乳腺癌组织中HER-2、VEGF和Ki67的表达及临床意义

目的探讨人表皮生长因子受体-2(HER-2)、血管内皮生长因子(VEGF)和Ki67在乳腺癌组织中的表达及其与临床病理因素的关系。方法采用免疫组化法检测49例乳腺癌患者组织中HER-2、VEGF和Ki67的表达。结果 HER-2、VEGF和Ki67在乳腺癌组织中的阳性表达率分别为36.7%、69.4%、79.6%;HER-2在乳腺癌组织学分级和淋巴结转移中的表达差异有统计学意义(P<0.05);VEGF在乳腺癌淋巴结转移中的表达有显著差异(P<0.05);Ki67表达在乳腺癌组织学分级中的差异有统计学意义(P<0.05)。结论 HER-2、VEGF和Ki67与乳腺癌的发生、发展密切相关,联合检测可作为判定乳腺癌预后的评估指标。

携抗VEGFR2抗体PLGA靶向超声造影剂的制备及体外寻靶实验

目的制备携抗血管内皮细胞生长因子受体2(VEGFR2)抗体聚乳酸羟基乙酸(PLGA)靶向超声造影剂,并考察其体外寻靶能力与超声显像性能。方法通过改进的双乳化溶剂挥发法制备高分子材料PLGA纳米粒子,利用扫描电子显微镜对其一般特性进行表征,并进一步用碳二亚胺法将造影剂与抗VEGFR2抗体耦联制备靶向超声造影剂,使用激光共聚焦扫描显微镜对其体外寻靶能力进行初步评估,使用高频超声诊断仪观察体外显像效果。结果 PLGA超声造影剂粒子呈规则球形、大小均一、分散性好;在体外寻靶实验中,携抗VEGFR2抗体PLGA靶向造影剂能够较多并牢固的聚集到血管肉瘤内皮细胞(SVR)表面;体外超声成像实验中,携抗VEGFR2抗体PLGA靶向超声造影剂呈点状细密高回声,后方回声无衰减。结论本研究成功制备携抗VEGFR2抗体PLGA靶向超声造影剂,能够在体外与VEGFR2高表达的血管肉瘤内皮细胞特异性靶向结合,且体外超声显像效果良好。

Protective effects of a novel drug RC28-E blocking both VEGF and FGF2 on early diabetic rat retina

AIM： To investigate protective effects of a novel recombinant decoy receptor drug RC28-E on retinal damage in early diabetic rats. METHODS： The streptozotocin （STZ）-induced diabetic rats were randomly divided into 6 groups： diabetes mellitus （DM） group （saline, 3 μL/eye）; RC28-E at low （0.33 μg/μL, 3 μL）, medium （1 μg/μL, 3 μL）, and high （3 μg/μL, 3 μL） dose groups; vascular endothelial growth factor （VEGF） Trap group （1 μg/μL, 3 μL）; fibroblast growth factor （FGF） Trap group （1 μg/μL, 3 μL）. Normal control group was included. At week 1 and 4 following diabetic induction, the rats were intravitreally injected with the corresponding solutions. At week 6 following the induction, apoptosis in retinal vessels was detected by TUNEL staining. Glial fibrillary acidic protein （GFAP） expression was examined by immunofluorescence. Blood-retinal barrier （BRB） breakdown was assessed by Evans blue assay. Ultrastructural changes in choroidal and retinal vessels were analyzed by transmission electron microscopy （TEM）. Content of VEGF and FGF proteins in retina was measured by enzyme linked immunosorbent assay （ELISA）. The retinal expression of intercellular cell adhesion molecule-1 （ICAM-1）, tumor necrosis factor-α （TNF-α）, VEGF and FGF genes was examined by quantitative polymerase chain reaction （qPCR）. RESULTS： TUNEL staining showed that the aberrantly increased apoptotic cells death in diabetic retinal vascular network was significantly reduced by treatments of medium and high dose RC28-E, VEGF Trap, and FGF Trap （all P〈0.05）, the effects of medium and high dose RC28-E or FGF Trap were greater than VEGF Trap （P〈0.01）. GFAP staining suggested that reactive gliosis was substantially inhibited in all RC28-E and VEGF Trap groups, but the inhibition in FGF Trap group was not as prominent. Evans blue assay demonstrated that only high dose RC28-E could significantly reduce vascular leakage in early diabetic retina （P〈0.01）. TEM revealed that the ultrastructures in choroidal and retinal vessels were damaged in early diabetic retina, which was ameliorated to differential extents by each drug. The expression of VEGF and FGF2 proteins was significantly upregulated in early diabetic retina, and normalized by RC28-E at all dosages and by the corresponding Traps. The upregulation of ICAM-1 and TNF-α in diabetic retina was substantially suppressed by RC28-E and positive control drugs. CONCLUSION： Dual blockade of VEGF and FGF2 by RC28-E generates remarkable protective effects, including anti-apoptosis, anti-gliosis, anti-leakage, and improving ultrastructures and proinflammatory microenvironment, in early diabetic retina, thereby supporting further development of RC28-E into a novel and effective drug to diabetic retinopathy （DR）.

