敲减基质金属蛋白酶-10抑制人膀胱癌T24细胞系裸鼠皮下移植瘤生长

目的:探讨敲减MMP-10对人膀胱癌T24细胞系裸鼠皮下移植瘤生长的影响。方法:设计并合成针对人MMP-10的siRNA及阴性对照siRNA,分别注射至裸鼠皮下膀胱癌T24细胞系移植瘤,每周两次监测并比较各组裸鼠体重、瘤体大小,于接种后第5周处死裸鼠,取出皮下移植瘤,并比较各组瘤体重量的差异,并通过免疫组织化学染色检测MMP-10表达水平。结果:皮下注射膀胱癌T24细胞建立裸鼠皮下移植瘤模型,成瘤率达100%。瘤体内注射MMP-10 siRNA可以显著抑制膀胱癌T24细胞裸鼠皮下移植瘤的生长(P<0.05);观察结束时MMP-10 siRNA组所剥离的瘤体重量明显低于对照组(P<0.05)。各组裸鼠的平均体重并没有明显改变(P>0.05),提示瘤体内注射MMP-10 siRNA对裸鼠的一般情况影响不大,并没有明显的毒性或副作用。免疫组织化学染色结果示MMP-10 siRNA组MMP-10的染色强度及阳性细胞数均明显低于普通成瘤组和阴性对照组,提示瘤体内注射siRNA可以在体内有效敲低MMP-10的表达。结论:MMP-10在人膀胱癌T24细胞裸鼠皮下移植瘤的生长过程中发挥重要作用;瘤体内注射MMP-10 siRNA可以有效且安全地抑制膀胱癌T24细胞裸鼠皮下移植瘤的生长。     

MMP-10在人膀胱癌T24细胞系体外侵袭中的作用

目的:探讨基质金属蛋白酶MMP-10在人膀胱癌细胞体外侵袭中的作用.方法:设计并合成针对MMP-10的siRNA和阴性对照siRNA,以脂质体转染法将其分别导入具有体外高侵袭潜能的T24细胞系,RT-PCR和Western blot法检测MMP-10基因及蛋白表达水平.Transwell体外侵袭实验比较干扰MMP-10前后T24细胞体外侵袭潜能的变化.结果:MMP-10-siRNA可有效沉默MMP-10基因的表达,并降低T24细胞系的体外侵袭能力.结论:MMP-10可能在膀胱癌侵袭转移过程中发挥重要作用,抑制MMP-10可为预防及治疗转移性膀胱癌提供新的思路.     

基质金属蛋白酶-10的研究进展

肿瘤浸润和转移在肿瘤的发展过程中起着至关重要的作用.肿瘤的浸润和转移是肿瘤细胞与宿主细胞之间相互作用的连续过程.近年来,介导肿瘤与细胞外基质降解的相关基因引起了人们的极大关注.基质金属蛋白酶(MMPs)则是依赖Zn2 降解细胞外基底膜基质的一类蛋白水解酶.     

胸腺瘤组织中PTEN基因基质金属蛋白酶-2和基质金属蛋白酶-9的表达

目的 探讨PTEN基因、基质金属蛋白酶(MMP)-2和MMP-9在胸腺瘤中的表达及其与胸腺瘤临床病理特征的关系.方法 应用免疫组织化学SP方法,检测45例胸腺瘤和16例非瘤胸腺(对照组)组织中PTEN、MMP-2和MMP-9的表达.结果 PTEN、MMP-2和MMP-9在胸腺瘤组织中的阳性表达率分别为53.5%、48.9%和62.2%,在对照组中的阳性表达率分别为93.8%、6.3%和25.0%,差异均有统计学意义(P<0.05).PTEN和MMP-2的表达与胸腺瘤的恶性程度(世界卫生组织分型)有关(P<0.05),与胸腺瘤的侵袭性(Masaoka分期)有关(P<0.01).MMP-9的表达与胸腺瘤的恶性程度和侵袭性无关(P>0.05).结论 PTEN、MMP-2和MMP-9与胸腺瘤的发生、发展可能有关,且PTEN和MMP-2与胸腺瘤的恶性程度和侵袭性相关.     

