血管内皮细胞特异性受体在肿瘤血管靶向治疗中的应用

目前已发现的内皮细胞特异性酪氨酸激酶受体家族主要有两种:一种是VEGF受体家族,包括Flt-1(VEGFR-1),Flk-1/KDR(VEGFR-2)和Flt-4(VEGFR-3);另一种是Tie家族,包括Tie-1和Tie-2(Tek)。它们都具有信号传导所必需的酪氨酸激酶活性,在生理及病理性血管再生中发挥着关键性作用。近来以VEGF受体及Tie受体作为靶点的肿瘤血管靶向治疗报道日渐增多,现综述如下: 1 VEGF受体血管内皮细胞生长因子(VEGF)是一种内皮细胞特异性的丝裂源,能在体内诱导血管生成,是作用最强的血管形成因子之一,它的高亲和力结合位点仅位于血管内皮细胞上的3种VEGF受体,即Flt-1,Flk-1/KDR和Flt-4,这3种受体主要分     

甲羟孕酮对子宫内膜电切术后EGFR、VEGF及KDR表达的影响

接受子宫内膜切除术(TCRE)的子宫内膜性疾病患者102例,随机分为A、B两组:A组47例,术后给予甲羟孕酮100 mg,2次/d,连续3个月;B组55例,术后不给甲羟孕酮;另取30例正常女性为对照组.术前及术后6个月用免疫组化法测定子宫内膜表皮生长因子受体(EGFR)与血管内皮生长因子(VEGF)及其受体(KDR)的表达情况.结果:所有患者子宫内膜EGFR、VEGF及其KDR的表达术前与术后相比,P<0.05,与对照组相比,P<0.05;术后6个月,所有患者子宫内膜的EGFR、VEGF及KDR的表达与正常对照组相比无显著性差异,但A组与B组相比,P<0.05.认为TCRE患者术后服用甲羟孕酮可调节子宫内膜EGFR、VEGF及KDR的表达,有助于抑制内膜性疾病的复发.     

急性白血病细胞bcl-2、c-myc和p53基因表达的研究

目的研究bcl-2、c-myc、p53基因在急性白血病(AL)细胞中的表达及其与AL的分型、临床特征、疗效及预后因素的关系.方法采用免疫组化方法检测54例初治AL骨髓细胞bcl-2、c-myc、p53蛋白表达情况,分析其与FAB分型、临床特征、疗效及预后因素的关系.结果AL骨髓细胞中bcl-2、c-myc及p53基因的表达显著高于正常对照组(P<0.05).bcl-2、c-myc、p53基因表达在急性髓系白血病(AML)与急性淋巴细胞白血病(ALL)差异不显著;在AML亚型中,bcl-2在M4及M5中的表达高于M1、M2和M3(P<0.05);而c-myc及p53的表达在各亚型中差异无显著性意义(P>0.05).bcl-2的表达与初诊时的白细胞数呈正相关(r=0.44,P<0.05),与疗效呈负相关(r=-0.40,P<0.05),与 c-myc的表达呈正相关(r=0.51,P<0.05);c-myc 的表达与初诊时的白细胞数呈正相关(r=0.50,P<0.05).结论bcl-2、c-myc、p53基因的紊乱可能在AL的发病中起着重要作用,还与白血病的某些临床特征、疗效及预后因素密切相关.     

