乙酰肝素酶mRNA在大肠癌中的表达及意义

目的:探讨乙酰肝素酶在大肠癌中的表达及意义.方法:应用原位杂交方法检测乙酰肝素酶mRNA在45例大肠癌组织中的定位及表达.结果:45例大肠癌组织中,乙酰肝素酶mRNA阳性表达22例(48.88%).26例大肠癌伴淋巴结转移者,其原发灶内乙酰肝素酶mRNA表达明显高于无转移组,有显著差异(P＜0.01).乙酰肝素酶mRNA表达与大肠癌组织浸润深度、淋巴结转移有关.结论:乙酰肝素酶可能在大肠癌的生长、浸润和转移中起一定的作用.     

宫颈癌中HPA的表达与细胞增殖、血管生成和转移的关系

目的通过对宫颈癌组织中乙酰肝素酶(heparanase,HPA)、细胞核增殖相关抗原Ki-67和微血管密度的检测,探讨HPA在宫颈癌发生发展中的作用。方法采用免疫组化染色(S-P法)检测67例宫颈癌中HPA、Ki-67和CD34的表达,并与13例正常宫颈组织进行对照研究,分析HPA与患者临床特征、Ki-67和CD34之间的关系。结果67例宫颈癌中49例(73%)阳性表达,而正常13例宫颈组织中无一例阳性表达(P=0.000)。HPA与临床分期、淋巴结转移、细胞增殖和血管生成显著正相关(P值分别为0.001、0.012、0.000、0.000)。结论HPA可能在宫颈癌的发展、浸润、转移和血管生成中起重要作用,并与肿瘤细胞的增殖活性有关,可作为临床预测宫颈癌浸润转移的一个重要参考指标和有价值的治疗靶点。     

Normal colon tissue and colon carcinoma show no difference in heparanase promoter methylation

Introduction

Heparanase, the sole heparan sulfate degrading enzyme, has a role in cellular invasion. Accordingly, a large number of studies have demonstrated an association between heparanase expression and tumor stage and patients' prognosis. In colon carcinoma, heparanase shows increased expression in tumor compared to normal tissue and its expression correlates with the presence of metastasis. One of the regulatory mechanisms of heparanase expression is de-methylation on its promoter. In the present study we evaluated the role of heparanase promoter methylation in colon carcinoma.

Material and methods

Analysis of heparanase promoter methylation was done on 32 samples of colon carcinoma as well as 30 samples of normal colonic mucosa. DNA was extracted from FFPE tissue and subjected to bisulfite conversion. The relative fraction of methylated and unmethylated DNA was evaluated using quantitative real-time PCR.

Results

The fraction of methylated DNA was 1 ± 3.4% in the colon carcinoma group, and 2.5 ± 3.3% in the normal colon group (P = 0.11). Only one case in the normal group and one case in the tumor group showed more than 10% methylation in the heparanase promoter.

Conclusion

We did not find any significant difference in heparanase promoter methylation between colon carcinoma and normal colonic mucosa, suggesting that heparanase overexpression in colon carcinoma is mediated by other mechanisms.     

Salmonella alters heparanase expression and reduces tumor metastasis

Salmonella causes salmonellosis, is a facultative anaerobe and is one of the common Gram-negative bacteria. Salmonella has anti-tumor potential and tumor-targeting activity. The heparin sulfate on cell surfaces can be cleaved by heparanase that is an endo-β-D-glucuronidase. Heparanase can destroy the extracellular matrix and is involved in tumor metastasis and angiogenic activity. Previously, Salmonella was demonstrated to inhibit tumor metastasis. It remains unclear whether Salmonella inhibits metastasis by regulating heparanase. The expression of heparanase in Salmonella-treated tumor cells was found to be decreased. Transwell and wound-healing assays demonstrated the inhibition of cell migration after Salmonella treatment. Salmonella was found to influence the levels of phosphate-protein kinase B (P-AKT) and phosphate-extracellular regulated protein kinases (P-ERK), which are involved in heparanase expression. Salmonella reduced the heparanase expression induced upregulating PERK and PAKT signaling pathways. The mice bearing an experimental metastasis tumor model was used to evaluate the anti-tumor metastatic effects of Salmonella. Compared with the control group, Salmonella significantly reduced the number of metastatic nodules and enhanced survival. The results of our study indicate that Salmonella plays a vital role in the inhibition of tumor metastasis through the downregulation of heparanase.     