基质金属蛋白酶-2在膀胱癌和阴茎癌组织中的表达及其意义

目的　探讨基质金属蛋白酶 (MMP 2 )在膀胱癌和阴茎癌组织中的表达及其意义。方法　采用免疫组化方法 ,检测42例膀胱癌和 45例阴茎癌组织中MMP 2的表达水平。结果　MMP 2在阴茎癌组织中的阳性表达率为 5 7.78% ( 2 6/4 5 ) ,高分化者阳性表达率为 45 .16% ( 14 /3 1) ,中低分化者阳性表达率为 85 .71% ( 12 /14 ) ,2组有显著性差异 (P <0 .0 5 )。MMP 2在膀胱癌组织中阳性表达率为 2 8.5 7% ( 12 /4 2 ) ,在移行细胞癌和腺癌组织中阳性表达率分别为 2 3 .68% ( 9/3 8)和 75 .0 0 % ( 3 /4 ) ,两者比较有非常显著性差异 (P <0 .0 1)。结论　MMP 2水平可作为阴茎癌和膀胱癌术后综合治疗及预后判断的生物学指标     

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。     

血管内皮生长因子和基质金属蛋白酶-2在上皮性膀胱癌组织中的表达

目的：探讨上皮性膀胱癌组织中血管内皮生长因子（VEGF）和基质金属蛋白酶-2（MMP-2）的表达及临床意义。方法采用免疫组化S-P法检测67例上皮性膀胱癌及其癌旁组织中VEGF和MMP-2的表达,并分析两者与患者临床病理特征的关系。结果上皮性膀胱癌组织中VEGF和MMP-2的表达均高于癌旁组织（ P均＜0.05）。上皮性膀胱癌组织中VEGF和MMP-2的表达与其病理分级和临床分期有关（ P均＜0.05）。上皮性膀胱癌组织中VEGF和MMP-2的表达呈正相关关系（ r=0.468,P＜0.05）。结论上皮性膀胱癌组织中VEGF和MMP-2高表达,两者可能参与了上皮性膀胱癌的疾病进展。     

基质金属蛋白酶-2和基质金属蛋白酶-9在直肠癌中的表达及其临床意义

 目的　探讨基质金属蛋白酶-2（MMP-2）及基质金属蛋白酶-9（MMP-9）在直肠癌发病机制及转移中的临床意义。方法　用RT-PCR方法检测MMP-2 mRNA、MMP-9 mRNA在正常肠组织以及直肠癌组织中的表达。结果　MMP-2 mRNA、MMP-9 mRNA在正常肠组织中低表达（0.3150±0.1766、0.3050±0.1995）,而其在有转移的直肠癌患者癌组织中的表达明显增强,与正常对照组比较差异有统计学意义。结论　MMP-2、MMP-9在直肠癌患者中高表达,并且其在癌转移中有重要意义。     

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.     

白桦酯醇对卵巢癌裸鼠皮下移植瘤组织中MMP-2和MMP-9表达的影响及意义

目的 探讨白桦酯醇对卵巢癌SKOV3细胞裸鼠皮下移植瘤组织中基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）表达的影响和意义。方法 建立卵巢癌SKOV3细胞裸鼠移植瘤模型,将30只裸鼠随机分为白桦酯醇高剂量组（80mg／kg）、中剂量组（40mg／kg）、低剂量组（20mg／kg）以及对照组（生理盐水）和紫杉醇组（135mg／m2）,每组6只,腹腔给药02ml/只,隔日1次,连续21d。停药后第2d处死裸鼠,测量其移植瘤体积和瘤重;用免疫组织化学法检测移植瘤组织中MMP-2和MMP-9蛋白的表达并计算表达率。结果 白桦酯醇处理组和紫杉醇组的移植瘤体积和瘤重均小于对照组（P＜0.05）;白桦酯醇中、高剂量组的抑瘤率分别为44.5％和51.4％,均高于其低剂量组的17.5%（P＜0.05）。与对照组比较,白桦酯醇处理组和紫杉醇组移植瘤组织中MMP-2和MMP-9蛋白表达水平均减弱;白桦酯醇低、中、高剂量组移植瘤组织中MMP-2和MMP-9蛋白的表达率（％）分别为47.13±8.67和53.82±7.41、38.29±4.37和48.79±4.63、26.88±5.12和38.93±2.82,均低于对照组的56.78±6.42和69.35±5.03,且白桦酯醇3组之间的差异均有统计学意义（P＜0.05）。结论 白桦酯醇对卵巢癌SKOV3细胞皮下移植瘤的生长有抑制作用,其机制可能与下调MMP-2和MMP-9蛋白的表达有关。     

小窝蛋白-1、基质金属蛋白酶-2在膀胱癌的表达及其关系

Objective: To explore the expression of caveolin-1 (CAV-1) and matrix metalloproteinase-2 (MMP-2) in bladder cancer and its clinical significance. Methods: The expression of CAV-1 and MMP-2 were detected by the SP immunohistochemical method in 77 cases of bladder cancer. Results: The positive rates of CAV-1, MMP-2 in bladder transitional cell carcinoma (BTCC) and bladder adenocarcinoma were significantly high than those of normal bladder mucosa, the positive rates in the deeper cancer invasion was significantly high than those of superficial bladder cancer (P ＜ 0.01), the positive rates of CAV-1 in grades Ⅰ, Ⅱ and Ⅲ of bladder transitional cell carcinaoma (BTCC) were 15%, 40% and 68%, respectively;the positive rates of MMP-2 in grades Ⅰ, Ⅱ and Ⅲ of bladder transitional cell carcinoma (BTCC) were 20%, 40% and 72%,respectively (P ＜ 0.01). CAV-1 was positive associated with MMP-2 in bladder cancer (r= 0.598, P ＜ 0.001), but not in bladder tissue. Conclusion: CAV-1 and MMP-2 were associated with stage and grade of bladder cancer, which suggested that CAV-1might improve secretion of MMP-2.     

