E-选择素及其配体表达对肝癌转移的临床意义

目的 探讨E-选择索(E-selectin)及其配体在肝癌细胞与内皮细胞黏附中的作用和二者表达的临床意义;并筛选可能阻断黏附作用的药物. 方法 收集78例肝细胞癌患者的临床资料,采用免疫组织化学方法检测E-selectin、sLeX、sLeA、CD44v6的表达情况,按临床特征分组并比较各组E-selectin、sLeX、sLeA、CD44v6的阳性表达率差异.体外实验采用实时荧光定量PCR检测E-selectin和细胞间黏附分子(ICAM)-1在ED25、ECV304两种内皮细胞中的表达;黏附实验检测肝癌细胞HepG2与ED25、ECV304两种内皮细胞的黏附活性,并检测不同药物对黏附作用的影响.对临床资料数据采用χ2检验、Kaplan-Meier生存分析与Wilcoxon检验;对体外实验计量数据进行方差分析.结果 E-selectin在肝癌病灶内血管内皮细胞中的阳性表达率为70.51%,sLeX、sLeA、CDJ44v6在肝癌细胞中的阳性表达率分别为64.10%、69.23%、62.90%.sLeX、sLeA、CD44v6表达阴性患者的平均生存期显著长于阳性患者(P值均<0.05).有门静脉癌栓、术前肝外转移、卫星病灶、术后3个月内复发各组E-selectin、sLeX、sLeA的阳性表达率差异均有统计学意义(P值均<0.05);而肿瘤大小、血清AFP水平均与E-selectin、sLeX、sLeA的阳性表达率无相关性;病理分级Ⅲ～Ⅳ级组与Ⅰ～Ⅱ级组的E-selectin和CD44v6的阳性表达率差异均有统计学意义(P值均<0.05).CD44v6的表达特点是术后3个月内复发者有较高的阳性率,有卫星灶者阳性率也较高.E-selectin和ICAM-1可高表达于活化后的ED25和ECV304细胞;且能介导HepG2细胞与ED25和ECV304细胞的黏附,地塞米松,丹参酮ⅡA都具有较强的抗黏附能力,且抗黏附作用随着浓度的增加而增强.结论 E-selectin、sLeX.sLeA、CD44v6的表达与患者的预后,门静脉癌栓等临床特征密切相关,可作为临床预后和术后早期复发的评估依据.E-selectin、ICAM-1及其配体可增强肝癌细胞与内皮细胞的黏附,促进肝癌转移;地塞米松、丹参酮ⅡA可阻断其黏附作用.     

E-选择素及其配体表达对肝癌转移的临床意义

目的 探讨E-选择索(E-selectin)及其配体在肝癌细胞与内皮细胞黏附中的作用和二者表达的临床意义;并筛选可能阻断黏附作用的药物. 方法 收集78例肝细胞癌患者的临床资料,采用免疫组织化学方法检测E-selectin、sLeX、sLeA、CD44v6的表达情况,按临床特征分组并比较各组E-selectin、sLeX、sLeA、CD44v6的阳性表达率差异.体外实验采用实时荧光定量PCR检测E-selectin和细胞间黏附分子(ICAM)-1在ED25、ECV304两种内皮细胞中的表达;黏附实验检测肝癌细胞HepG2与ED25、ECV304两种内皮细胞的黏附活性,并检测不同药物对黏附作用的影响.对临床资料数据采用χ2检验、Kaplan-Meier生存分析与Wilcoxon检验;对体外实验计量数据进行方差分析.结果 E-selectin在肝癌病灶内血管内皮细胞中的阳性表达率为70.51%,sLeX、sLeA、CDJ44v6在肝癌细胞中的阳性表达率分别为64.10%、69.23%、62.90%.sLeX、sLeA、CD44v6表达阴性患者的平均生存期显著长于阳性患者(P值均<0.05).有门静脉癌栓、术前肝外转移、卫星病灶、术后3个月内复发各组E-selectin、sLeX、sLeA的阳性表达率差异均有统计学意义(P值均<0.05);而肿瘤大小、血清AFP水平均与E-selectin、sLeX、sLeA的阳性表达率无相关性;病理分级Ⅲ～Ⅳ级组与Ⅰ～Ⅱ级组的E-selectin和CD44v6的阳性表达率差异均有统计学意义(P值均<0.05).CD44v6的表达特点是术后3个月内复发者有较高的阳性率,有卫星灶者阳性率也较高.E-selectin和ICAM-1可高表达于活化后的ED25和ECV304细胞;且能介导HepG2细胞与ED25和ECV304细胞的黏附,地塞米松,丹参酮ⅡA都具有较强的抗黏附能力,且抗黏附作用随着浓度的增加而增强.结论 E-selectin、sLeX.sLeA、CD44v6的表达与患者的预后,门静脉癌栓等临床特征密切相关,可作为临床预后和术后早期复发的评估依据.E-selectin、ICAM-1及其配体可增强肝癌细胞与内皮细胞的黏附,促进肝癌转移;地塞米松、丹参酮ⅡA可阻断其黏附作用.     

