检测基质金属蛋白酶及其抑制物对判断膀胱癌侵袭和转移的意义

目的 :研究基质金属蛋白酶 (MMPs)中MMP 2、MMP 9及MMP 9抑制物TIMP 1在膀胱移行细胞癌中的表达情况及其在判断膀胱癌侵袭转移中的作用。方法 :应用免疫组化LSAB法检测 90例膀胱移行细胞癌 (transitionalcellcarcinomaofbladder,TCCB)组织中MMP 2、MMP 9和TIMP 1蛋白表达水平。结果 :膀胱癌组织中MMP 2、MMP 9和TIMP 1蛋白表达显著高于正常膀胱黏膜。浸润性膀胱癌组织中阳性表达显著高于表浅性膀胱癌组织。阳性表达细胞在肿瘤 -间质界面较多。与病理组织学分级、分期均呈正相关 ;与肿瘤的复发和转移呈正相关。结论 :MMP 2、MMP 9和TIMP 1参与膀胱癌侵袭转移 ,在膀胱移行细胞癌中的表达具有预后价值。     

双调蛋白诱导乳腺癌细胞表达尿激酶型纤溶酶原活化物

目的:在人乳腺癌模型上研究双调蛋白与尿激酶型纤溶酶原活化物表达之间的关系.方法:乳腺癌NS2T2A1细胞经双调蛋白反义cDNA质粒转染后经潮霉素B筛选获得表达双调蛋白反义RNA的AR-AS1及AR-AS3两个细胞克隆,转染空载体获得NS2T2A1 V对照细胞,接种至裸鼠皮下形成肿瘤.测定细胞及肿瘤uPA表达水平,并研究uPA与细胞侵袭性之间的关系.结果:AR-AS1及AR-AS3细胞体外及体内uPA表达均被抑制.外源性双调蛋白可刺激对照细胞uPA的表达,并部分恢复AR-AS1及AR-AS3细胞uPA的表达水平.双调蛋白反义cDNA质粒转染及抗uPA抗体均导致乳腺癌细胞体外侵袭性的降低.结论:在乳腺癌模型上,uPA表达与肿瘤细胞的侵袭密切相关.双调蛋白反义RNA表达可有效地抑制uPA的表达,进而抑制肿瘤细胞的侵袭性.     

检测基质金属蛋白酶及其抑制物对判断膀胱癌侵袭和转移的意义

目的：研究基质金属蛋白酶(MMPs)中MMP—2、MMP—9及MMP—9抑制物TIMP—1在膀胱移行细胞癌中的表达情况及其在判断膀胱癌侵袭转移中的作用。方法：应用免疫组化LSAB法检测90例膀胱移行细胞癌(transitional cell carcinoma of bladder,TCCB)组织中MMP—2、MMP—9和TIMP—1蛋白表达水平。结果：膀胱癌组织中MMP—2、MMP—9和TIMP—1蛋白表达显著高于正常膀胱黏膜。浸润性膀胱癌组织中阳性表达显著高于表浅性膀胱癌组织。阳性表达细胞在肿瘤—间质界面较多。与病理组织学分级、分期均呈正相关；与肿瘤的复发和转移呈正相关。结论：MMP—2、MMP—9和TIMP—1参与膀胱癌侵袭转移，在膀胱移行细胞癌中的表达具有预后价值。     

UTI抑制骨肉瘤细胞侵袭与转移的实验研究

目的体外研究尿胰蛋白酶抑制剂(urinary trypsininhibitor,UTI)抑制人骨肉瘤细胞系MG-63细胞的侵袭与转移。方法在体外培养的MG-63细胞培养基中加入不同浓度的UTI,对照组中加入50μl PBS缓冲液,用免疫组化、RT-PCR和Western blot方法检测尿激酶型纤溶酶原激活物(urokinasetype plasminogen activator,UPA)的表达情况。结果免疫组化染色、RT-PCR及Western blot检测发现随着UTI浓度的逐渐增加,UPA表达量逐渐减少,当UTI浓度增加到100nM时,抑制作用明显(P<0.05),并且呈剂量依赖性。结论UTI不但从mRNA水平,而且从蛋白质水平抑制骨肉瘤细胞表达UPA,从而抑制骨肉瘤细胞侵袭与转移。     