藤黄酸对人结肠癌SW480细胞裸鼠移植瘤生长及血管生成的影响

目的探讨藤黄酸对人结肠癌SW480细胞裸鼠移植瘤生长和肿瘤血管生成的抑制作用及藤黄酸对血管内皮生长因子受体2(VEGFR2)表达的影响,初步探讨藤黄酸抗结直肠癌的作用机制。方法建立人结肠癌SW480细胞裸鼠皮下移植瘤模型,15 d后开始用不同浓度藤黄酸治疗,28 d后处死,测定裸鼠移植瘤的体积和质量,计算其抑瘤率;HE染色观察肿瘤坏死、微血管形态的变化;提取组织中总RNA和蛋白,RT-PCR检测肿瘤组织中VEGFR2 mRNA水平,免疫组织化学法检测肿瘤组织中VEGFR2蛋白表达水平和肿瘤微血管密度(MVD)。结果 (1)对照组,藤黄酸低、中、高剂量组移植瘤体积分别为(691±67.2)、(481±62.9)、(377±37.0)、(190±67.9)mm3,瘤质量分别为(0.681±0.072)、(0.473±0.062)、(0.377±0.044)、(0.192±0.068)g,藤黄酸低、中、高剂量组瘤体积、瘤质量均低于对照组(均P<0.05);藤黄酸低、中、高剂量组抑瘤率分别为31%、45%、72%;(2)HE染色可见藤黄酸组肿瘤组织出现凝固性坏死,肿瘤新生血管闭锁;(3)与对照组相比,藤黄酸组VEGFR2 mRNA表达丰度和蛋白水平明显降低,并呈显著量效关系;(4)对照组MVD明显高于藤黄酸组(P<0.05)。结论藤黄酸能够抑制人结肠癌SW480细胞裸鼠移植瘤的生长,能抑制结肠癌移植瘤内新生血管内皮细胞的生长,其机制可能与抑制VEGFR2的表达有关。