心血康对血管紧张素Ⅱ诱导血管内皮细胞表达ICAM-1的影响

在体外培养的脐静脉内皮细胞株ECV304中,给予不同浓度的心血康作用2h后,再予血管紧张素Ⅱ（Angli）作用18h诱导内皮细胞表达细胞间黏附分子-1（ICAM-1）,采用流式细胞术检测ICAM-1的表达,用普通光学显微镜测淋巴细胞黏附率。结果显示,AngⅡ可上调内皮细胞表达ICAM-1,提高淋巴细胞黏附率;而心血康可抑制该表达,降低淋巴细胞黏附率,且呈浓度依赖性。     

膀胱移行细胞癌组织中CD44v6的表达及意义

采用免疫组化SABC法检测64例膀胱移行细胞癌和癌旁膀胱组织中的CD4v6。结果显示,64例膀胱移行细胞癌组织中CD44v6阳性表达率为56.3%,癌旁膀胱组织中未见CD44v6蛋白表达;CD44v6表达水平与膀胱移行细胞癌组织学分级、临床分期及复发有关。认为CD44v6可作为判断膀胱移行细胞癌的恶性程度和复发的参考依据。     

CO2气腹对结肠癌细胞表面粘附分子表达影响的研究

目的 探讨不同压强持续性CO2气腹对结肠癌细胞表面粘附分子表达的影响.方法 建立体外气腹模型,选用人结肠癌细胞株SW1116,分别在不同压强医用CO2气体下暴露1 h,使用流式细胞技术和实时荧光定量PCR检测粘附分子E-cadherin,ICAM-1,CD44,CD44v6,E-selectin的表达.结果 荧光定量PCR结果显示SW1116经6 mm Hg压强的持续CO2气体处理后,ICAM-1和CD44v6出现表达增高;9 mm Hg及12 mm Hg CO2气体处理后,E-cadherin,CD44和CD44v6表达增高;15 mm Hg CO2气体处理后CD44v6表达增高,与处理前表达水平的差异均有统计学意义（P＜0.05）,上述粘附分子的表达在处理后72 h内均会降至处理前水平（P＞O.05）,或低于处理前水平（P＜0.05）.E-cadherin、CD44v6和ICAM-1随着压强增高,其表达量逐渐降低.结论 不同压强CO2气腹能对肿瘤细胞表面的粘附分子表达产生一过性双向影响,随CO2气腹压强的增高,可抑制粘附分子的表达.     

