食管鳞状细胞癌中ADAMTS1mRNA及其蛋白表达

摘 要：［目的］ 评价含凝血酶敏感素基序的去整合素金属蛋白酶1（a disintegrin and metalloproteinase with thrombospondin motifs 1,ADAMTS1）mRNA及其蛋白表达在食管鳞状细胞癌（esophageal squamous cell carcinoma,ESCC）发生发展中的意义。［方法］ 应用RT-PCR方法及免疫组织化学SP方法分别检测ESCC组织及癌旁正常组织中ADAMTS1 mRNA及其蛋白表达的情况。［结果］ ①ADAMTS1mRNA在ESCC组织中表达量显著性低于癌旁正常组织（0.394±0.123 vs 0.895±0.276,P<0.01）;ADAMTS1mRNA在无淋巴结转移组中的表达量显著性低于淋巴结转移组（0.298±0.102 vs 0.482±0.157,P<0.05）。②ESCC组织中ADAMTS1蛋白的阳性表达率显著性低于癌旁组织（38.9% vs 94.4%,P<0.01）。［结论］ ADAMTS1异常表达可能与ESCC的发生、发展及淋巴结转移密切相关。     

食管鳞状细胞癌组织Bin1基因启动子甲基化状态及其临床意义

目的:探讨食管鳞状细胞癌(esophageal squamous cell cancer,ESCC)患者的食管鳞癌组织、癌旁组织中Bin1基因启动子甲基化状态及其mRNA的表达及其临床意义.方法:采用实时荧光定量PCR(qRT-PCR)分别检测58例经病理证实的ESCC患者的食管鳞癌组织、癌旁组织中Bin1基因mRNA的表达情况;用甲基化特异性PCR(MSP)检测上述食管鳞癌组织中Bin1基因启动子甲基化状态,比较ESCC患者Bin1甲基化状态与临床病理分期的关系.结果:ESCC组织中Bin1基因启动子甲基化率明显高于癌旁组织(58.62％ vs 25.86％,x2=12.76,P＜0.01),Bin1甲基化状态与患者TNM分期、肿瘤侵润深度、分化程度、淋巴结转移相关(均P ＜0.05).ESCC组织中Bin1 mRNA的表达水平明显低于癌旁组织[(0.78 ±0.05) vs (1.03±0.03),t=9.643,P＜0.01)];发生Bin1甲基化的组织中Bin1 mRNA表达水平明显低于未发生甲基化的组织[(0.68±0.04) vs (0.85±0.07),t=2.476,P＜0.05].结论:Bin1基因启动子区甲基化状态可能与ESCC的发生密切相关,它是ESCC中Bin1 mRNA低表达或缺失的机制之一,且与ESCC进展和淋巴结转移有关.     

DNA启动子甲基化致ADAMTS9蛋白下调促进大肠癌病程进展

背景与目的：大肠癌的发病率逐年递增。该研究旨在考察ADAMTS9蛋白水平及其启动子甲基化水平在大肠癌发病及病程进展中的意义。方法：采用甲基化特异性PCR法,检测162例大肠癌患者外周血来源DNA样本中ADAMTS9基因启动子甲基化水平;并应用酶联免疫吸附试验法检测162例大肠癌患者和150例健康体检者血浆ADAMTS9蛋白水平。结果：与健康人相比,大肠癌患者血浆中ADAMTS9蛋白水平明显降低[(65.25±9.70)μg vs (50.28±9.66)μg,P<0.001];162例大肠癌患者中有66例ADAMTS9基因启动子存在甲基化(40.7%);甲基化患者的血浆ADAMTS9蛋白水平显著降低(P<0.001),而ADAMTS9蛋白低表达患者的血样ADAMTS9基因甲基化水平显著升高(P=0.007);ADAMTS9甲基化与肿瘤大小(P=0.017)和肿瘤分化程度(P=0.029)密切相关,而ADAMTS9蛋白低表达与浸润深度(P=0.020)、淋巴结转移(P=0.019)和Dukes分期(P=0.002)密切相关。结论：在大肠癌中,由DNA启动子甲基化引起的ADAMTS9蛋白表达下调可能参与大肠癌的发病、侵袭转移、并促进病程进展。     

