内皮抑素基因治疗对小鼠肿瘤组织内新生血管形成抑制作用的实验研究

目的:研究腺病毒介导的内皮抑素基因治疗对小鼠肺肿瘤组织内部血管的抑制作用。方法:皮下注射2×106LLC细胞至C57BL/6小鼠背部建立小鼠肺癌种植瘤模型,2周后瘤内注射2×109pfu内皮抑素腺病毒载体治疗。免疫组化检测内皮抑素在肿瘤组织的原位表达,观察治疗对肿瘤生长的影响。免疫组化标记计数肿瘤内部血管密度,观察治疗对肿瘤血管的影响。结果:Admendostatin治疗从第2周起就可明显减缓肿瘤生长速度,P<0.05。免疫组化显示治疗组肿瘤组织内皮抑素蛋白强阳性表达,对照组阴性或痕量表达。Admendostatin治疗组、空载体对照组和阴性对照组肿瘤血管密度用CD31和CD105抗原标记分别为37.5±4.6、65.2±5.8、68.5±4.5和10.5±3.2、39.7±5.6、42.4±4.8。治疗组与两对照组相比差异有统计学意义,P<0.05;空载体对照与阴性对照相比差异无统计学意义,P>0.05。结论:腺病毒介导的内皮抑素基因治疗可抑制小鼠皮下种植瘤的生长,其作用机制是抑制肿瘤内部新生血管的生成,应用CD105抗原标记可准确反映新生血管的变化。     

增殖缺陷型腺病毒介导人内皮抑素基因抗乳腺癌的实验

 目的通过建立裸鼠乳腺癌肿瘤模型,在体内实验中进一步证实携带人内皮抑素基因的增殖缺陷型腺病毒Ad-hE的抗肿瘤作用。方法在BALB/c裸鼠皮下种植人乳腺癌细胞MCF-7,建立移植瘤模型。在瘤体内注射Ad-hE治疗,观察肿瘤生长,免疫组化检测肿瘤细胞内皮抑素的表达和计数肿瘤间质中微血管的数量。结果Ad-hE具有明显抑制肿瘤细胞生长的作用,瘤体内注射重组腺病毒后4周,Ad-hE治疗组肿瘤体积为(225.3±90.2)mm3,明显小于Ad-LacZ病毒对照组(794.9±189.8)mm3和空白对照组(890.7±102.5)mm3。免疫组化显示,乳腺癌细胞在感染腺病毒后能够有效表达内皮抑素,阳性细胞比率超过60%以上。Ad-hE治疗组瘤组织中血管数量(16.3±7.3)明显少于空白对照组(54.6±17.6)和Ad-LacZ病毒对照组(49.8±21.2)。结论增殖缺陷型腺病毒介导人内皮抑素的表达,具有明显抑制肿瘤细胞生长的作用。     

bFGF单克隆抗体抑制小鼠Lewis肺癌转移及血管新生

背景与目的:碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)是一种具有广泛生物学功能的细胞因子,在肺癌等多种肿瘤组织高表达,并参与其发生、发展进程.本研究旨在探讨bFGF单克隆抗体对C57 BL/6小鼠Lewis肺癌移植瘤生长、转移及血管新生的抑制作用.方法:采用皮下接种肿瘤细胞的方法,建立Lewis肺癌自发转移小鼠模型,随机分为PBS组、bFGF单克隆抗体MabF7治疗组,以及正常小鼠IgG阴性对照组,每组8只.自接种第9天开始给药,每3天1次,连续6次.同时观察Lewis肺癌小鼠健康状况,游标卡尺测量皮下移植瘤体积,给药6次后处死小鼠,称瘤质量,取肺组织计数各组小鼠肺表面转移瘤结节数,并用免疫组化法检测CD31的表达,以计算肿瘤组织微血管密度(microvessel density,MVD).结果:MabF7治疗组小鼠肿瘤体积与PBS组相比生长缓慢,小鼠瘤质量为(2.6±1.0)g,较PBS组的(5.1±1.3)g显著降低(P＜0.05);MabF7组小鼠肺表面转移瘤结节数为(3.0±2.1)个,显著低于PBS组的(12.3±2.4)个(P＜0.05).另外MabF7组可显著抑制肿瘤组织MVD表达水平.结论:bFGF单抗可明显抑制C57 BL/6小鼠Lewis肺癌生长及转移,其作用与抑制肿瘤增殖及微血管生成相关.     

