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。     

Heparanase expression increases throughout the endometrial hyperplasia-cancer sequence.

OBJECTIVE: To assess the expression of heparanase in the different stages leading to endometrial cancer. METHODS: The 38 examined specimens included adenocarcinoma, hyperplasia, and normal endometrium specimens. Heparanase, estrogen, and progesterone receptor expressions were analyzed immunohistochemically and the intensity was scored. RESULTS: Secretory normal endometrium and simple hyperplasia specimens expressed the lowest mean values of expression (1.00 and 0.63, respectively); the complex hyperplasia specimens and G2 endometrioid adenocarcinoma showed the highest values of expression (2.33 and 2.71, respectively). A linear trend (P=0.005) of heparanase expression was observed when comparing the normal endometrium and simple hyperplasia group with the complex hyperplasia+G1 carcinoma group and the G2+G3 carcinoma group. Evaluation of atrophic and inactive endometrium compared with papillary serous carcinomas yielded no significant differences. We found no significant correlation between heparanase expression and estrogen receptor or progesterone receptor expression. CONCLUSION: Heparanase expression was tightly regulated in endometrial tumorigenesis.     

阻滞真核细胞起始因子-4E对膀胱癌细胞乙酰肝素酶表达的影响

目的 探讨膀胱癌细胞真核细胞起始因子-4E( eIF-4E)与乙酰肝素酶表达的关系.方法 脂质体包裹eIF-4E于mRNA翻译起始点互补的反义寡核苷酸(asODN)及相应对照的正义寡核苷酸(sODN)以100、200 pmol/L两种浓度分别转染膀胱癌EJ细胞,逆转录-聚合酶链反应(RT-PCR)检测eIF-4E转录水平的改变.荧光定量PCR和Western blot分别检测乙酰肝素酶mRNA及蛋白表达.结果 asODN转染显著抑制eIF-4E基因水平.eIF-4E被阻滞后,乙酰肝素酶mRNA水平及蛋白表达量:asODN1组(0.276±0.050、0.255±0.028)及asODN2组(0.195±0.022、0.160±0.032),均显著低于空白对照组(0.896±0.057、0.603±0.029)及sODN1组(0.867±0.099、0.572±0.029)和sODN2组(0.879±0.087、0.560±0.035,P＜0.01).结论 asODN转染膀胱癌EJ细胞可以阻滞eIF-4E基因表达,阻滞eIF-4E后,膀胱癌乙酰肝素酶mRNA水平及蛋白表达均明显降低.     

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.     

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.     

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.     

硼替佐米对多发性骨髓瘤细胞乙酰肝素酶表达及迁移能力的影响

目的: 研究硼替佐米对多发性骨髓瘤细胞株U266的乙酰肝素酶(HPA)表达及迁移能力的影响,并探讨其作用机制。方法: 体外培养U266细胞,以不同浓度的硼替佐米进行处理,采用CCK-8检测细胞活力,用RT-PCR方法检测HPA mRNA水平的变化,用Western blotting方法检测HPA蛋白及IκB表达水平的变化,以Transwell方法检测细胞迁移能力的变化。结果: U266细胞表达HPA,以0、3.125、12.5及50 nmol/L浓度的硼替佐米处理U266细胞48 h,HPA mRNA及蛋白的表达逐渐减少(P<0.01),而细胞迁移能力逐渐减弱(P<0.01)。同时,HPA的蛋白表达水平与IκB的表达水平呈负相关(P<0.01)。结论: 硼替佐米通过抑制HPA的表达抑制骨髓瘤细胞的迁移能力,其机制可能是通过NF-κB信号途径进行调控的。     

乙酰肝素酶反义寡核苷酸对人乳腺癌细胞株侵袭力的抑制作用

目的　探讨乙酰肝素酶 (HPSE)反义寡脱氧核苷酸 (AS ODN )对人乳腺癌细胞株侵袭力的抑制活性。方法　设计合成分别互补于HPSEmRNA 5 '未翻译区和起始密码区的AS ODN1、AS ODN2及相应对照NS ODN1、NS ODN 2 ,在脂质体介导下以 2 5 0nmol/L终浓度转染人乳腺癌MDA43 5细胞 ,以RT PCR和Westernblot分别检测HPSEmRNA和蛋白表达 ,以基质凝胶侵袭实验检测细胞侵袭力。结果　AS ODN 2组的HPSEmRNA相对表达量、蛋白表达量和侵袭细胞数(0 .62± 0 .0 4、12 .70± 1.70、61.0 0± 9.0 0 )与脂质体对照组 (1.60± 0 .0 4、3 6.2 0± 2 .10、178.0 0±17.0 0 )和NS ODN2组 (1.48± 0 .0 3、3 5 .5 0± 2 .3 0、174.0 0± 2 0 .0 0 )相比较均显著降低 (P <0 .0 1) ;而AS ODN 1组与脂质体对照组和NS ODN1组相比较差异均无显著性 (P >0 .0 5 )。结论　互补于起始密码区的HPSEAS ODN可通过下调HPSE表达显著抑制人乳腺癌细胞株MDA43 5的侵袭力。