miR-623在膀胱癌中的表达及靶向Fascin1抑制膀胱癌细胞迁移和侵袭

目的:探讨miR-623在膀胱癌中的表达及通过靶向Fascin1对膀胱癌细胞迁移和侵袭能力的影响。方法:采用实时荧光定量PCR检测正常膀胱上皮组织、膀胱癌组织、正常永生化膀胱上皮细胞系(SV-HUC-1)和膀胱癌细胞系（T24、UMUC3）中miR-623的表达量;采用脂质体瞬时转染miR-623 mimics,划痕实验和Transwell侵袭实验检测miR-623过表达后膀胱癌细胞迁移、侵袭能力的改变;生物信息学预测miR-623的作用靶蛋白。miR-623过表达后Western blot及双荧光素酶报告基因检测其靶点的表达及结合情况;使用Fascin1特异性siRNA观察膀胱癌细胞迁移和侵袭能力的变化,并同时转染miR-623 inhibitor进行恢复实验。结果:miR-623在膀胱癌中的表达水平显著低于在正常膀胱组织中的表达（P＜0.05）,在膀胱癌细胞系（T24、UMUC3）中的表达水平显著低于正常膀胱细胞系(SV-HUC-1)（P＜0.05）;miR-623过表达显著抑制T24和UMUC3细胞的迁移和侵袭能力。生物信息学预测Fascin1为miR-623的靶基因,在T24和UMUC3细胞中过表达miR-623,能够显著降低Fascin1的蛋白水平;荧光素酶报告基因分析结果证实miR-623作用于Fascin1的3'-UTR。下调Fascin1表达能够抑制膀胱癌T24和UMUC3细胞的迁移和侵袭能力,同时抑制miR-623的表达能够提高细胞的迁移和侵袭能力。结论:miR-623在膀胱癌中表达水平降低,是一个抑癌因子;并可能通过靶向Fascin1调节膀胱癌的侵袭和转移能力。     

CD24在复发性膀胱癌中的表达及其对癌细胞增殖的影响

目的：分析 CD24在复发性膀胱癌组织中的表达,探讨抑制 CD24表达对膀胱癌细胞增殖的影响。方法：应用免疫组化检测36例复发性膀胱癌组织中 CD24的表达情况,并进行半定量分析。利用脂质体转染法将含有 CD24特异性 siRNA 的真核表达质粒转染至人膀胱癌 BIU －87细胞中,实时定量 PCR 和 Western blot 法检测 BIU －87细胞中 CD24的表达情况改变,MTT 法、Transwells 侵袭实验分别检测下调 CD24的表达后,膀胱癌细胞增殖、侵袭能力发生的变化。并分析细胞中信号转导及转录活化因子（STAT3）、细胞周期蛋白（CyclinD1）、基质金属蛋白酶（MMP －9）的表达变化。结果：CD24在复发性膀胱癌组织中高表达,高级别膀胱癌组织中,阳性率达76．2％。细胞学实验中,经实时定量 PCR 和 Western blot 法证实实验组较对照组细胞CD24 mRNA 转录和蛋白表达水平明显降低。MTT 检测发现实验组较对照组细胞的增殖能力显著下降;Tran-swells 侵袭实验显示实验组的穿膜细胞数显著少于对照组。实验组中 STAT3、CyclinD1、MMP －9表达有明显下降。结论：CD24在复发性膀胱癌中高表达,并与肿瘤的分级呈正相关。体外实验中,抑制 CD24基因表达能显著抑制膀胱癌细胞的增殖和侵袭能力,间接提示 CD24可能参与膀胱癌的复发进程。     