GRHL3和c-Myc在食管鳞状细胞癌组织中的表达及其临床意义

目的:研究食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)患者肿瘤组织及癌旁组织中GRHL3和c-Myc的表达情况及其临床意义.方法:收集2015年4月至2016年7月在河北医科大学第四医院胸外科行肿瘤切除并经病理证实的64例ESCC患者的肿瘤组织、癌旁组织,采用Real-time PCR法和免疫组化法检测GRHL3和c-Myc基因的mRNA和蛋白表达情况,分析其与患者临床特征的关系.结果:与癌旁组织相比,ESCC组织中GRHL3 mRNA表达水平和蛋白阳性表达水平均显著升高[(2.85±2.83) vs(2.06±2.02),P＜0.01;81.30％ vs 25.00％,P＜0.01],ESCC组织中c-Myc mRNA表达水平和蛋白阳性表达水平均显著升高[5.13±5.11)vs (2.03±2.00),P＜0.01;42.20％ vs.20.30％,P＜0.01].ESCC组织中GRHL3 mRNA的表达与c-Myc mRNA的表达呈显著正相关关系(P＜0.05),GRHL3蛋白表达和c-Myc蛋白表达也呈显著正相关(P＜0.01).GRHL3蛋白表达和c-Myc蛋白表达与患者肿瘤浸润程度、淋巴结转移、临床分期、分化程度相关.结论:ESCC患者肿瘤组织中GRHL3与c-Myc表达水平显著提高,两者表达呈正相关,且两者与患者临床病理特征密切相关,可能是影响ESCC病理进程的重要因素.     

ADAMTS9基因启动子甲基化在直肠癌中的表达与临床病理特征的相关性

目的检测直肠癌组织中ADAMTS9基因转录和启动子甲基化状态,探讨其临床意义。方法收集2016年1月至2018年12月间于延安大学附属医院普通外科医院行直肠癌根治手术切除的83例直肠癌组织及其相应的癌旁组织标本,通过实时定量PCR和巢式-甲基化特异性PCR检测ADAMTS9基因转录和启动子甲基化状态,分析ADAMTS9基因启动子甲基化与直肠癌临床病理特征的相关性。结果直肠癌组织中ADAMTS9 mRNA表达水平低于配对癌旁组织(χ²=1161,P<0.0001)。83例直肠癌组织中ADAMTS9基因甲基化发生率20.5%,高于癌旁组织7.2%(6/83)(χ²=37.19,P<0.0001)。ADAMTS9基因启动子区异常甲基化增加直肠癌发病风险(OR=10.254,95%CI=3.395~25.631)。直肠癌组织中ADAMTS9启动子区甲基化与肿瘤TNM分期、分化程度相关。结论ADAMTS9基因启动子异常甲基化参与了直肠癌的发生发展过程。     

贲门腺癌组织中Smad基因家族不同成员的表达及其临床意义

目的:检测贲门腺癌(gastric cardia adenocarcinoma,GCA)中Smad基因家族不同成员的表达水平及其相关性,并探讨其临床意义。方法:收集河北医科大学第四医院2004至2009年间病理确证的贲门腺癌患者110例,RT-PCR检测贲门腺癌和癌旁组织中Smad2、Smad3、Smad4和Smad7 mRNA的表达,免疫组织化学方法检测贲门腺癌组织和癌旁组织中p-Smad2/3、Smad4和Smad7蛋白的表达并分析其相关性,分析p-Smad2/3、Smad4和Smad7蛋白表达与贲门腺癌临床病理特征的关系。结果:贲门腺癌组织中Smad2、Smad3和Smad4 mRNA表达水平均显著低于相应癌旁组织[(0.4956±0.1862)vs(0.8611±0.2914),P<0.01;(0.4713±0.1712)vs(0.8314±0.2811),P<0.01;(0.5145±0.1987)vs(0.8954±0.2856),P<0.01],而Smad7 mRNA表达水平显著高于癌旁组织[(0.5114±0.1962)vs(0.2012±0.1006),P<0.01]。贲门腺癌组织p-Smad2/3、Smad4蛋白表达的阳性率显著低于癌旁组织(42.7%vs 93.6%,P<0.01;45.5%vs 95.5%,P<0.01),且与肿瘤TNM分期和组织分化程度密切相关(P<0.05)。贲门腺癌组织Smad7蛋白表达阳性率显著高于癌旁组织(48.2%vs 3.6%,P<0.01),且与肿瘤组织分化程度密切相关(P<0.05)。贲门腺癌中p-Smad2/3与Smad4蛋白的表达呈正相关,但它们均与Smad7蛋白表达无明显的相关性。结论:贲门腺癌组织低表达Smad2、Smad3和Smad4,高表达Smad7,Smad基因的异常表达可能参与贲门腺癌的发生、发展。     