通过阻断花生四烯酸代谢途径抑制胰腺癌细胞增殖

目的:探讨通过阻断花生四烯酸(arachidonic acid,AA)代谢途径抑制胰腺癌细胞增殖.方法:将胰腺癌细胞SW1990分为对照组,M K886干预组、塞莱昔布(C e l e c o x i b)干预组,MK886+Celecoxib干预组,用RT-PCR法检测细胞白三烯B4受体1(leukotriene B4receptor 1,BLT1)mRNA,血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA的表达量变化,用Western blot检测磷酸化-Erk(phosphorylated-extracellular regulated protein,p-Erk)表达量变化.结果:MK886作用下,BLT1 mRNA、VEGF mRNA等表达量均减少(P<0.01),p-Erk表达量明显减少(P<0.05),Celecoxib作用下,VEGF mRNA表达量明显减少(P<0.01),BLT1 mRNA表达与对照组无明显差异,p-Erk表达量与MK886组比较明显增加(P<0.01),MK886+80?mol/L Celecoxib作用下,BLT1 mRNA、VEGF mRNA表达量明显减少(P<0.01),p-Erk表达量与对照组无明显差异.结论:花生四烯酸的两条代谢途径均与胰腺癌的发生及增殖均有密切关系,而抑制5-脂氧合酶(5-lipoxygenase)途径较环氧化酶2(cyclooxygenase 2)途径相比,抑制肿瘤细胞增殖作用更强.

血管内皮生长因子受体-1和血管内皮生长因子受体-2在子癎前期中的表达及临床意义

目的观察胎盘组织中血管内皮生长因子受体-1（VEGFR-1）、VEGFR-2、血浆中可溶性血管内皮生长因子受体-1（sVEGFR-1）及sVEGFR-2的表达水平，并探讨其在子癎前期的临床检测意义。方法ELISA检测正常妊娠期妇女和子癎前期患者血浆中的sVEGFR-1和sVEGFR-2的表达；RT-PCR检测正常妊娠期妇女和子癎前期患者胎盘中VEGFR-1、VEGFR-2、sVEGFR-1和sVEGFR-2mRNA的表达。结果ELISA检测结果显示，正常妊娠组分娩后较分娩前血浆中sVEGFR-1[（4．55±0．31）ng／mlvs（12．33±1．52）ng／m1]水平降低，子癎前期组分娩后血浆中sVEGFR．1[（8．13±0．74）ng／mlvs（77．25±9．47）ng／m1]水平亦降低，两组分娩前后数据变化比较差异有统计学意义（P〈0．01）；正常妊娠组分娩前后血浆中sVEGFR-2分别为（8．74±1．24）ng／ml与（6．43±0．55）ng／ml，而子痫前期组分别为（5．69±0．75）ng／ml与（4．96±0．67）ng／ml；RT-PCR检测结果与其-致。结论子癎前期患者分娩后VEGFR-1血浆水平迅速下降，而VEGFR-2持续低水平表达，提示VEGFR-1可能与子癎前期发病率有关，而子癎前期患者血浆中VEGFR-2水平的降低可作为内皮功能障碍的标志。

血管内皮生长因子及其受体各亚型在心肌梗死大鼠体内的差异性表达

目的研究血管内皮生长因子及其受体（VEGFR）各亚型在心肌梗死（MI）后的心肌修复和重构过程中的表达。方法构建大鼠心肌梗死模型，采用实时荧光定量PCR（RT—PCR）和West-emBlot法测定建模前后VEGF各亚型的表达。结果VEGF的所有亚型在正常大鼠心肌细胞中都会表达；与假手术对照大鼠相比，在心肌梗死大鼠心肌内，VEGF-A在MIId时表达为对照组的0．89±0．04倍，42d后显著降低，最低为0．25±0．03倍；VEGF—B在MI中受到显著抑制，最低为对照组的0．09±0．04倍；而VEGF-C和VEGF—D的表达则出现了显著的增加，分别为对照组的5．31±0．21倍和9．24±0．47倍。在MI大鼠心脏内，VEGFR-1和VEGFR~的表达显著减少，分别为对照组的0．11±0．02倍和0．14±0．04倍。而VEGFR-3，主要在血管中的水平则显著增加，是对照组的6．81±0．42倍。结论VEGF和VEGFR的各亚型在心肌梗死大鼠心脏内的表达发生显著性变化，VEGF—A可能是启动血管再生反应的关键调节因子，VEGF—D也是参与血管再生的重要因子，而VEGF—B并不是心肌修复和重构的关键因子。