uPA、uPAR与尿路移行细胞癌侵袭行为的相关性研究

背景与目的:丝氨酸蛋白水解酶的uPA(urokinase-typeplasminogenactivator)uPAR(urokinase-typeplasminogenactivatorreceptor)作用系统被认为在细胞外基质和基底膜的降解、促进肿瘤侵袭转移过程中起核心作用,本研究旨在探讨uPA、uPAR在尿路移行细胞癌组织中的表达及其与肿瘤侵袭行为的关系。方法:应用免疫组织化学PicTureTM通用型二步法检测50例肾盂输尿管癌及40例膀胱癌组织中uPA和uPAR的表达水平及分布。结果:uPA和uPAR在正常肾盂、输尿管、膀胱组织中未见表达,在癌组织中的表达明显高于癌旁组织;G1级uPA和uPAR的阳性率分别为33.33%、50.00%,G3级分别为88.47%和96.15%;Ta～T1期的分别为37.50%和50.00%,T4期均为100.0%;在高分级、高分期肿瘤组织中,uPA和uPAR的阳性率明显升高(P<0.05),且uPA阳性与uPAR阳性高度相关(rs=0.979)。结论:uPA、uPAR的共同表达是尿路移行细胞癌的特征之一,且与分级和分期密切相关,在尿路移行细胞癌浸润和转移中可能具有重要作用。     

CBX7基因与膀胱癌转移的相关性及其对细胞侵袭能力影响

目的:探讨CBX7基因与膀胱癌转移的相关性及其过表达对膀胱癌细胞侵袭能力的影响。方法:应用荧光定量PCR检测膀胱癌组织中CBX7 mRNA的表达。构建CBX7真核表达载体pc DNA-CBX7,转染膀胱癌T24细胞,检测转染后CBX7蛋白的表达以及T24细胞的侵袭能力。结果:CBX7基因在膀胱癌组织中明显下调,而且侵袭性膀胱癌组织中CBX7的表达显著低于浅表性膀胱癌;pc DNA-CBX7转染后能够显著升高T24细胞CBX7的表达量;CBX7过表达的T24细胞穿透滤膜的数量明显减少。结论:CBX7基因与膀胱癌的转移相关,其过表达能够降低T24细胞的侵袭能力。     

尿激酶氨基末端基因抗乳腺癌细胞转移的实验研究

目的：探讨尿激酶氨基末端（ATF）基因转移对肿瘤转移的抑制作用。方法：构建重组ATF基因真核表达载体pcDNA3-ATF,用脂质体Lipofectin介导,将其导入其证明尿激酶（uPA)和尿激酶受体（uPAR）均有表达的人乳腺癌细胞系MCF－7,用Western blot检测转染细胞ATF基因的表达,体外观察并比较了野生型和转基因MCF－7细胞侵袭人工基底膜能力;裸鼠皮下接种癌细胞,复制自发性肿瘤转移模型,观察成瘤性和转移性。结果uPA／uPAR在MCF－7中有较高水平的表达;转ATF基因后,可检测到ATF的明显表达,体外侵袭穿透人工基底膜的能力明显降低,体内原位成瘤性不受影响,但自主性体肺转移能力有一定程度的下降,结论ATF基因转移后,ATF的表达可能竞争 抑制内源性uPA与uPAR的结合,在一定程度上抑制癌细胞的侵袭和转移。     