NM-3含药血清对体外胃癌细胞株SGC-7901生长和VEGF及其受体KDR、Flt-1表达的影响

目的:探讨NM-3含药血清对胃癌SGC-7901细胞的抑制作用.方法:将SGC-7901胃癌细胞分成空白对照组、血清对照组、NM-3含药血清低、中、高剂量实验组共5组.在不同时间段以流式细胞仪检测细胞的凋亡率,分析细胞周期分布;应用逆转录聚合酶链式反应(RT-PCR)法检测各组胃癌细胞中VEGF及其受体KDR,Flt-1 mRNA的表达.结果:NM-3含药血清3组随着时间的延长凋亡率增高,呈时间剂量依赖性,其高、中剂量组在6h(16.80%±0.40%,25.20%±0.86%)、24h(26.95%±3.25%,43.62%±3.53%)、72 h(39.85%±4.10%,54.35%±5.42%)内的细胞早期凋亡率与空白,血清两个对照组相比有显著性差异(5_34%±0.28%,5.66%±0.72%:6.91%±1.06%,8.90%±0.86%;6.87%±1.24%,8.06%±0.78%;P<0.01).随着NM-3药物剂量的增加,G_0/G_1期细胞增多,而S期及G_2/M期细胞则相应减少.细胞培养72h后,NM-3含药血清高,中,低剂量组与两对照组比较可显著抑制VEGF及其受体KDR,Flt-1 mRNA的表达(P<0.05).在72h后细胞凋亡率,细胞周期以及胃癌细胞中VEGF及其受体KDR,Flt-1 mRNA的表达上,高剂量组与中低剂量组均有明显差异(P<0.05).结论:NM-3含药血清可抑制胃癌组织的血管生长,并可诱导胃癌细胞凋亡.     

胰岛素对牛胸主动脉内皮细胞血管内皮生长因子受体表达的影响及其与一氧化氮合酶活性的关系

目的　评价胰岛素对培养的牛胸主动脉内皮细胞血管内皮生长因子 (VEGF)受体flt 1、flk 1 KDR表达的影响及其与一氧化氮合酶 (NOS)的关系。方法　取新生的小牛胸主动脉 ,作血管内皮细胞原代及传代培养 ,取 4 6代培养细胞用于实验 ;应用不同浓度 ( 30mU L、30 0mU L、30 0 0mU L)的胰岛素和NOS抑制剂 (L NAME)干预培养过程 ,4 8h后取培养细胞 ,应用免疫组化法测定flt 1、flk 1 KDR和NOS3的表达水平。结果　不同浓度胰岛素孵育组后内皮细胞flt 1、flk 1 KDR的表达水平差异无显著性 ,L NAME孵育后各组内皮细胞flt 1、flk 1 KDR的表达水平较单纯胰岛素孵育组差异无显著性。结论　胰岛素对内皮细胞flt 1、flk 1 KDR的表达无直接影响 ;内皮细胞NOS3的活性不是内皮细胞VEGF受体flt 1、flk 1 KDR表达的主要影响因素     

血管内皮生长因子及其受体在肺气肿患者肺组织中的表达

目的探讨血管内皮生长因子(VEGF)及其受体2(VEGF受体2/KDR)在肺气肿患者肺组织中的表达及其与肺气肿的相关性。方法取35例行肺叶切除术患者[A组(吸烟伴肺气肿组)16例,B组(不吸烟肺功能正常组)14例,C组(吸烟但肺功能正常组)5例]的外周肺组织标本,ELISA法检测肺组织匀浆中VEGF的含量,免疫组化法检测KDR蛋白表达,RT PCR检测VEGF和KDRmRNA水平,TUNEL法检测肺泡隔细胞的凋亡。结果A组患者肺组织VEGF、KDR表达均低于B组(P<0.01),肺泡隔细胞凋亡率高于B组(P<0.01)。C组与B组相比,VEGF及KDR表达差异无统计学意义(P>0.05)。结论VEGF及KDR水平减少与肺泡隔细胞凋亡的增加可能与肺气肿的发生相关。     

急性淋巴细胞白血病 T淋巴细胞核仁组成区相关蛋白的检测及临床意义

目的探讨T淋巴细胞核仁组成区嗜银蛋白(Ag-NORS)在急性淋巴细胞白血病( ALL)的诊断及病情监测方面的临床意义.方法采用KL型肿瘤免疫图像分析系统及细胞培养液,以银染色试剂及其方法对正常人及ALL患者不同临床时期外周血T淋巴细胞核进行银染色和分析.结果正常人T淋巴细胞Ag-NORS含量明显高于ALL患者(P<0.05),正常人与ALL患者中完全缓解(CR)者和未完全缓解(NR)者比较,差异均有显著性意义(分别为P<0.05和P<0.01).结论T淋巴细胞Ag-NORS检测在ALL诊断及病情监测方面具有重要意义.     