小剂量奥沙利铂联合复方苦参注射液抗肿瘤血管生成的实验研究

目的:观察小剂量奥沙利铂(L-OHP)联合复方苦参注射液对小鼠皮下移植瘤生长的抑制作用以及对肿瘤组织中血管内皮生长因子(VEGF)的表达和微血管密度(MVD)的影响.方法:建立H22肝癌荷瘤小鼠模型后随机分为5组:5%葡萄糖注射液20ml/kg阴性对照组、环磷酰胺(CTX)20mg/kg阳性对照组、奥沙利铂0.75mg/kg组、复方苦参注射液3mL/kg组,复方苦参注射液3ml/kg+奥沙利铂0.75mg/kg联合组,每组10只.各组均给予腹腔等容量注射,连续用药10天.停药次日处死小鼠,计算瘤重抑制率.免疫组化法检测肿瘤组织中VEGF和MVD的表达.结果:小剂量奥沙利铂单药及联合复方苦参注射液组的瘤重均明显低于阴性对照组(P<0.05);肿瘤组织中的VEGF和MVD的表达明显低于阴性对照组(P<0.01).结论:小剂量奥沙利铂对H22肝癌细胞小鼠皮下移植瘤肿瘤组织具有明显抑制作用,联合复方苦参注射液具有明显相加抑制作用,能抑制肿瘤组织中VEGF和MVD的表达;小剂量奥沙利铂联合复方苦参注射液在体内可能具有抑制肿瘤组织血管生成的作用.     

罗勒多糖对肿瘤转移行为的影响

目的:研究腺病毒介导的鼠内皮抑素基因对胃癌的治疗作用.方法:利用病毒重组技术将内皮抑素克隆入增殖缺陷型腺病毒基因组中,观察体外转染表达后的生物学活性.通过建立荷人胃癌的裸鼠动物模型,分析基因转导后动物肿瘤细胞中内皮抑素的表达情况及对肿瘤的抑制作用.结果:构建了表达内皮抑素的重组腺病毒载体pCA13-mEndo;检测到内皮抑素在体外的mRNA水平和蛋白质水平均有表达;鸡胚绒毛尿囊膜实验表明其对血管生长起抑制作用;荷瘤裸鼠体内肿瘤生长抑制明显.结论:所构建的pCA13-mEndo重组腺病毒载体可有效表达具有生物学活性的内皮抑素,使得肿瘤内微血管生成减少,肿瘤细胞增殖减慢,为抗血管生成方法治疗实体瘤的临床应用奠定基础.     

口服VEGFR2 DNA疫苗抗小鼠Lewis肺癌血管生成的实验研究

背景与目的血管内皮生长因子(VEGF)及其主要受体血管内皮生长因子受体-2(VEGFR2)在肿瘤新生血管和肿瘤基质形成过程中起着重要作用。本研究的目的是观察口服VEGFR2 DNA疫苗抗C57BL/6小鼠Lewis肺癌皮下移植瘤生长的作用,并探讨其可能的作用机制。方法将重组DNA疫苗对小鼠进行免疫,通过观察小鼠Lewis肺癌皮下移植瘤大小,记录各组小鼠离体肿瘤湿重,检测移植瘤微血管密度(MVD)及血液CD3 、CD8 T细胞水平,评价重组疫苗的抑瘤作用。结果疫苗组、空质粒组、生理盐水组的MVD分别为1.75±1.07、6.89±2.52、7.57±3.75,肿瘤湿重分别为(2.05±1.32)、(4.83±1.47)、(5.12±1.02)g,疫苗组与其它两组比较,差异均有统计学意义(P<0.05)。接种肿瘤后,疫苗组CD3 T细胞仍维持较高水平,其它两组明显下降(P<0.05);疫苗组CD8 T细胞水平较其它两组明显升高(P<0.05)。结论口服VEGFR2DNA疫苗对小鼠Lewis肺癌皮下移植瘤的生长具有较强的抑制作用。该疫苗可能是通过杀伤肿瘤内皮细胞、抑制血管生成而起到抗肿瘤生长的作用。     

小剂量奥沙利铂联合复方苦参注射液抗肿瘤血管生成的实验研究

目的：观察小剂量奥沙利铂（L-OHP）联合复方苦参注射液对小鼠皮下移植瘤生长的抑制作用以及对肿瘤组织中血管内皮生长因子（VEGF）的表达和微血管密度（MVD）的影响。方法：建立H22肝癌荷瘤小鼠模型后随机分为5组：5%葡萄糖注射液20ml/kg阴性对照组、环磷酰胺（CTX）20mg/kg阳性对照组、奥沙利铂0.75mg/kg组、复方苦参注射液3ml/kg组,复方苦参注射液3ml/kg＋奥沙利铂0.75mg/kg联合组,每组10只。各组均给予腹腔等容量注射,连续用药10天。停药次日处死小鼠,计算瘤重抑制率。免疫组化法检测肿瘤组织中VEGF和MVD的表达。结果：小剂量奥沙利铂单药及联合复方苦参注射液组的瘤重均明显低于阴性对照组（P〈0.05）;肿瘤组织中的VEGF和MVD的表达明显低于阴性对照组（P〈0.01）。结论：小剂量奥沙利铂对H22肝癌细胞小鼠皮下移植瘤肿瘤组织具有明显抑制作用,联合复方苦参注射液具有明显相加抑制作用,能抑制肿瘤组织中VEGF和MVD的表达;小剂量奥沙利铂联合复方苦参注射液在体内可能具有抑制肿瘤组织血管生成的作用。     