人膀胱癌组织中环氧化酶2的表达

背景与目的：环氧化酶（cyclooxygenase,COX)是人体内合成前列腺素的限速酶。最近研究表明,环氧化酶2（COX－2）与肿瘤的生成有关。本研究通过检测COX－1和COX－2在人膀胱癌组织、正常膀胱粘膜以及膀胱炎症组织中的表达,探讨COX在膀胱癌发生发展中的作用。方法：应用逆转录聚合酶链反应（RT－PCR）和免疫组化法（Envision二步法）,检测膀胱移行细胞癌和癌旁组织、正常膀胱粘膜以及膀胱炎症组织中COX－1和COX－2 mRNA和蛋白的表达,并分析癌组织中COX的表达强度与肿瘤对应的各项病理参数之间的关系。结果：RT－PCR检测15例新鲜膀胱癌组织COX－2 mRNA均阳性表达,5例肉眼所见的癌旁正常组织中仅1例阳性,两者差异有显著性;而COX－1 mRNA在所有新鲜癌组织标本中均有结构性表达。免疫组织化学研究结果与RT－PCR结果相似,COX－2蛋白主要集中在肿瘤细胞浆内,阳性表达率为50％,正常膀胱粘膜（n=4)和膀胱慢性炎症组织（n=5)中没有表达;反之,COX－1蛋白主要表达在正常或炎症组织的平滑肌细胞上,肿瘤组织中为阴性表达。在40例膀胱移行细胞癌石蜡切片标本中,COX－2蛋白的表达强度与肿瘤的分级和分期有关,恶性度较高的Ⅲ级癌的表达水平高于I级和Ⅱ级癌,浸润性癌（T2－4）也高于浅表性癌（Ta-1)。结论：COX－2 mRNA和蛋白在人膀胱癌组织中表达增高,并与肿瘤的恶性率相关,说明COX－2可能在膀胱癌的发生发展中起着重要作用。     

Targeting of urokinase plasminogen activator receptor in human pancreatic carcinoma cells inhibits c-Met- and insulin-like growth factor-I receptor-mediated migration and invasion and orthotopic tumor growth in mice

Pancreatic carcinomas express high levels of urokinase-type plasminogen activator (uPA) and its receptor (uPAR), both of which mediate cell migration and invasion. We investigated the hypotheses that (a) insulin-like growth factor-I (IGF-I)- and hepatocyte growth factor (HGF)-mediated migration and invasion of human pancreatic carcinoma cells require uPA and uPAR function and (b) inhibition of uPAR inhibits tumor growth, retroperitoneal invasion, and hepatic metastasis of human pancreatic carcinomas in mice. Using transwell assays, we investigated the effect of IGF-I and HGF on L3.6pl migration and invasion. We measured the induction of uPA and uPAR following treatment of cells with IGF-I and HGF using immunoprecipitation and Western blot analysis. The importance of uPA and uPAR on L3.6pl cell migration and invasion was studied by inhibiting their activities with amiloride and antibodies before cytokine treatment. In an orthotopic mouse model of human pancreatic carcinoma, we evaluated the effect of anti-uPAR monoclonal antibodies with and without gemcitabine on primary tumor growth, retroperitoneal invasion, and hepatic metastasis. IGF-I and HGF mediated cell migration and invasion in L3.6pl cells. In addition, IGF-I and HGF induced uPA and uPAR expression in L3.6pl cells. In vitro, blockade of uPA and uPAR activity inhibited IGF-I- and HGF-mediated cell migration and invasion. Treatment of mice with anti-uPAR monoclonal antibody significantly decreased pancreatic tumor growth and hepatic metastasis and completely inhibited retroperitoneal invasion. Our study shows the importance of the uPA/uPAR system in pancreatic carcinoma cell migration and invasion. These findings suggest that uPAR is a potential target for therapy in patients with pancreatic cancer.     

Short interfering RNA directed against Slug blocks tumor growth, metastasis formation, and vascular leakage in bladder cancer