乙酰肝素酶siRNA对部分肝切除大鼠术后肝再生的抑制作用

目的探讨乙酰肝素酶(HPSE)siRNA对部分肝切除大鼠术后肝再生的抑制作用。方法设计合成HPSE siRNA,以vivo-jetPEI-Gal为载体转染70%肝切除大鼠,以逆转录-聚合酶链反应(RT-PCR)检测转染后HPSE mRNA表达,以Western blot检测AFP、ALB、CK-18和CK-19蛋白表达,以原位末端标记法(Tunel)检测肝细胞凋亡,以免疫组化检测肝组织增殖细胞核抗原(PCNA)表达,并测定肝再生率。结果与各对照组相比,siRNA干扰组HPSE mRNA表达明显降低,AFP、ALB、CK-18和CK-19蛋白表达明显降低,肝细胞凋亡指数明显增高,增殖指数明显降低,肝再生率明显降低,差别显著(P0.05)。结论 siRNA沉默HPSE表达后,部分肝切除大鼠术后肝再生受到明显抑制,提示HPSE对肝再生具有调控作用。     

小鼠局灶性脑缺血后乙酰肝素酶的表达

目的检测乙酰肝素酶（Hpa）在实验小鼠局灶性脑缺血后的表达,探讨其表达时间和表达位置变化的规律及其意义。方法用结扎大脑中动脉皮质支的方法制作小鼠局灶性脑缺血模型;碰用实时荧光定啦PCR方法检测缺血后Hpa、血管内皮生长因子（VEGF）和血管生成素2（Ang-2）基因的表达;应用免疫荧光双染（Hpa／CD105、Hpa／GFAP、Hpa／BrdU）的方法进一步观察Hpa蛋白表达的情况。结果实验组小鼠脑缺血后,与假手术组比较,HpamRNA于缺血后3d开始在缺血区的表达明显增高（P〈0．05）,14d达到相对高峰（与1、3、7d和假手术组比较,P〈0．05—0．01）。VEGFmRNA和Ang-2mRNA在脑缺血后3d的表达亦明鼹增高（与假手术组比较,P〈0．01）,其中VEGF的表达水平持续增高,而Ang-2则随后降低：免疫荧光双染显示,Hpa在脑缺血后7d时主要表达于新生的微血管,14d时主要与星形胶质细胞共表达：结论Hpa表达时间和表达位置的变化提示,Hpa参与了脑缺血后血管新生和组织修复的过程,推测Hpa可能通过促进VEGF的表达以及星形胶质细胞的增生而发挥其作用．     

宫颈上皮内瘤变与宫颈癌中乙酰肝素酶的表达

目的:探讨乙酰肝素酶在宫颈黏膜上皮内瘤变(CIN)与宫颈鳞癌中的表达.方法:应用免疫组化SP法检测乙酰肝素酶在56例CIN(CINⅠ12例,CINⅡ26例,CINⅢ18例),宫颈鳞癌54例(伴有淋巴结转移的20例)中的表达.结果:在CINⅠ,CINⅡ,CIN Ⅲ组织中乙酰肝素酶阳性表达率分别为33.33%,38.48%,44.44%,宫颈鳞癌中乙酰肝素酶阳性表达率为48.14%.CIN与宫颈鳞癌的表达对比统计学无意义(P＞0.05).20例宫颈鳞癌伴淋巴结转移者,其癌组织内乙酰肝素酶阳性表达(70.00%)明显高于无转移组(35.29%),差异有显著性(P＜0.01)结论:乙酰肝素酶表达与宫颈鳞癌发生、发展无关.与宫颈鳞癌淋巴结转移有关.     

肝素酶在非小细胞肺癌中的表达及其临床意义

背景与目的已有的研究表明,肝素酶的表达与肿瘤的发生有关。本研究通过肝素酶在非小细胞肺癌(NSCLC)中的表达情况,探讨肝素酶与NSCLC临床病理及其预后的关系。方法采用免疫组化检测66例NSCLC和15例相应癌旁正常肺组织中肝素酶的表达。结果66例NSCLC组织中肝素酶表达阳性率为74.2%,而正常支气管和肺泡上皮中无阳性表达;肝素酶表达与肺癌的临床分期(P=0.042)、淋巴结转移(P=0.005)有密切关系;肝素酶阳性表达组患者的5年生存率明显低于阴性组(P=0.025)。结论肝素酶在人肺癌组织中的表达较正常肺组织明显上调,并与肺癌的临床分期、淋巴结转移和患者的预后相关,但不能作为患者预后评估的独立指标。