食管鳞状细胞癌中ERK的表达及其临床意义

目的:研究食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)及癌旁黏膜组织中的细胞外信号调节激酶(extracellular signal ragulated kinase,ERK)mRNA的表达,探讨ERK在ESCC发生、发展中的作用及其意义。方法:收集河北医科大学第四医院胸外科40例ESCC患者癌组织标本及25例患者的癌旁组织标本,RT-PCR检测ESCC与癌旁组织中ERK mRNA的表达情况,分析其与ESCC临床病理特征的关系。结果:ESCC组织中ERK mRNA的表达明显增高,而在癌旁组织中呈现低表达(0.656±0.055 vs 0.450±0.070,P<0.01)。ESCC组织中ERK mRNA的表达与ESCC患者的年龄、性别和组织分化程度无关(P>0.05),而与临床分期、淋巴结转移显著相关(P<0.05)。结论:ESCC组织高表达ERK mRNA,ERK可能在ESCC发生、发展中起重要的作用。     

食管鳞癌组织和引流淋巴结中IDO和BIN1的表达及其临床意义

目的:探讨食管鳞癌(esophageal squamous cell carcinoma,ESCC)患者肿瘤组织和引流淋巴结(tumor draininglymph node,TDLN)组织吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)和桥接整合因子1(bridging integrator 1,BIN1)的表达及其在ESCC进展中的意义.方法:收集2013年4月至2014年7月在河北医科大学第四医院胸外科行肿瘤切除并经病理证实的71例ESCC患者的肿瘤组织、癌旁组织和TDLN标本,其中肿瘤组织以癌旁组织作为对照组,转移淋巴结以未转移淋巴结作为对照组,均采用Real-time PCR法和免疫组化法检测IDO和BIN1表达情况,分析两者表达的相关性及其与患者临床特征的关系.结果:与未转移淋巴结相比,转移淋巴结中IDO mRNA表达水平和蛋白阳性表达率均显著增高(0.47±0.14vs0.22 ±0.09,P＜0.01;90.91％ vs 67.35％,P=0.042),而BIN1 mRNA表达水平和蛋白阳性表达率均显著降低(0.15 ±0.11 vs 0.35 ±0.15,P＜0.01;50.00％ vs 77.55％,P=0.028).与癌旁组织相比,肿瘤组织中IDO mRNA表达水平和蛋白阳性表达率均显著增高(0.51 ±0.12 vs 0.24 ±0.11,P＜0.01;81.69％ vs 22.54％,P＜0.01),而BIN1 mRNA表达水平和蛋白阳性表达率均显著降低(0.17±0.10 vs0.41±0.14,P＜0.01;19.72％ vs 80.28％,P=0.006).肿瘤组织和TDLN中,IDO蛋白表达与肿瘤侵犯范围、淋巴结转移、临床分期相关,而BIN1蛋白表达与肿瘤分化程度、侵犯范围、淋巴结转移、临床分期相关.结论:ESCC患者肿瘤组织和转移TDLN中IDO表达水平显著提高、BIN1表达水平显著降低与患者临床特征密切相关,可能是影响ESCC进展的重要因素.     