In a previous study, we have identified Slug as selectively overexpressed in the highly invasive bladder cancer cells. Furthermore, Slug overexpression was correlated with metastasis in bladder cancer tissues. Here, we investigated the role and underlying mechanism of Slug in the process of growth, invasion and metastasis formation, and vascular leakage in a bladder cancer cell line T24. We knocked down in a bladder cancer cell line T24 by Slug siRNA, and the effects on invasion, metastasis, and proliferation were evaluated in vitro. A pseudometastatic model of bladder cancer in severe combined immunodeficient mice was used to assess the effects of Slug silencing on metastasis and angiogenesis development. Slug-specific siRNA in T24 cells led to a significant decrease in invasiveness and metastasis, proliferation rates, and angiogenesis in vitro. Slug-specific siRNA induced strong changes in cell invasiveness mainly through a mechanism of up-regulation of epithelial markers E-cadherin expression. Interestingly, E-cadherin-specific siRNA attenuated Slug siRNA-induced Matrigel invasion. Moreover, Slug-specific siRNA induced strong changes in microvessel counts in angiogenesis mainly through decreased activity of MMP-2, but not through cadherin expression and decreased activity of MMP-9. Restoration of MMP-2 expression in the Slug-silenced T24 cells resulted in an increased cell angiogenesis. Moreover, Slug-specific siRNA significantly reduced tumor growth by approximately 60% and inhibited metastasis and angiogenesis in a xenograft mouse model. This was accompanied by an increased expression of E-cadherin and a decreased expression of MMP-2 in tumor tissues. These data suggest that Slug regulates tumor growth, invasion and metastasis, and angiogenesis through E-cadherrin and MMPS passway. Blocking Slug signaling in bladder cancer may represent a novel strategy in the future to reduce metastatic disease burden in bladder cancer patients.     

miRNA-20a在膀胱癌中的表达及其机制研究

  目的  探讨miRNA-20a在膀胱癌组织中的表达及其机制。  方法  收集2014年1月至2015年1月昆明医科大学第二附属医院96例患者的组织标本,运用实时定量聚合酶链反应（qRT-PCR）方法检测miRNA-20a在膀胱癌组织和癌旁组织中的表达;生物信息学方法预测miRNA-20a的靶基因并采用双荧光素酶报告基因实验进行验证;qRT-PCR、Western blot以及细胞免疫荧光分别检测人膀胱癌细胞系T24和J82细胞转染miRNA-20a模拟物或阴性对照后对靶基因mRNA和蛋白表达的影响;CCK-8、Transwell侵袭小室和划痕实验检测过表达miRNA-20a后T24细胞体外增殖、迁移和侵袭能力的改变。  结果  miRNA-20a在膀胱癌组织中高表达,且与肿瘤的病理分级、临床分期、转移以及复发密切相关（P＜0.001）;双荧光素酶报告基因实验证实miRNA-20a与人源性长寿保障基因2（Homo sapiens longevity assurance homologue 2,LASS2）的3'-UTR直接靶向结合;转染miRNA-20a模拟物可显著下调膀胱癌细胞中LASS2 mRNA和蛋白的表达（P＜0.01）,增强膀胱癌细胞的增殖、侵袭和迁移能力（P＜0.01）。  结论  miRNA-20a在膀胱癌组织中高表达,且miRNA-20a可通过靶向抑制LASS2促进膀胱癌细胞的增殖、侵袭和迁移,与膀胱癌的发生发展相关。      

Expression of urokinase and urinary trypsin inhibitor in metastatic and non-metastatic murine Lewis lung carcinoma cells

The murine Lewis lung carcinoma 3LL cells give rise to spontaneous and experimental lung metastasis in C57BL/6 mice. Tumor cells maintained by serial subcutaneous transplantation in mice retain their ability to form lung metastasis, while cells carried in vitro loose metastatic potential with time. In order to obtain the non-metastatic subline, 3LL cells selected for its high lung colonization potential was grown continuously in vitro for 24 weeks. The present study was undertaken to characterize the expression of both urokinase-type plasminogen activator (uPA) and urinary trypsin inhibitor (UTI) in the non-metastatic (3LL(-)) and the metastatic (3LL(+)) cells. Both cells were tested on the Matrigel for invasive ability using a modified Boyden chamber and assayed for expression of uPA and UTI. The 3LL(+) cells secreted 5 times more uPA (6.25 mu g per 10(6) cells per 24 h) than the 3LL(-) cells (1.25 mu g per 10(6) cells per 24 h). The 3LL(+) cells, which expressed 2 times more cell-surface receptor-bound enzymatically active uPA (0.32 +/- 0.06 OD405) than the 3LL(-) cells (0.15 +/- 0.03 OD405), had larger amounts of cell-surface receptor-bound uPA. On the other hands, UTI levels in the conditioned media was decreased 25-fold in the 3LL(+) cells (0.05 mu g/10(6) cells/24 h) compared to the 3LL(-) cells (1.25 mu g/10(6) cells/24 h). The 3LL(-) cells expressed significantly higher levels of cell-associated UTI as indicated by a cell ELISA (3LL(+), 0.30 +/- 0.04 OD450; 3LL(-), 1.30 +/- 0.21 OD450) and by Western blot analysis. Metastatic competence in the 3LL(+) tumor model is associated with increased expression and release of uPA, as well as decreased UTI production, consistent with a more invasive phenotype. These data support our hypothesis that UTI may contribute to the inhibition of uPA expression in tumor cells.     