VEGF及其受体KDR在结直肠腺瘤组织的表达水平及其与预后的关系

目的探讨VEGF及其受体KDR在结直肠腺瘤组织的表达水平及与预后的关系。方法采用免疫化学法对55例结直肠腺瘤病人体内VEGF及其受体KDR的表达水平进行检测。并随访5年生存率。结果两者在结直肠腺瘤的阳性表达率分别为68.6%,69.5%,且其表达率与年龄、患者无显著相关性(P>0.05),与肿瘤类型、是否有淋巴结转移关系密切(P<0.05)。VEGF及其受体KDR阳性者中5年生存率为45.5%、52.3%,阴性者5年内生存率为88.5%、87.9%,两组具有显著性差异(P<0.05)。Cox回归分析显示,对于结直肠腺瘤的患者,VEGF、KDR是患者死亡的危险因素,联合使用VEGF、KDR是预测结直肠腺瘤预后的良好指标。结论 VEGF及其受体KDR在结直肠腺瘤,发生发展中起到重要的作用,二者均阳性表达时提示患者预后差,二者联合可以作为评价结直肠腺瘤患者预后的有效指标。     

NF-kappa B在动脉粥样硬化中的作用

目的研究体内基因多显性转录核因子(NF-kappa B)在动脉粥样硬化疾病中病变的动脉内皮细胞中表达的情况及意义. 方法采用免疫组织化学染色S-P法检测30例动脉粥样硬化病变动脉血管和30例正常成年人动脉血管内皮细胞的NF-kappa BP65蛋白,细胞间粘附分子-1(ICAM-1)和血管内皮细胞粘附分子-1(VCAM-1)的表达水平. 结果 (1)与正常动脉血管内皮细胞相比较, 动脉粥样硬化病变动脉血管内皮细胞NF-kappa BP65蛋白,ICAM-1以及VCAM-1表达水平均呈显著性增强(P<0.001).(2)60例动脉血管内皮细胞检测数据提示NF-kappa B蛋白表达和ICAM-1表达呈直线回归关系(b=0.802,P<0.001); NF-kappa BP65蛋白表达和VCAM-1表达也呈直线回归关系(b=0.711,P<0.001). 结论 NF-kappa B的激活导致动脉粥样硬化患者体内ICAM-1和VCAM-1表达增强,加速了动脉粥样硬化病变过程.     

VEGF在白血病患者血清中的表达及意义

采用ELISA法测定40例白血病患者治疗前后血管内皮生长因子(VEGF)的表达,并与对照组进行比较,同时检测骨髓幼稚细胞水平,分析VEGF与白血病类型、病情及预后的关系.结果 显示,白血病患者的血清VEGF水平明显高于正常对照组(P<0.05),血清VEGF水平与白血病病情正相关.认为检测血清中VEGF的表达对白血病的诊断、治疗和预后的判断有一定意义.     

急性白血病骨髓中的血管生成的研究

目的 研究血管生成在急性白血病发病，预后中的作用。方法 应用免疫组化法检测30例初治急性白血病患者的骨髓活检组织治疗前后微血管密度（MVD）及血管内皮生长因子（VEGF）的变化。结果 初治急性白血病患者骨髓病理组织中的MVD和VEGF阳性率明显高于正常对照组；骨髓完全缓解后其MVD及VEGF阳性率明显下降，与正常对照组无显著性差异；治疗后未完全缓解组的MVD及VEGF阳性率较治疗前无显著性下降。结论 急性白血病骨髓中存在血管生成，可能与急性白血病的预后有关。     