Intravesical administration of exogenous microRNA-145 as a therapy for mouse orthotopic human bladder cancer xenograft

We previously reported that the level of microRNA (miR)-145 is attenuated in human bladder cancer cells. In this current study, we investigated whether intravesical administration of miR-145 could be a potential therapeutic strategy for controlling bladder cancer by using an orthotopic human bladder cancer xenograft model. Following transfection of 253J B-V cells with miR-145, the effects of the ectopic expression of miR-145 were examined by performing MTT, Western blotting analysis, Hoechst33342 staining, and wound healing assay in vitro. Also, a mouse orthotopic human bladder cancer model was established by inoculating 253J B-V cells into the bladder wall of mice. The anti-cancer effects of intravesical injections of miR-145 into these mice were then assessed. Transfection of 253J B-V cells with miR-145 induced apoptosis and suppression of cell migration in vitro. Western blotting showed that the levels of c-Myc, socs7, FSCN1, E-cadherin, β-catenin, and catenin δ-1 were decreased and that the PI3K/Akt and Erk1/2 signaling pathways were increased in compensatory fashion. In vivo, mice treated with miR-145 showed 76% inhibition of tumor growth, with a significant prolongation of animal survival (p = 0.0183 vs. control). Western blotting showed that both apoptosis and cell motility-related genes were significantly decreased as seen in vitro. Furthermore, PI3k/Akt and Erk1/2 signaling pathways, which were activated in a compensatory manner in vitro, were decreased in vivo. Intravesical administration of exogenous miR-145 was thus concluded to be a valid therapy for bladder cancer in this human bladder cancer xenograft model.     

The expression of platelet-derived endothelial cell growth factor in human bladder cancer

Platelet-derived endothelial cell growth factor (PD-ECGF) is an angiogenic factor and in some studies PD-ECGF/dTdRPase expression levels in several kind of cancers were higher than in their surrounding normal tissues. In this study, we evaluated PD-ECGF/dTdRPase expression in bladder cancer by an immunohistological method and determined whether it correlated with tumor stage, grade and recurrence. PD-ECGF/dTdRPase expression was correlated with tumor grade and stage. Furthermore, among the superficial tumors, PD-ECGF/dTdRPase expression was correlated with a recurrence-free rate and thus it might be a prognostic factor for the recurrence of superficial bladder cancer.     

Low ANXA10 expression is associated with disease aggressiveness in bladder cancer

Background:

Markers for outcome prediction in bladder cancer are urgently needed. We have previously identified a molecular signature for predicting progression in non-muscle-invasive bladder cancer. ANXA10 was one of the markers included in the signature and we now validated the prognostic relevance of ANXA10 at the protein level.

Methods:

We investigated ANXA10 expression by immunohistochemistry using a tissue microarray with 249 Ta and T1 urothelial carcinomas. The expression of ANXA10 was also investigated in an additional set of 97 more advanced tumours. The functional role of ANXA10 in cell lines was investigated by siRNA-mediated ANXA10 knockdown using wound-healing assays, proliferation assays, and ingenuity pathway analysis.

Results:

Low expression of ANXA10 correlated with shorter progression-free survival in patients with stage Ta and T1 tumours (P<0.00001). Furthermore, patients with more advanced tumours and low ANXA10 expression had an unfavourable prognosis (P<0.00001). We found that ANXA10 siRNA transfected cells grew significantly faster compared with control siRNA transfected cells. Furthermore, a wound-healing assay showed that ANXA10 siRNA transfected cells spread along wound edges faster than control transfected cells.

Conclusion:

We conclude that ANXA10 may be a clinical relevant marker for predicting outcome in both early and advanced stages of bladder cancer.     