Demethylation-linked activation of urokinase plasminogen activator is involved in progression of prostate cancer

Increased expression of urokinase plasminogen activator (uPA) has been reported in various malignancies including prostate cancer. However, the mechanism by which uPA is abnormally expressed in prostate cancer remains elusive. Here, we show that uPA is aberrantly expressed in a high percentage of human prostate cancer tissues but rarely expressed either in tumor-matched nonneoplastic adjacent tissues or benign prostatic hyperplasia samples. This aberrant expression is associated with cancer-linked demethylation of the uPA promoter. Furthermore, treatment with demethylation inhibitor S-adenosylmethionine or stable expression of uPA short hairpin RNA significantly inhibits uPA expression and tumor cell invasion in vitro and tumor growth and incidence of lung metastasis in vivo. Collectively, these findings strongly suggest that DNA demethylation is a common mechanism underlying the abnormal expression of uPA and is a critical contributing factor to the malignant progression of human prostate tumors.     

LIM kinase 1 increases tumor metastasis of human breast cancer cells via regulation of the urokinase-type plasminogen activator system

Mammalian LIM kinase 1 (LIMK1) phosphorylates and inactivates the actin-binding and -depolymerizing factor cofilin and induces actin cytoskeletal changes. LIMK1 is reported to play an important role in cell motility, but the mechanism of induction of cell motility and the role of LIMK1 in tumor growth, angiogenesis and invasion are poorly understood. Here we show that expression of LIMK1 in MDA-MB-435 human breast cancer cells enhanced cell proliferation and cell invasiveness and promoted in vitro angiogenesis. Since tumor metastasis requires degradation of the extracellular matrix by the serine protease urokinase type plasminogen activator (uPA), we examined the role of LIMK1 in the regulation of uPA/uPAR system. LIMK1 overexpression in breast cancer cells upregulated the uPA system, increased uPA promoter activity, induced uPA and uPAR mRNA and protein expression and induced uPA secretion. In contrast, cells transfected with the catalytically inactive LIMK mutant D460N-LIMK1 did not exhibit these phenotypic changes. Blocking antibodies against uPA and uPAR suppressed LIMK1-induced cell invasiveness. In addition, LIMK1 overexpression increased tumor growth in female athymic nude mice, promoted tumor angiogenesis and induced metastasis to livers and lungs, possibly by increasing uPA expression in the tumors. Finally, LIMK1 and uPAR were coordinately overexpressed in human breast tumors. These results suggested an important role for LIMK1 signaling in breast cancer tumor growth, angiogenesis and invasion and a regulatory connection between LIMK1 and the uPA system.     

Pleiotrophic inhibition of pericellular urokinase-type plasminogen activator system by endogenous tumor suppressive maspin.