喉癌组织中CD44v5、CD44v6蛋白的表达变化及意义

目的观察喉癌组织中细胞黏附分子CD44v5、CD44v6的表达变化,并探讨其临床意义。方法分别采用免疫组化SP法检测78例喉癌(观察组)及14例癌旁正常组织(对照组)中的CD44v5、CD44v6蛋白。结果观察组CD44v5、CD44v6蛋白阳性52、55例,对照组分别为4、0例;两组比较,P均<0.05。CD44v5、CD44v6蛋白表达与喉癌的TNM分期、组织分化程度及淋巴结转移有关(P均<0.05)。结论喉癌组织中CD44v5、CD44v6蛋白表达增加,两者可作为喉癌转移及预后评估的生物学指标。     

Calphostin C抑制酶修饰低密度脂蛋白诱导的内皮-单核细胞黏附

目的 探讨酶修饰的低密度脂蛋白(E-LDL)诱导内皮-单核细胞黏附的机制及Calphostin C的干预效果.方法 采用体外培养和直接计数法观察荷脂ECV304内皮细胞黏附能力的变化;PepTag○ RAssay法定性内皮细胞胞膜蛋白激酶C(PKC)活性状态;RT-PCR和Western印迹检测细胞间黏附分子-1(ICAM-1)和抑制性蛋白κBα(I-κBα)表达的变化.结果 E-LDL呈剂量依赖性增强内皮-单核细胞间的黏附,其活化内皮促进黏附的理想剂量为20～40 μg/ml浓度.另外,E-LDL还可显著活化内皮细胞胞膜PKC,下调I-κBα的表达而增强ICAM-1的表达.应用PKC特异性抑制剂Calphostin C干预后,荷载25 μg/ml E-LDL 8 h的ECV304内皮细胞ICAM-1表达下调而I-κBα则反相上调,黏附能力也在Calphostin C达到100 nmol/L浓度时显著下调,细胞状态明显好转;且200～400 nmol/L Calphostin C基本可以逆转E-LDL对内皮细胞的影响.结论 内皮细胞黏附能力的强效促进剂E-LDL可能是通过PKC/NF-κB/ICAM-1发挥作用的,针对PKC靶酶的特异性抑制剂Calphostin C可以有效地抑制由E-LDL引起的内皮活化黏附增强.     

VEGF和CD44v6过表达与乳腺癌浸润转移的关系

目的检测血管内皮生长因子(VEGF)和转移相关黏附分子44v6(CD44v6)在乳腺癌中的表达情况,探讨两者与淋巴结转移的关系。方法采用免疫组化SP法检测92例乳腺浸润性癌中VEGF、CD44v6的表达情况。结果 VEGF在正常乳腺组织和乳腺癌中的阳性表达率分别为6.7%(3/45)和90.2%(83/92),且VEGF在淋巴结转移组中的阳性表达率(53/55)明显高于无淋巴结转移组(P<0.05)。CD44v6在正常乳腺组织及乳腺癌中的阳性表达率分别为8.9%(4/45)和85.9%(79/92),而且CD44v6在淋巴结转移组中的阳性表达率(51/55)明显高于无淋巴结转移组(P<0.05)。VEGF和CD44v6表达呈正相关关系(r=0.497,P<0.05)。结论 VEGF、CD44v6在乳腺癌组织中高表达,且均与淋巴结转移有关(P<0.05),两者可能在乳腺癌的远处转移中起协同作用。     

c-erbB-2、CD44v6的表达与非小细胞肺癌淋巴转移的关系

目的探讨癌基因c-erbB-2和细胞黏附分子CD44v6与非小细胞肺癌(NSCLC)淋巴结转移的关系。方法应用免疫组织化学SABC法检测65例原发性NSCLC组织中c-erbB-2和CD44v6基因蛋白的表达水平。结果c-erbB-2在有无淋巴结转移组的阳性表达率分别为73.7%和48.1%,两组比较差异有显著性(P<0.05)。CD44v6在有无淋巴结转移组的阳性表达率分别为78.9%和55.6%,两组比较差异有显著性(P<0.05)。在有淋巴结转移的NSCLC中,c-erbB-2和CD44v6基因蛋白的表达呈显著相关关系。结论c-erbB-2阳性表达和CD44v6阳性表达可能均在NSCLC的淋巴结转移中起重要的促进作用,且二者的作用具有协同性。对c-erbB-2和CD44v6的检测有助于判断NSCLC的淋巴结转移。     

