometrium throughout the menstrual cycle and the correlation with tumor histological subtype, stage, and grade. METHODS: Immunohistochemical analysis using polyclonal antibodies generated against pro- and active MMP-26, and mono- and polyclonal antibodies specific to TIMP-3 and TIMP-4, respectively, was performed. RESULTS: MMP-26, TIMP-3, and TIMP-4 are expressed in endometrial carcinomas (N = 86) and benign endometrium (N = 50) from various stages of the menstrual cycle. Semi-quantitative analysis of staining intensity indicated that endometrial carcinomas expressed more MMP-26, TIMP-3, and TIMP-4 compared to benign endometrium from the postmenopausal period, but not from the secretory phase of the menstrual cycle. The highest staining intensity was associated with endometrial epithelial cells, followed by vascular endothelial cells, myometrial smooth muscle cells, and endometrial stromal cells. Increased staining intensity of MMP-26 and TIMP-3 correlated with grade III tumors and MMP-26 and TIMP-4 with the depth of myometrial invasion in tumors histologically characterized as endometrioid adenocarcinoma, clear-cell, and papillary serous carcinoma staged/graded based on FIGO criteria. CONCLUSION: MMP-26 and TIMP-4 are expressed in endometrium and endometrial carcinoma and their elevated expression and correlation with myometrial invasion suggests that MMP-26 and TIMP-4 may play a key role in endometrial tumor progression.     

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.     

Expression and localization of matrix metalloproteinases (MT1-MMP, MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) during synepitheliochorial placentation of goats (Capra hircus)

Matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) play key roles during the placentation of highly invasive haemochorial type. Our knowledge is yet scanty, however, regarding the roles played by MMPs and TIMPs in the placentation of non-invasive synepitheliochorial type. In the present study, expression patterns of MT1-MMP, MMP-2 and TIMP-2 mRNAs as well as the encoded proteins in the endometrium and the placenta were examined on Days 35, 75, and 100 of pregnancy, representing roughly the 1st, 2nd and 3rd trimesters of caprine gestation, by means of quantitative RT-PCR analysis, in situ hybridization, immunoblotting, gelatin zymography and immunohistochemistry. In the endometrium and the intercotyledonal trophoblast, the expression levels of the 3 genes remained relatively uniform throughout the period of gestation examined. Curiously, however, in the placentomes, the relative expression levels of MT1-MMP mRNA increased linearly from Day 35 to Day 100, while those of MMP-2 and TIMP-2 were clearly down-regulated in Day 100 placentae. The expression levels of MT1-MMP and TIMP-2 proteins in placentomes were well correlated with those of the respective mRNAs. In the case of MMP-2, the total amount of MMP-2 protein (the combined values of the latent, the intermediate and the active forms) decreased slightly, while the levels of the active form increased markedly from Day 35 to Day 100. Immunohistochemical analysis of the placentome revealed that MT1-MMP and TIMP-2 proteins were co-localized in the binucleate trophoblast cells; expression of these 2 proteins was not detected in the uninuclear principal trophoblast cells. MMP-2 expression was detected both in the binucleate and in the uninuclear principal cells of the trophoblast and in the endometrial stromal cells of the uterine septum, regardless of the stages of gestation examined. The co-localization of MT1-MMP, MMP-2 and TIMP-2 in binucleate trophoblast cells, the cotyledonal trophoblast cells and the subsyncytial stromal cells is likely to reflect the functional coordination of the 3 proteins in these cells during trophoblastic invasion and the placental tissue remodeling in the placentome.     

Expression of matrix metalloproteinases and related tissue inhibitors in the cyst fluids of ovarian mucinous neoplasms

OBJECTIVES AND METHODS: The growth of an ovarian cystic neoplasm often involves its invasion into and destruction of the extracellular matrix. We examined neoplastic cysts of ovarian mucinous tumors for the presence of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) using zymography (in situ zymography, gelatin zymography, and casein zymography) and enzyme-linked immunosorbent assay. RESULTS: Matriolytic activity was detected within the cystic contents and cytoplasm of the lining epithelial cells of the cyst by in situ zymography. This intracystic matriolytic activity was thought to originate mainly in the epithelial cells. The activated form of MMP-9 was seen in all carcinoma and borderline fluids and in 7 of 15 adenomas. The concentration of MMP-9 was higher in carcinoma fluids than in borderline and adenoma fluids (P < 0.05). TIMP-1, which specifically binds to MMP-9, was also higher in carcinoma and borderline fluids than in adenoma fluids (P < 0.05). MMP-2 activity was nearly ubiquitously present in all cyst fluids, irrespective of the fluid's histologic category. The amount of MMP-2 was highest in the carcinoma category, although not to a statistically significant degree. TIMP-2, a specific inhibitor for MMP-2, was significantly lower in the borderline category than in the adenoma category. The molar ratios of TIMP-1/MMP-9 (not significant) and TIMP-2/MMP-2 (P < 0.05) were higher in the adenoma category. Expressions of trypsin, MMP-7, and MMP-9 were generally higher in carcinoma and borderline fluids than in adenoma fluids. CONCLUSIONS: These observations indicate the importance of ovarian cystic fluids for analyzing tumor-associated matriolytic activities. The findings suggest that these matriolytic enzymes, together with the presence of their inhibitors, play an important role in the growth of ovarian mucinous tumors.     

RECK与MMP-9基因在子宫内膜癌中的表达及临床意义

目的:研究RECK和MMP-9在不同子宫内膜组织中的表达,探讨两者在子宫内膜癌的发生、发展和浸润转移中的作用。方法:应用链霉菌抗生物素蛋白-过氧化酶(SP)免疫组化法检测42例子宫内膜癌组织、15例子宫内膜不典型增生组织及26例正常增生期子宫内膜组织中RECK蛋白及MMP-9蛋白表达情况。结果:与正常增生期子宫内膜组织及子宫内膜不典型增生组织相比,子宫内膜癌组织中RECK蛋白阳性表达率显著降低(χ2=9.307,P<0.05),MMP-9蛋白阳性表达率显著增高(χ2=11.438,P<0.05),RECK蛋白与MMP-9蛋白在子宫内膜癌中的表达存在明显负相关(P<0.01)。RECK蛋白的表达水平与临床分期、组织学分级、肌层浸润深度及淋巴结转移密切相关(均P<0.05);MMP-9蛋白的表达水平与临床分期、组织学分级、肌层浸润深度密切相关(均P<0.05),而与淋巴结转移无关。结论:RECK蛋白的表达缺失和MMP-9的表达可能与子宫内膜癌的发生、发展及浸润转移有关。     

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.