PAR1 mRNA及蛋白在食管鳞状细胞癌中的表达及临床意义

[目的]探讨PAR1 mRNA及蛋白在食管鳞状细胞癌及其匹配癌旁组织中的表达和临床意义。[方法]应用RT-PCR方法检测35例食管鳞状细胞癌患者食管原位癌组织及其匹配癌旁组织PAR1 mRNA的表达水平,并分析其与患者性别、年龄、肿瘤大小、临床分期及是否淋巴结转移等临床指标的关系;应用免疫组织化学法检测55对食管鳞癌组织及其匹配癌旁组织的PAR1蛋白表达情况。[结果]食管鳞癌组织PAR1 mRNA的表达水平显著高于其匹配的癌旁组织(146.220±24.790vs134.054±23.593,t=-4.17,P<0.01),而且与淋巴结是否转移有关(t=-2.199,P<0.05);食管鳞癌组织PAR1蛋白表达阳性率显著高于其匹配的癌旁组织(49%vs3.64%,χ2=23.04,P<0.001)。[结论]PAR1 mRNA及蛋白在食管鳞癌中的表达均显著上调,可能参与食管癌的发生发展、侵袭与转移。     

胃癌和转移淋巴结组织ADAMTS1表达及其生物学意义的探讨

目的:探讨含(I)型血小板结合蛋白基序的解聚蛋白样金属蛋白酶( ADAMTS1)在胃癌及转移淋巴结组织中的表达,分析其与胃癌浸润深度、分化程度及淋巴结转移等临床病理学参数的关系.方法:免疫组化法检测72例成对胃癌组织、癌旁组织及胃周淋巴结组织标本ADAMTS1的表达,半定量评分系统评估染色,分析ADAMTS1表达与胃癌临床病理学参数的关系.结果:ADAMTS1在胃癌组织中弱阳性率、阳性率及强阳性率分别为37.5％(27/72)、4.2％(3/72)和1.4％(1/72),在癌旁组织中分别为58.3％(42/72)、25.0％(18/72)和11.1％(8/72),在转移淋巴结组织中分别为55.9％(33/59)、20.3％(12/59)和3.4％(2/59),在无转移淋巴结组织中分别为46.2％(6/13)、0(0/13)和0(0/13).胃癌组织ADAMTS1表达明显低于癌旁组织,z=-6.481,P＜0.000 1;转移淋巴结组织ADAMTS1表达明显高于胃癌组织(z=4.164,P＜0.000 1)和无转移淋巴结组织(z=2.130,P=0.033).ADAMTS1在有淋巴结转移的胃癌组织中表达明显高于无淋巴结转移的胃癌组织,z=2.045,P=0.041.结论:ADAMTS1在胃癌原发灶中呈现低表达,在转移淋巴结中呈现高表达.ADAMTS1在胃癌中表达与淋巴结转移有关,表明ADAMTS1基因表达在胃癌原发肿瘤生长及转移中可能存在不同的作用机制,可能与肿瘤微环境有关.     

ADAMTS8 and ADAMTS15 expression predicts survival in human breast carcinoma

We recently undertook expression profiling of all 19 human ADAMTS metalloproteinases (a disintegrin and metalloproteinase with thrombospondin motifs) in malignant and non-neoplastic breast tissue and showed that 11 of the ADAMTS genes are dysregulated in breast carcinoma. We identified a subgroup of ADAMTSs, based on functional and amino acid sequence similarity (ADAMTS1, 4, 5, 8 and 15), to be the focus of further study in breast carcinoma. Further RNA expression analysis by real-time PCR on a different cohort of 229 patients with breast cancer has identified ADAMTS8 as a predictor of poor overall survival (OS) (hazard ratio (HR) = 2.20, 95% C.I. = 1.29-3.74, p = 0.004) and confirmed ADAMTS15 as a predictor of prolonged relapse-free survival (RFS) (HR = 0.54, 95% C.I. = 0.32-0.89, p = 0.016). We explored the differences in survival of the 4 groups that could be categorized based on the expression levels of ADAMTS8 and ADAMTS15. For both RFS and OS, the group with high ADAMTS8 and low ADAMTS15 expression had a particularly poor prognosis. This group had a 3-fold higher chance of recurrence (HR = 3.03, 95% C.I. = 1.49-6.15, p = 0.001) and a greater than 5-fold higher chance of death (HR = 5.40, 95% C.I. = 2.16-13.5, p < 0.001) than the most favorable prognostic group. This prediction of poor prognosis by ADAMTS8 and ADAMTS15 expression was found to be independent of other classical clinicopathological factors. Results observed in FVB-PyMT mice, a robust transgenic model of highly metastatic breast carcinoma, fitted the expectation that relatively high expression levels of ADAMTS8 together with low expression levels of ADAMTS15 seen in human breast carcinoma are associated with a poor clinical outcome. In summary, ADAMTS8 and ADAMTS15 have emerged as novel predictors of survival in patients with breast carcinoma.     