蚯蚓纤溶酶降低胃癌细胞与血管内皮细胞粘附性及CD44v6表达

目的研究蚯蚓纤溶酶对人胃癌细胞株MGC803的增殖抑制作用以及对胃癌细胞与血管内皮细胞粘附性和粘附分子CD44v6表达的影响。方法采用四甲基偶氮唑盐（MTT）法检测蚯蚓纤溶酶对人胃癌细胞株MGC803增殖的影响；体外粘附实验观察蚯蚓纤溶酶对胃癌细胞MGC803与血管内皮细胞ECV304粘附性的影响；流式细胞术观察蚯蚓纤溶酶作用于MGC803细胞后CD44v6蛋白表达的变化；RT—PCR法检测蚯蚓纤溶酶对MGC803细胞CD44v6mRNA表达的影响。结果蚯蚓纤溶酶各用药组（2、4、8uku／ml）对MGc803细胞均有抑制生长作用（P〈0．01），且在一定浓度范围内呈剂量依赖关系。蚯蚓纤溶酶4、8uku／ml作用于MGC803细胞48h后，与对照组相比，胃癌细胞与血管内皮细胞的粘附性明显下降（P〈0．01）。2、4、8uku／ml蚯蚓纤溶酶作用后，CD44v6蛋白表达水平呈剂量依赖性下降，平均荧光强度分别为84．80±2．14、30．69±3．66、21．75±2．25，与对照组相比，差异均有统计学意义（P〈0．05或0．01）。DNA半定量分析表明，各实验组CD44v6 mRNA表达也明显减少（P〈0．01）。结论蚯蚓纤溶酶有抑制胃癌细胞增殖的作用，并可降低胃癌细胞与血管内皮细胞的粘附性、降低胃癌细胞粘附分子CD44v6的表达，提示蚯蚓纤溶酶可能具有潜在抗肿瘤转移作用。     

细胞粘附分子在胆囊癌中表达的意义

研究胆囊癌组织中细胞粘附分子CEA和CD44v6蛋白表达的意义。用免疫组化法检测 44例胆囊癌组织中CEA和CD44v6蛋白的表达。 44例胆囊组织中CEA和CD44v6蛋白表达的阳性率分别为 75 %和 48% ;两者的表达与肿瘤浸润外膜和淋巴结转移均呈正相关 (P <0 0 5 ) ;胆囊癌组织中CEA和CD44v6蛋白呈协同表达 (P<0 0 5 )。CEA和CD44v6蛋白与胆囊癌的发展和淋巴结转移密切相关。     

Fluid shear stress modulates surface expression of adhesion molecules by endothelial cells