急性白血病患者骨髓血管新生的研究及临床意义

目的　研究急性白血病 (AL)患者骨髓血管新生、骨髓液血管内皮生长因子 (VEGF)水平并探讨其临床意义。方法　采用免疫组化方法 ,以兔抗人Ⅷ相关抗原多克隆抗体标记骨髓内皮细胞 ,光学显微镜下计数骨髓全切片微血管数 ;用ELISA方法检测骨髓液VEGF水平。结果　骨髓微血管数在初发未治组 (2 2 82 /× 4 0 0视野 )明显高于正常对照组 (7 17/× 4 0 0视野 )及骨髓缓解组(8 5 7/× 4 0 0视野 ) (P值均 <0 0 0 1) ,而骨髓缓解组仍高于正常对照组 (P <0 0 5 ) ,初发未治组与复发难治组 (2 1 83/× 4 0 0视野 )之间的差异无显著性 (P >0 0 5 )。初发未治组中 ,骨髓微血管数在 2 3例急性淋巴细胞白血病 (ALL)患者 (2 3 0 9/× 4 0 0视野 )与 4 3例急性非淋巴细胞白血病 (ANLL)患者(2 2 37/× 4 0 0视野 )、FAB亚型M1～ 3 (2 2 91/× 4 0 0视野 )与M4～ 5患者 (2 1 4 6 /× 4 0 0视野 )之间的差异均无显著性 (P值均 >0 0 5 ) ,而骨髓微血管数与骨髓原始细胞百分数呈正相关性 (r =0 311,P <0 0 1)。骨髓液VEGF水平 ,在初发未治组 (188 88ng/L)显著高于骨髓缓解组 (78 74ng/L)和正常对照组 (79 5 2ng/L) (P值均 <0 0 1) ,而后两者之间的差异无显著性 (P >0 0 5 ) ;在ANLL组(2 70 12ng/L)显著高于ALL组 (1     

急性白血病患者骨髓MVD与血清 VEGF水平的研究

目的：探讨急性白血病(AL)骨髓血管新生的状态及意义。方法：应用改良的GMA塑料包埋骨髓病理切片进行免疫组化染色检测AL患者治疗前后骨髓微血管密度(MVD)，应用ELISA方法检测AL患者治疗前后血清血管内皮生长因子(VEGF)的浓度。结果：AL患者治疗前骨髓MVD明显增加，化疗缓解后骨髓MVD降至正常水平，复发者骨髓MVD再次升高至化疗前水平；AL患者治疗前血清VEGF水平比正常对照明显升高，化疗缓解的患者VEGF水平仍然较高。结论：骨髓血管新生在AL发生过程中有着重要的作用，血清VEGF水平能够在一定程度上解释白血病骨髓血管新生现象。     

Vascular endothelial growth factor and interleukin-6 in paracrine tumor-stromal cell interactions in multiple myeloma

Vascular endothelial growth factor (VEGF), a multifunctional cytokine, potently stimulates angiogenesis including tumor neovascularization. Although well established in solid tumors, the role of VEGF in bone marrow neoangiogenesis and paracrine tumor-stromal cell interactions in lymphohematopoietic malignancies has not been fully elucidated. In multiple myeloma (MM), marrow neovascularization parallels disease progression. This parallel prompted us to investigate the expression and secretion of VEGF by myeloma cells and its potential effects in myeloma-marrow stroma interactions. The biologically active splice variants VEGF165 and VEGF121 were expressed and secreted by myeloma cell lines and plasma cells isolated from the marrow of patients with MM. As shown by immunocytochemistry or RT-PCR, myeloma cells did not express or weakly expressed the VEGF receptors FLT-1 and FLK-1/KDR, indicating that autocrine stimulation is unlikely. In contrast, FLK-1/KDR was abundantly expressed by marrow stromal cells. Therefore, we studied the effects of VEGF on marrow stroma, focusing on the secretion of interleukin-6 (IL-6), a potent growth factor for myeloma cells and an inhibitor of plasma cell apoptosis. Exposure of stromal and microvascular endothelial cells to recombinant human (rh) VEGF165 or VEGF121 induced a time- and dose-dependent increase in IL-6 secretion (14- to 27-fold at 50 ng/mL after 24 hours, P <.001). Conversely, rhIL-6 stimulated VEGF expression and secretion in myeloma cell lines (40%-60%; P <.05) and to a variable degree (up to 5.3-fold; P <.005) in plasma cells purified from the marrow of patients with MM. This mutual stimulation suggests paracrine interactions between myeloma and marrow stromal cells triggered by VEGF and IL-6. (Blood. 2000;95:2630-2636)     