小剂量希罗达抗小鼠结肠癌生长及微血管的形成

 目的 研究小剂量希罗达对小鼠结肠癌CT 26移植瘤生长和微血管生成的影响。 方法 建立小鼠结肠癌皮下移植瘤模型,随机分组：治疗组持续小剂量希罗达90mg/kg（每只0.1ml）灌胃,对照组给予相应体积生理盐水。给药后观察每组小鼠肿瘤生长情况,肿瘤长出后隔天测量肿瘤体积,接种肿瘤细胞两周后,处死全部小鼠,免疫组织化学法检测肿瘤组织血管内皮生长因子（VEGF）表达及肿瘤组织微血管密度（MVD）,Western blot法检测肿瘤组织VEGF的表达。 结果 治疗组肿瘤生长受到明显抑制,治疗组VEGF表达强度及MVD低于对照组,VEGF与MVD间呈正相关;Western blot检测治疗组VEGF表达明显减弱。实验结束时未见明显不良反应。 结论 小剂量希罗达能通过抑制肿瘤微血管生成发挥抗肿瘤作用,其机制与抑制VEGF的表达有关。     

VEGF-C基因沉默对裸鼠移植乳腺癌淋巴管和血管生成影响的观察

目的:探讨血管内皮生长因子C(VEGF-C)基因沉默对裸鼠移植乳腺癌淋巴管和血管生成的影响.方法:构建VEGF-C小分子干扰RNA(VEGF-C/SiRNA)慢病毒载体,转染靶细胞MCF-7,Real-time PCR检测VEGF-C敲减效率,将其接种于裸小鼠背部皮下(KD组),并设置MCF-7细胞对照组(CON组)及空载体对照组(NC组),观察裸小鼠的成瘤情况,通过免疫组化检测VEGF-C表达、微血管密度(MVD)及微淋巴管密度(LVD).结果:VEGF-C/SiRNA转染MCF-7细胞后,与对照组相比,VEGF-C表达显著降低,敲减效率为49.2％,P=0.017.接种后各组细胞均能成瘤,KD组6只,NC组7只,CON组8只,成瘤情况差异无统计学意义,P=0.631.HE染色结果显示,KD组可见肿瘤细胞明显减少,出现较多凋亡和坏死细胞.免疫组化检测显示,CON组VEGF-C蛋白表达(+)1只,(++)1只,(+++)4只:NC组表达(++)1只,(+++)5只;KD组表达(+)5只,(++)1只,KD组移植瘤VEGF-C蛋白表达显著低于对照组,P=0.02.肿瘤组织内LVD计数,KD组5.33±1.63,CON组8.50士1.05,NC组8.17±1.83,KD组显著低于NC组和CON组,差异有统计学意义,P=0.005;MVD计数,KD组8.83±1.72,CON组11.02±1.41,NC组11.17±2.48,KD组MVD略低于CON组及NC组,但差异无统计学意义,P=0.097.结论:慢病毒介导的VEGF-C/siRNA可有效敲减VEGF-C表达,显著减少乳腺癌组织的淋巴管生成,但对血管生成无显著影响.     

重组人血管内皮抑素联合重组人p53腺病毒治疗乳腺癌的实验性研究

背景与目的目前多基因、多靶点联合治疗已成为肿瘤研究的方向之一,本文探讨重组人血管内皮抑素(endostatin,ES)联合重组人p53腺病毒(rAd/p53)对乳腺癌裸鼠移植瘤的抑制作用.方法建立人乳腺癌裸鼠模型,成瘤后随机分为对照组、ES组、rAd/p53组和联合组,分别给予处理;观察裸鼠的一般情况、体重和肿瘤的体积,并检测肿瘤组织微血管密度(MVD,microvessel density)和血管内皮生长因子(VEGF,vascular endothelial growth factor)的表达.结果成瘤后ES组,rAd/p53组和联合组抑瘤率分别为1.75%、43.36%和58.68%,rAd/p53组和联合组可明显抑制肿瘤生长(P＜0.05),联合组抑瘤作用更为显著;对照组、ES组、rAd/p53组与联合组肿瘤组织的MVD及VEGF表达H-评分分别为(30±5、24±4、24±4、18±4)和(46±8、39±8、40±5、29±5),联合组均低于其余各组,差异有显著性(P＜0.05);各组均未见不良反应,体重变化无显著性(P＞0.05).结论重组人血管内皮抑素联合rAd/p53可显著抑制裸鼠乳腺癌的生长及微血管生成.     