We investigated the effect of hemodynamic shear forces on the expression of adhesive molecules, E-selectin, and intercellular adhesion molecule-1 (ICAM-1) on human umbilical vein endothelial cells (HUVEC) exposed to laminar (8 dynes/cm2) or turbulent shear stress (8.6 dynes/cm2 average), or to a static condition. Laminar flow induced a significant time-dependent increase in the surface expression of ICAM- 1, as documented by flow cytometry studies. Endothelial cell surface expression of ICAM-1 in supernatants of HUVEC exposed to laminar flow was not modified, excluding the possibility that HUVEC exposed to laminar flow synthetize factors that upregulate ICAM-1. The effect of laminar flow was specific for ICAM-1, while E-selectin expression was not modulated by the flow condition. Turbulent flow did not affect surface expression of either E-selectin or ICAM-1. To evaluate the functional significance of the laminar-flow-induced increase in ICAM-1 expression, we studied the dynamic interaction of total leukocyte suspension with HUVEC exposed to laminar flow (8 dynes/cm2 for 6 hours) in a parallel-plate flow chamber or to static condition. Leukocyte adhesion to HUVEC pre-exposed to flow was significantly enhanced, compared with HUVEC maintained in static condition (233 +/- 67 v 43 +/- 16 leukocytes/mm2, respectively), and comparable with that of interleukin-1 beta treated HUVEC. Mouse monoclonal antibody anti-ICAM-1 completely blocked flow-induced upregulation of leukocyte adhesion. Interleukin-1 beta, which upregulated E-selectin expression, caused leukocyte rolling on HUVEC that was significantly lower on flow- conditioned HUVEC and almost absent on untreated static endothelial cells. Thus, laminar flow directly and selectively upregulates ICAM-1 expression on the surface of endothelial cells and promotes leukocyte adhesion. These data are relevant to the current understanding of basic mechanisms that govern local inflammatory reactions and tissue injury.     

Binding of gastrointestinal tumor cells to endothelial E- and P-selectin adhesion receptors leads to transient down-regulation of sLeX ligands in vitro

BACKGROUND AND AIMS. The prognostic relevance of sialyl Lewis X (sLeX) expression in colorectal and gastric cancer and its relevance to the hematogenous phase of tumor invasion is controversial. This study was designed to evaluate sLeX expression during tumor cell-endothelial cell interaction in vitro. METHODS. Adhesion and transendothelial penetration of MKN45, PaCa-2, WiDr, or Dan-G cells was analyzed by combined phase contrast-reflection interference contrast microscopy. In parallel, kinetics of membranous sLeX expression were examined fluorimetrically. To identify factor(s) which may be responsible for sLeX expression during tumor invasion tumor cells were treated with soluble immunomodulators, isolated endothelial plasma membranes, or E-selectin or P-selectin IgG fusion proteins. sLeX was then analyzed by flow cytometry. RESULTS. Fluorometric quantification of sLeX demonstrated an inverse correlation between basal sLeX expression level and adhesion capacity of the tumor cells. Unexpectedly, sLeX was strongly down-regulated on tumor cell membranes in the course of heterophilic cell-cell contacts. The process occurred transiently, with a maximum effect 30-60 min after introducing tumor cells to the endothelial monolayer. Binding of tumor cells to immobilized E- and P-selectin IgG globulin chimeras was shown to be responsible for this phenomenon. CONCLUSION. A transient loss of sLeX is necessary for gastrointestinal tumor cells to invade endothelial cells. Due to the transient nature of the decrease in sLeX the controversy about the prognostic relevance of sLeX expression in colorectal and gastric cancer may be rooted in the stage of tumor invasion at the time of sLeX measurement.     

Higher CO2-insufflation pressure inhibits the expression of adhesion molecules and the invasion potential of colon cancer cells

AIM： To investigate the influence of CO2-insufflation pressure on adhesion, invasion and metastatic potential of colon cancer cells based on adhesion molecules expression. METHODS： With an/n vitro artificial pneumoperitoneum model, SW1116 human colon carcinoma cells were exposed to CO2-insufflation in 5 different pressure groups： 6 mmHg, 9 mmHg, 12 mmHg, 15 mmHg and control group, respectively for 1 h. Expression of E-cadherin, ICAM-I, CD44 and E-selectin was meas- ured at 0, 12, 24, 48 and 72 h after CO2-insufflation using flow cytometry. The adhesion and invasion capacity of SW1116 cells before and after exposure to CO2-insufflation was detected by cell adhesion/invasion assay in vitro. Each group of cells was injected intraperitoneally into 16 BALB/C mice. The number of visible abdominal cavity tumor nodules, visceral metas-tases and survival of the mice were recorded in each group. RESULTS： The expression of E-cadherin, ICAM-1, CD44 and E-selectin in SWl116 cells were changed significantly following exposure to CO2 insufflation at different pressures （P 〈 0.05）. The expression of E-cadherin, CD44 and ICAM-1 decreased with increasing CO2-insufflation pressure. The adhesive/ invasive cells also decreased gradually with increasing pressure as determined by the adhesion/invasion assay. In animal experiments, the number of abdominal cavity tumor nodules in the 15 mmHg group was also significantly lower than that in the 6 mmHg group （29.7± 9.91 vs 41.7±14.90, P = 0.046）. However, the survival in each group was not statistically different. CONCLUSION： CO2-insufflation induced a temporary change in the adhesion and invasion capacity of cancer cells in vitro. Higher CO2-insufflation pressure inhibited adhesion, invasion and metastatic potential in vitro and in vivo, which was associated with reduced expression of adhesion molecules.     