Matrix-degrading proteases ADAMTS4 and ADAMTS5 (disintegrins and metalloproteinases with thrombospondin motifs 4 and 5) are expressed in human glioblastomas

Brain tumors, in particular glioblastomas, have a high morbidity and mortality, mainly due to their invasive nature. A prerequisite for this invasiveness is cell migration based on increased expression of proteases digesting the extracellular matrix. Brevican, an important extracellular proteoglycan that is upregulated in glioblastomas, can be degraded by certain proteases. We demonstrate that in human glioblastomas secretory proteases like ADAMTS4 and ADAMTS5 (aggrecanases 1 and 2; ADAMTS = a disintegrin and metalloproteinase with thrombospondin motifs) are expressed on the mRNA and protein levels in considerable amounts. Real-time RT-PCR shows a higher levels of ADAMTS4 and 5 expressions in glioblastomas in situ, compared to cultured human glioblastoma cells. The upregulation of these proteases in vivo by cytokines may explain this difference. In vitro, transforming growth factor-beta induces ADAMTS4, but less ADAMTS5, and interleukin-1beta ADAMTS5, but not ADAMTS4. As demonstrated by immunohistochemistry and confocal microscopy in situ, ADAMTS5 expression is confined to proliferating glioblastoma cells of surgical tumor sections and with lower intensity to astroglial cells in normal brain sections, as opposed to brevican. In vitro, glioblastoma-derived ADAMTS5 degrades recombinant human brevican to several smaller fragments. Our results show that ADAMTS4 and 5 are upregulated on proliferating glioblastoma cells and these proteases may contribute to their invasive potential.     

ADAMTS4 and its proteolytic fragments differentially affect melanoma growth and angiogenesis in mice

The metalloproteinase ADAMTS4 (ADAMTS, a disintegrin‐like and metalloproteinase with thrombospondin motif)/aggrecanase‐1 is highly expressed in cartilage and has been implicated in human arthritis. Although abundantly expressed in many types of cancer, its role in cancer remains unknown. In this work, we demonstrate for the first time that full‐length ADAMTS4 and its catalytically more active N‐terminal 53 kDa autocatalytic fragment both promote B16 melanoma growth and angiogenesis in mice. In contrast, overexpression of its catalytically inactive E362A mutant or truncated fragments containing only the C‐terminal ancillary domains suppresses melanoma growth and angiogenesis under similar conditions. Structure–function mapping revealed that the single thrombospondin‐type 1 repeat domain is essential and sufficient for the antitumorigenic activity displayed by the catalytically inactive ADAMTS4 isoforms. Suppression of tumor growth and angiogenesis in mice is accompanied by a significant increase in tumor cell apoptosis, whereas tumor cell proliferation is not affected. Importantly, we identified and demonstrated the presence of novel proteolytic fragments of ADAMTS4 containing essentially only the C‐terminal ancillary domains in cultured cells, and also in human cancer tissues, coexisting with full‐length and catalytically active N‐terminal fragments. The contrasting functions toward tumor growth in mice by the wild‐type proteinase and its catalytically inactive mutant correlate with their contrasting influences on angiogenesis signaling pathway molecules in B16 melanoma in mice. Our results suggest a complex role for ADAMTS4 in cancer with the functional balance of protumorigenic and antitumorigenic isoforms likely to act as an important parameter in determining the net influence of this metalloproteinase on tumor growth in vivo.     

ADAMTS1 as potential prognostic biomarker promotes malignant invasion of glioma

International Journal of Clinical Oncology - Glioma is the most common intracranial malignancy in adults with a high degree of malignancy and poor prognosis, which is largely attributed to the...     

