RECK、MMP 9在三阴性乳腺癌组织芯片中的表达及临床意义

【摘要】 目的 探讨基质金属蛋白酶抑制剂(RECK)、基质金属蛋白酶 9(MMP 9)在三阴性乳腺癌（TNBC）组织、癌旁组织中的表达及与TNBC临床病理的关系。方法 选择绵阳市中心医院乳腺外科2011年5月~2013年7月经病理证实为TNBC的72例石蜡标本及同 患者癌旁组织,利用免疫组化组织芯片技术检测其RECK、MMP 9蛋白的表达。结果 RECK在TNBC组织及癌旁组织中阳性表达率分别为4306%、8056%（P=0000）。TNBC中RECK的表达与临床分期（P=0009）、组织学分级（P=0010）、腋窝淋巴结转移情况（P=0000）相关。MMP 9在TNBC组织及癌旁组织中阳性表达率分别为625%、1528%（P=0000）。TNBC中MMP 9的表达与肿瘤大小（P=0017）、临床分期（P=0001）、组织学分级（P=0001）、腋窝淋巴结转移状况（P=0001）、Ki 67表达情况（P=0034）相关。TNBC中RECK与MMP 9表达呈负相关(r= 0195,P＜005)。结论 RECK的表达缺失与MMP 9过度表达与TNBC浸润、转移有关,有望成为TNBC的预后指标,并且有可能成为TNBC治疗的靶点。     

Comparison of manual and digital microvascular density counting of RECK expression in glioma

A microvascular density (MVD) counting method for reversion‐inducing cysteine‐rich protein with Kazal motifs (RECK) expression, using a digital image analysis tool, has advantages over manual counting by microscope. Thirty glioma cases with RECK staining were photographed at a magnification of 200× high power field and the photographs in RGB images were analyzed, and stained vessels were captured and were counted automatically. MVD with RECK expression using a digital image analysis tool showed comparable results to those of the manual method. RECK intensity expression could show linear correlation with grades of glioma by the digital method, which was superior compared to the manual method. The present method is recommended to researchers undertaking MVD study for glioma.     

The clinicopathologic relevance of RECK gene polymorphisms in ameloblastoma

ObjectiveTo investigate the relationship between RECK gene polymorphisms and the clinicopathologic features of ameloblastoma.DesignNormal gingival mucosa specimens were obtained from 10 healthy volunteers. Ameloblastomas were surgically removed from 30 patients and part of the tumor specimens were used to detect RECK gene polymorphisms by using PCR-single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing analysis. Expression of RECK and MMP-9 protein was measured using western blot.ResultsThe overall SNP rate was 46.7% (14/30). Four polymorphisms were detected in exon 9, 11, 13, 15 of the RECK gene: two synonymous (P520P and R625R) and two missense SNPs (V275I and I395 V). RECK protein expression in specimens with minor RECK SNPs was lower than that in specimens without RECK SNPs (P < 0.05), and, RECK protein expression in specimens with and without RECK SNPs was lower than that in the normal gingiva specimens (P < 0.05). MMP-9 protein expression in specimens with minor RECK SNPs was higher than that in specimens without RECK SNPs (P < 0.05), and MMP-9 protein expression in specimens with and without RECK SNPs was higher than that in normal gingiva specimens (P < 0.05). RECK gene polymorphisms were closely associated with active proliferation, capsular invasion, and clinical recurrence of ameloblastoma.ConclusionThe rs16932912(G/A) SNP in the RECK gene was closely associated with active proliferation, capsular invasion, and clinical recurrence of ameloblastoma. RECK protein expression was closely associated with the presence of the rs16932912(G/A) SNP.     

RECK、MMP-2在牙周炎患者牙龈组织中的表达

目的:研究RECK蛋白和MMP-2在牙周炎中的表达及其相互关系。方法:应用免疫组化SP法检测56例牙周炎患者牙周组织和16例正常牙龈组织中RECK和MMP-2的表达。结果:RECK在16例正常牙龈组织中的阳性表达率100%,而在牙周炎牙龈组织中的阳性表达率为51.8%。MMP-2在正常牙龈组织中的阳性表达率为31.3%,而在牙周炎牙龈组织中的阳性表达率为76.8%。牙周炎中RECK与MMP-2的表达呈负相关(P<0.01,r=-0.300)。结论:RECK可能通过调控MMP-2来影响牙周炎的发生与发展。     

Matrix metalloproteinase-9 activity is associated with poor prognosis in T3-T4 node-negative colorectal cancer

Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) is a novel membrane-anchored matrix metalloproteinase (MMP) inhibitor. RECK, MMP-2, and MMP-9 are believed to play crucial roles in tumor progression. This study was designed to examine the prognostic value of RECK, MMP-2, and MMP-9 in conjunction with other clinicopathologic factors in patients of T3-T4 node-negative colorectal cancer. RECK and MMP expression was observed using immunohistochemical analysis of the primary tumor from 89 patients with curatively resected T3-4 N0 colorectal cancer retrospectively. High RECK expression was observed in 51 cases, whereas expression was low in the other 38 cases. MMP-2 and MMP-9 expression was positive in cancer cells in 24 and 33 cases, respectively. RECK and MMP-2 expression was not significantly associated with any clinicopathologic factors. However, expression of MMP-9 was correlated with tumor location. A statistically significant inverse correlation was found between RECK and MMP-2 expression, and a statistically significant correlation was found between MMP-2 and MMP-9 expression. However, no association between RECK and MMP-9 expression was observed. Univariate analysis demonstrated that rectal tumor location, preoperative carcinoembryonic antigen more than 5 ng/mL, positive lymphatic invasion, less than 12 dissected lymph nodes, and positive MMP-9 expression were poor prognostic factors of disease-free survival. A multivariate analysis confirmed that enhanced MMP-9 expression was an independent and significant factor for prediction of a poor prognosis. In addition, positive lymphatic invasion and less than 12 dissected lymph nodes were significant negative prognostic factors. In conclusion, MMP-9 status represents a novel prognostic factor in evaluation of T3-T4 node-negative colorectal cancer.     