胆囊癌组织中CD44v6和nm23的表达及其意义

目的研究胆囊癌组织中CD44v6、nm23蛋白表达及意义和两者之间的关系。方法用免疫组化的方法检测 36例胆囊癌和27例癌旁上皮组织中CD44v6和nm23蛋白的表达。结果胆囊癌组织中CD44v6和nm23的阳性率分别为41.7％和72.2％,均与癌旁上皮的阳性率有显著性差异:CD44v6的表达与胆囊癌的淋巴结转移呈正相关,而与组织学类型、临床分期无关;nm23的表达与胆囊癌的淋巴结转移、临床分期呈负相关,而与组织学类型无关。CD44v6和nm23蛋白的表达呈负相关。结论 CD44v6和nm23蛋白可能在胆囊癌的形成、淋巴结转移和病程进展等方面起着重要作用,且两者呈互相拮抗的关系,可作为判断胆囊癌侵袭和淋巴结转移的标记物。     

胃癌中CD15s抗原和CD44v6基因蛋白的表达及其意义

[目的 ]探讨唾液酸化的路易斯寡糖 X(CD15s)抗原和CD4 4v6基因蛋白表达与胃癌临床病理指标和患者预后的关系。 [方法 ]应用高敏感性的催化信号放大免疫组化技术 ,对早期胃癌 17例、中期胃癌 2 1例和晚期胃癌5 7例组织进行CD15s抗原和CD4 4v6蛋白检测 ,并结合肿瘤的病理学指标和临床随访资料进行分析。 [结果 ]在胃癌中 ,CD15s和CD4 4v6的表达阳性率分别为 86 .3%和 82 .1%。且晚期胃癌明显高于早、中期胃癌 (P <0 .0 5 )。CD15s和CD4 4v6表达水平具有一致性 ,均与胃癌浆膜浸润、淋巴结转移和患者预后呈正相关 (P <0 .0 5 )。 [结论 ]CD15s和CD4 4v6表达与胃癌转移和患者生存期密切相关 ,检测CD15s和CD4 4v6的表达可作为判断胃癌预后的参考指标。     

尼美舒利抑制结肠癌侵袭和转移的作用机制研究

目的观察选择性环氧合酶-2(COX-2)抑制剂尼美舒利对结肠癌细胞Caco-2同质性黏附能力,E-钙黏蛋白和CD44v6表达的影响.探讨其在抑制肿瘤侵袭和转移中的作用机制.方法培养结肠癌细胞细胞株Caco-2,用不同浓度的尼美舒利进行干预,观察细胞同质性黏附能力的变化.用Western blot检测E-钙黏蛋白表达的变化,用免疫细胞化学法检测CD44v6的变化.结果尼美舒利能增加Caco-2细胞的同质性黏附能力,增加E-钙黏蛋的表达,并呈剂量依赖性.结论尼美舒利可以通过增强结肠癌细胞的同质性黏附能力,增加E-钙黏蛋白的表达,减少CD44v6的表达来抑制肿瘤细胞的侵袭和转移.