Role of the microenvironment in promoting angiogenesis in acute myeloid leukemia

Angiogenesis is a crucial event in the survival and progression of solid tumors. To determine whether angiogenesis in acute myeloid leukemia (AML) is an intrinsic property of leukemic cells, the vascularity of bone marrow biopsies was determined. Bone marrow vascularity in newly diagnosed or post-chemotherapy AML patients was increased 4-fold (P < 0.01) and 8.7-fold (P < 0.01), respectively, relative to controls. Vascular endothelial growth factor (VEGF) expression by AML blast cells was assessed by immunohistochemistry, and bone marrow cell supernatants were assayed for secretion of VEGF, fibroblast growth factor-2 (FGF-2), and endostatin by enzyme-linked immunosorbent assay. Diffuse cytoplasmic and strong extracellular VEGF immunoreactivity was seen in bone marrow aspirates from AML patients, but not controls. In contrast, there was no difference in the levels of VEGF, FGF-2, and endostatin secreted by mononuclear cells cultured from bone marrows of AML patients compared to normal controls following two days of culture in vitro. Total angiogenic potential of bone marrow cell supernatants was assessed by endothelial sprouting in vitro and by a chick chorioallantoic membrane assay. No differences were found between 2-day conditioned medium from normal and AML bone marrow mononuclear cells in either assay. Our data show a discrepancy between bone marrow vascularity and VEGF expression in vivo and VEGF expression and angiogenesis from 2-day conditioned medium ex vivo. This suggests that angiogenesis in AML likely represents a response to microenvironmental factors in vivo, rather than being an intrinsic property of leukemic cells.     

Absence of Clinical Prognostic Value of Vascular Endothelial Growth Factor and Microvessel Density in Multiple Myeloma

Vascular endothelial growth factor (VEGF) is considered a potent stimulator of angiogenesis. In multiple myeloma (MM), it has been reported that bone marrow angiogenesis parallels tumor progression and correlates with a poor prognosis. To investigate the role of angiogenesis in MM, we investigated VEGF expression and microvessel density (MVD) in the bone marrow of 75 MM patients by immunohistochemical methods. VEGF expression was observed in 87.3% (62 of 71) of patients. MVD was 69.42 +/- 9.67 (mean +/- SE) compared with the normal control values of 26.81 +/- 2.85. MVD values were 73.98 +/- 11.27 and 36.04 +/- 6.99 in the VEGF-positive and VEGF-negative groups, respectively. The MVD of patients in the VEGF-positive group was significantly higher than in the VEGF-negative group (P = .045). However, there were no significant differences in various clinical parameters, such as age, sex, hemoglobin, platelet count, serum levels of albumin, calcium, creatinine, and beta2-microglobulin, and bone marrow plasma cell percentage, between the VEGF-positive and VEGF-negative groups. Multivariate analysis revealed that age, hemoglobin, platelet count, serum levels of albumin and creatinine, and bone marrow plasma cell percentage were correlated with overall survival, whereas VEGF expression or MVD was not. In conclusion, our results suggest that VEGF is highly expressed and that MVD is increased in MM, indicating that angiogenesis may play a role in MM. Although MVD in the bone marrow of the VEGF-positive group is significantly higher compared with the VEGF-negative group (P = .045), VEGF is not correlated with overall survival. Further studies that include other angiogenic factors are needed to determine the functional role of angiogenesis in MM.     

急性髓系白血病及骨髓增生异常综合征患者血管内皮生长因子表达的研究

目的了解血管内皮生长因子(VEGF)在急性髓系白血病(AML)和骨髓增生异常综合征(MDS)中血管新生的作用。方法用ELISA法检测20例AML及24例MDS患者骨髓单个核细胞培养上清VEGF的表达。结果AML患者骨髓细胞VEGF的水平(267.35～412.30ng/L)高于对照组(128.17ng/L),化疗后完全缓解组的VEGF水平明显下降(P<0.05)化疗后未缓解组的VEGF水平与治疗前比较无显著下降(P>0.05)。MDS高危型(RAEB或RAEBt)中VEGF水平与低危型(RA或RAS)及对照组相比均明显升高(P<0.05),其中RAEBt与AML患者相比差异无显著性(P>0.05)。结论AML及高危型MDS患者骨髓中存在血管新生,VEGF在AML及MDS发病中起着重要作用,VEGF的表达与MDS病程进展有关。