Maspin is a novel serine protease inhibitor with tumor suppressive activity, inhibiting tumor invasion and metastasis. To date, the underlying molecular mechanism of maspin remains elusive. Recombinant maspin has been shown to specifically inhibit cell surface-associated urokinase-type plasminogen activator (uPA) and fibrinogen-bound tissue-type plasminogen activator. However, the role of endogenous maspin in plasminogen activation is totally unknown. To address this issue, we generated stable maspin-expressing transfectants using prostate carcinoma cells DU145 as the parental cell line. We report here that endogenous maspin exerts pleiotropic inhibitory effects on the pericellular uPA system. Maspin expression led to a significantly reduced level of cell surface-bound uPA and uPA receptor proteins without altering the steady-state levels of the respective mRNAs. Treatment with receptor-associated protein (RAP), a specific inhibitor of low-density lipoprotein receptor-related protein, lead to a significantly increased level of secreted uPA and cell surface uPAR in maspin transfectants but not in the mock control cells. A combination of enzymatic and molecular analyses revealed that maspin inhibits the cell surface-mediated plasminogen activation by forming an SDS-resistant complex with cell surface-bound uPA. In addition, maspin expression led to a dramatic reduction in the release of active uPA, both high molecular weight and the low molecular weight, into the conditioned culture medium. Consistently, the conditioned medium of maspin transfectant clones had a significantly reduced activity in converting plasminogen to plasmin. The inhibitory effect of maspin on pericellular uPA correlates with significantly decreased cell invasion potential and motility in vitro. The maspin-neutralizing antibody (Abs4A) reversed the subdued invasive potential of maspin transfectant cells in a dose-dependent manner. In summary, this study provides the first evidence that endogenous maspin is a potent inhibitor of pericellular uPA. Furthermore, our results support a current hypothesis that maspin blocks tumor invasion and motility by inhibiting localized pericellular proteolysis.     

Doxycycline抑制大肠癌LS174T细胞侵袭的体外实验研究

目的:观察多西环素(Doxycycline)在体外对LS174T侵袭能力的抑制作用,并尝试探讨该抑制作用的分子机制。方法:1)MTT法观察Doxycycline对LS174T细胞粘附力的影响;Transwell小室法检测不同浓度Doxycycline对细胞侵袭和运动能力的抑制作用;2)RT-PCR法检测不同浓度Doxycycline作用48h后,MMP2的mRNA表达情况,同时明胶酶谱法检测MMP2分泌。结果:1)Doxycycline能够以剂量依赖方式抑制大肠癌LS174T细胞的体外运动和侵袭能力,但不影响其粘附能力;2)Doxycycline能够抑制LS174T细胞MMP2的mRNA表达。结论:Doxycycline在体外能够有效地抑制大肠癌LS174T细胞的侵袭、转移,其中MMP2是抑制大肠癌细胞LS174T的一个有效靶点。     

人膀胱癌组织中细胞外基质金属蛋白酶诱导因子的表达

背景与目的:基质金属蛋白酶(matrix metalloproteinases,MMPs)在恶性肿瘤的发生、浸润和转移中发挥关键作用,肿瘤细胞表达的细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,EMMPRIN)司以诱导间质成纤维细胞合成MMPs,从而促进肿瘤的浸润和转移。本研究拟通过检测EMMPRIN在人膀胱癌组织和正常膀胱粘膜中的表达,探讨EMMPRIN在膀胱癌发生、浸润和转移中的作用。方法:应用免疫组化SP方法检测EMMPRIN在45例膀胱癌组织和9例正常膀胱粘膜中的表达,并与临床病理指标结合起来分析。结果;EMMPRIN蛋白定位于癌细胞的胞膜和胞浆,阳性率为73.3％,正常膀胱粘膜和肿瘤间质均为阴性表达。EMMPRIN在Ⅱ、Ⅲ级和术后复发组的阳性表达率分别为84.0％、84.6％和100％,高于Ⅰ级和无复发组(分别为14.3％和55.6％)(P<0.05);在淋巴结转移组的强阳性表达率(85.7％)高于无淋巴结转移组(23.7％)(P<0.05)。结论:EMMPRIN在人膀胱癌组织中过度表达,与膀胱癌的恶性程度有关,可作为判断膀胱 癌是否具备高侵袭转移潜能的指标。