膀胱癌中cyclin D1和MMP-2的表达及其临床意义

目的:探讨cyclin D1和MMP-2在膀胱癌中的表达及其与临床病理特征之间的关系。方法:应用逆转录-聚合酶链反应（RT-PCR）方法检测127例膀胱癌和配对癌旁组织以及15例正常膀胱组织中cyclin D1和MMP-2基因的表达,免疫印迹法观测cyclin D1和MMP-2蛋白表达情况。分析两者与膀胱癌临床病理特征之间的关系及两者的相关性。结果:cyclin D1和MMP-2 mRNA在膀胱癌中的表达明显高于癌旁组织和正常膀胱组织。正常膀胱组织中cyclin D1和MMP-2 mRNA的表达率分别为13.3%、20.0%,癌旁组织为26.0%、33.1%,均显著低于癌组织81.1%、88.2%（P<0.05）。类似于基因表达,cyclin D1和MMP-2蛋白在膀胱癌中的表达相对较高。cyclin D1和MMP-2基因在膀胱癌中的表达与病理分级、临床分期和淋巴转移相关(P<0.05),cyclin D1表达还与肿瘤大小有关（P<0.05）,两者均与患者年龄和性别无关（P>0.05)。膀胱癌组织中cyclin D1和MMP-2 mRNA的表达呈正相关(r=0.771,P<0.01)。结论:cyclin D1和MMP-2联合检测可为膀胱癌诊断和治疗提供参考。     

Expression and function role of DNA methyltransferase 1 in human bladder cancer

BACKGROUND:

The identification of potential tumor markers will help improve therapeutic planning and patient management. Therefore, the aim of this study was to highlight the role of DNA methyltransferase 1 (DNMT1) in bladder cancer.

METHODS:

A total of 50 samples of nonmalignant urothelium, 65 of muscle‐invasive bladder cancers, 15 of distant metastasis, and 50 of nonmuscle‐invasive bladder cancers were selected for immunohistochemical staining analysis. Furthermore, human bladder cancer cell lines HT1376 and HT1197 were selected for cell and animal experiments investigating changes in tumor behavior, treatment response, and related signaling in bladder cancer.

RESULTS:

The incidence of nuclear DNMT1 immunoreactivity in the bladder cancer specimens (45%) was significantly higher than in nonmalignant urothelium (15%, P = .0005), and the incidence in cancer was positively linked to clinical stage (24% in ≤T1 vs 55% in T2‐T4, P = .0007). The staining of DNMT1 was also significantly linked to lower complete response rates (P = .0014) and reduced survival rates (P = .000). By in vitro and in vivo experiments, DNMT1 silencing vector reduced tumor growth and attenuated treatment resistance in bladder cancer cells. Less epithelial‐mesenchymal transition, less invasion, and slower tumor growth were noted in cancer cells with inhibited DNMT1. Furthermore, the epidermal growth factor receptor‐mediated phosphatidylinositol 3′–kinase‐protein kinase B pathway might be the mechanism underlying the effects of DNMT1 on bladder cancer.

CONCLUSIONS:

DNMT1 could be a significant clinical predictor for stage and treatment response of bladder cancer. Moreover, targeting this enzyme could be a promising strategy for treating bladder cancer, as evidenced by inhibited tumor growth and enhanced radiosensitivity. Cancer 2011;. © 2011 American Cancer Society.     

The role of narrow-band imaging in the management of non-muscle-invasive bladder cancer

Narrow-band imaging is a young optical enhancement technology for endoscopy. It is a filter to the standard white light which increases the contrast between underlying vasculature and epithelial strata of the mucosa, improving the detection of bladder cancer with particular regard to high grade, flat lesions. Narrow band imaging is absolutely safe, may be used any time during a procedure, either during office cystosopy or transurethral resection, and implies a minimal burden for the healthcare provider given the absence of a learning curve and the limited cost of the camera and light source. The ameliorated detection translates into an improved management of the disease and a lower recurrence risk in prospective randomized studies, suggesting the inclusion of the technology in daily clinical practice.     

靶向MMP-9脱氧核酶抑制人肺腺癌细胞的粘附与迁移的实验研究

背景与目的脱氧核酶(DNAzyme,Deoxyribozyme)具有高效的催化活性和特异的序列识别能力,可特异性切割靶RNA使其失去生物学活性,从而抑制靶RNA的基因表达。本实验通过探讨靶向MMP-9的脱氧核酶对人肺腺癌A549细胞的细胞粘附与迁移能力的影响,为使用脱氧核酶抑制肺癌的浸润转移提供理论依据。方法设计靶向MMP-9的脱氧核酶,应用oligofectamine将脱氧核酶转染到A549细胞中,采用Western blot方法检测细胞中MMP-9的表达,以及利用细胞粘附与迁移试验观察脱氧核酶干预后细胞粘附与迁移能力的变化。结果在靶向MMP-9脱氧核酶干预后,细胞中MMP-9的表达较对照组与空白组显著减低(P<0.01),同时发现细胞的粘附率、迁移速度均显著降低(P<0.01)。结论靶向MMP-9的脱氧核酶能够抑制人肺腺癌A549细胞中MMP-9的表达,并且有效阻止细胞的粘附与迁移。