Thrombotic thrombocytopenic purpura: a thrombotic disorder caused by ADAMTS13 deficiency

A serious disorder with characteristic microvascular thrombosis involving the brain and other organs, thrombotic thrombocytopenic purpura (TTP) typically presents with thrombocytopenia, hemolysis with schistocytes on blood smears, and mental changes or seizures. It may progress rapidly to a fatal end if the patient is not treated immediately with plasma. Recent advances have shown that TTP is caused by deficiency of a circulating, von Willebrand factor cleaving metalloprotease, ADAMTS13. This new knowledge will provide clues to improve the diagnosis and management of this intriguing disease.     

Relevance of IGFBP2 proteolysis in glioma and contribution of the extracellular protease ADAMTS1

Expression of IGFBP2 (Insulin-like Growth Factor Binding Protein 2) has been positively correlated with glioma progression. Although the proteolysis of IGFBP2 has been widely recognized, with consequences as a major modulator of IGFII signaling, the relevance of this post-translational modification has not been well studied in tumors. Using an in vivo proteomic approach by Isotope-Coded Protein Label (ICPL), we identified IGFBP2 as a target of the extracellular protease ADAMTS1 (A Disintegrin And Metalloproteinase with ThromboSpondin motifs 1). Notably, the proteolytic pattern of IGFBP2 was also detected in human glioma culture cells and, more importantly, in all glioma samples evaluated. In addition, high expression of ADAMTS1 correlates with higher levels of cleaved IGFBP2 in glioblastoma multiforme cases. Using gene expression public databases, we confirmed that IGFBP2 is a poor prognosis marker for gliomas, and we also observed an important contribution of ADAMTS1.Finally, we showed the impact of ADAMTS1 on IGFII-mediated IGF1R phosphorylation and cellular migration. Our results support a functional interaction between IGFBP2 and ADAMTS1 and suggest the need to evaluate post-translational modifications of IGFBP2 in glioma, in order to approach new therapies.     

High-resolution melting analysis of ADAMTS18 methylation levels in gastric, colorectal and pancreatic cancers

A disintegrin and metalloprotease with trombospondin motifs (ADAMTS) is a family of proteins characterized by the presence of a metalloproteinase domain linked to a variety of specialized ancillary domains. ADAMTS18 is a putative tumor suppressive gene related to nasopharyngeal carcinoma. We used high-resolution melting (HRM) analysis to detect the methylation levels of ADAMTS18 gene in 100 gastric cancers, 100 colorectal cancers, 70 pancreatic cancers, and equal number of adjacent normal tissues. The frequency of ADAMTS18 methylation in all three types of cancers was significantly higher than that in normal tissues. Expression levels of ADAMTS18 were inversely correlated with methylation levels. No significant association was found between ADAMTS18 methylation status and TNM staging in the cancers. In summary, epigenetic regulation of ADAMTS18 was associated with carcinogenesis. The application of HRM analysis is a fast and high-throughput way to investigate the epigenetic status of ADAMTS18.     

Overexpression of ADAMTS5 can regulate the migration and invasion of non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is the major cause of cancer-related lethality among human cancer patients globally, and the poor prognosis of this cancer is mainly explained by metastasis, so it is essential to find out the molecule mechanisms and a novel therapeutic for NSCLC. A disintegrin and metalloprotease with thrombospondin motif 5 (ADAMTS5) belongs to the protease family. It has been reported to participate in tumor migration and invasion. In this study, we showed that the expression of ADAMTS5 was higher in lung cancer tissues by Western blot. The immunohistochemistry analysis was performed in 140 NSCLC cases, and the result indicated that ADAMTS5 was significantly associated with clinical pathologic variables. The Kaplan–Meier curve showed that the high expression of ADAMTS5 was related to poor prognosis of lung cancer patients. Wound healing assays and transwell migration assays revealed that the high expression of ADAMTS5 promoted the migration and invasion of NSCLC. In a word, our findings suggest that ADAMTS5 can regulate the migration and invasion of NSCLC and it may be a useful target of therapy in NSCLC.