ERK1/2信号通路阻断剂PD98059对胃癌细胞MGC-803中RECK、MMP-9基因表达的影响

目的:探讨ERK1/2信号通路在RECK基因抑制侵袭转移中的作用.方法:体外培养人胃癌MGC-803细胞,采用MTT法观察胃癌细胞MGC-803增殖情况,Western blot检测P-ERK的蛋白表达,RT-PCR法和Western blot法检测RECK和基质金属蛋白酶9(MMP-9) mRNA和蛋白表达.结果:M...     

RECK蛋白与微血管密度在外耳道及中耳鳞癌中的表达及其相关性

目的:检测RECK蛋白与微血管密度(MVD)在外耳道及中耳鳞癌中的表达并研究其临床意义.方法:采用免疫组化SP法检测26例外耳道及中耳鳞癌组织中RECK与CD34标记的MVD.结果:RECK在外耳道及中耳鳞癌中阳性率为42.3%,在外耳道皮肤中阳性率为80.0%,两组比较差异有统计学意义(P<0.05);外耳道及中耳鳞癌中MVD为(33.58±3.04)个/HP,外耳道皮肤中为(22.50±5.22)个/HP,两组比较差异有统计学意义(P<0.01).外耳道及中耳鳞癌中RECK的表达和MVD、组织学分级及肿瘤分期密切相关.RECK和CD34的表达呈负相关(P<0.05).结论:RECK和CD34的表达呈负相关,二者可能参与了肿瘤的发生,在外耳道及中耳鳞癌的侵袭和转移中发挥重要作用,二者联合检测可望成为外耳道及中耳鳞癌早期诊断和预后判断的分子指标之一.     

抑制miRNA-21表达对胆管癌细胞凋亡、侵袭的影响及其靶基因探讨

目的 探讨抑制miRNA-21表达对胆管癌细胞株凋亡、侵袭的影响及其可能调控的靶基因。方法通过RNAi技术抑制胆管癌细胞株RBE中miRNA-21的表达并加以验证。通过荧光定量PCR技术及蛋白质免疫印迹法(Western blot)检测RBE细胞中RECK基因表达的改变。应用流式细胞仪检测细胞凋亡的变化。用体外侵袭实验分析胆管癌细胞侵袭能力的改变。结果miRNA-21在胆管癌细胞株RBE中的表达得到有效抑制,并且RECK基因及其蛋白的表达明显上调。流式细胞仪显示RBE细胞凋亡率明显增高,且抑制miRNA-21的表达后胆管癌细胞的侵袭性明显下降。结论抑制miRNA-21的表达可促进RBE细胞的凋亡,并降低其侵袭能力,RECK作为miRNA-21的靶基因参与调控过程。     

基质金属蛋白酶抑制剂RECK基因在前列腺细胞株中的表达及意义

目的:探讨RECK基因及基质金属蛋白酶-9(MMP-9)mRNA在不同前列腺细胞株BPH-1、DU-145、LNCaP及PC-3中的表达差异及其意义。方法:应用RT-PCR检测4种不同前列腺细胞株(BPH-1、DU-145、LNCaP及PC-3)中RECK及MMP-9 mRNA的表达;应用W estern印迹法检测不同前列腺细胞株中RECK蛋白的表达。结果:前列腺癌细胞株DU-145、LNCaP及PC-3中RECKmRNA的表达较良性前列腺增生细胞株BPH-1中明显降低(P<0.01),MMP-9 mRNA表达则明显升高。DU-145、LNCaP及PC-3细胞株中RECK蛋白的表达较BPH-1的表达明显降低。结论:RECK基因在前列腺癌中的表达量明显下降,而MMP-9的表达量明显升高,RECK基因可能是一种抑癌基因,其抑癌作用可能与其抑制MMP-9的表达有关。     

RECK和VEGF-C在胃癌组织中的表达和临床意义

目的：研究RECK（reversion-inducing cysteine-rich protein with Kazal motifs）和血管内皮生长因子-C（vascular endotnelial growth factor C,VEGF-C）在胃癌及正常胃组织中的表达,探讨二者在胃癌的发生、发展、浸润和转移中的作用。方法：选择临床病理资料齐全的胃癌蜡块标本158例为实验组,另取正常胃黏膜标本15例为对照组。采用SP免疫组织化学方法检测RECK、VEGF-C在其中的表达并进行χ2检验和相关分析。结果：胃癌组织中RECK呈低表达,且肿瘤浸润深、有淋巴转移、临床分期高、病理分化低时降低;VEGF-C呈高表达,且肿瘤直径大、肿瘤浸润深、有淋巴转移、临床分期高、病理分化低时增高。胃癌中RECK与VEGF-C的表达呈负相关。结论：胃癌组织中RECK呈低表达,VEGF-C呈高表达,检测二者的表达对胃癌的诊断、防治和预后的判断有指导意义。