Intratumoral administration of endostatin plasmid inhibits vascular growth and perfusion in MCa-4 murine mammary carcinomas

Endostatin, a fragment of the COOH-terminal domain of mouse collagen XVIII is a recently demonstrated endogenous inhibitor of tumor angiogenesis and endothelial cell growth. Antiangiogenic therapy with endostatin in animals requires multiple and prolonged administration of the protein. Gene therapy could provide an alternative approach to continuous local delivery of this antiangiogenic factor in vivo. Established MCa-4 murine mammary carcinomas, grown in immunodeficient mice, were treated with intratumoral injection of endostatin plasmid at 7-day intervals. At the time of sacrifice, 14 days after the first injection, endostatin-treated tumor weights were 51% of controls (P < 0.01). Tumor growth inhibition was accompanied by a marked reduction in total vascular density. Specifically, computerized image analysis showed a 18-21% increase in the median distances between tumor cells and both the nearest anatomical (CD31-stained) vessel [48.1 +/- 3.8 versus 38.3 +/- 1.6 microm (P < 0.05)] and the nearest tumor-specific (CD105-stained) vessel [48.5 +/- 1.5 versus 39.8 +/- 1.5 microm (P < 0.01)]. An increased apoptotic index of tumor cells in endostatin-treated tumors [3.2 +/- 0.5% versus 1.9 +/- 0.3% (P < 0.05)] was observed in conjunction with a significant decrease in tumor perfused vessels (DiOC7 staining), and an increase in tumor cell hypoxia (EF5 staining). Hypoxia resulting from endostatin therapy most likely caused a compensatory increase of in situ vascular endothelial growth factor (VEGF) and VEGF receptor mRNA expression. Increased immunoreactivity of endostatin staining in endostatin-treated tumors was also associated with an increased thrombospondin-1 staining [1.12 +/- 0.16 versus 2.44 +/- 0.35]. Our data suggest that intratumoral delivery of the endostatin gene efficiently suppresses murine mammary carcinoma growth and support the potential utility of the endostatin gene for cancer therapy.     

瘤内注射pSTbAT-脂质体复合物对裸鼠移植瘤抑制效应的实验研究

目的:构建血管生成抑制因子arresten基 因的真核表达载体,并观察瘤内注射质粒DNA 脂质 体复合物对裸鼠移植瘤生长的影响。方法:利用 PCR方法,由重组质粒pGEM Arr中扩增出基因;将 该基因定向克隆于真核表达载体中。20只裸鼠皮 下注射HepG 2细胞,成瘤后以成功构建的质粒 DNA 脂质体复合物瘤内注射,治疗后采用免疫组化 方法检测肿瘤血管密度,并利用arresten基因瘤内注 射的方法对抑制肿瘤的效果进行分析。结果:我们 成功构建arresten基因真核表达载体(pSTbAT)。实 验组肿瘤生长速度较对照组慢(实验组平均15.3个 阳性血管/10个视野,对照组平均112.5个阳性血 管/10个视野)。结论:瘤内注射arresten基因质粒 DNA复合物能抑制裸鼠移植瘤的血管生成,并能抑 制移植瘤的生长。     

原发性肝癌肿瘤血管密度及其表达的临床病理意义

目的研究肿瘤微血管密度（ＭＶＤ）和肿瘤血管生长因子（ＶＥＧＦ）的表达水平在原发性肝癌中的临床病理意义。方法对６３例手术切除的小肝癌（直径＜５ｃｍ）患者的临床病理资料进行了回顾性的研究。其中２年内转移复发组２９例,２年内无转移复发组３４例。每例取３张连续的切片分别进行ＨＥ染色、ＶＥＧＦ和生物素标记的荆豆凝集素Ｉ（Ｂｉｏ－ＵＥＡ－Ｉ）的免疫组化染色。根据Ｂｉｏ－ＵＥＡ－Ｉ染色的血管内皮细胞计数来测定ＭＶＤ。结果转移复发组和无转移复发组的ＭＶＤ分别为４９．６±２９．７、２２．７±２８．２（Ｐ＜０．０１）;转移复发组和无转移复发组的ＶＥＧＦ阳性率分别为８６．２％（２５／２９）、４７．１％（１６／３４）,两者差异有显著性（Ｐ＜０．０１）。两组在肿瘤分期、有无卫星灶、有无门脉癌栓３方面差异也具有显著性（Ｐ＜０．０５,Ｐ＜０．０５,Ｐ＜０．０１）。结论除肿瘤分期、卫星灶、门脉癌栓具有预后意义外,肝癌组织中ＭＶＤ和ＶＥＧＦ的表达也具有预后价值     

Endostatin对结肠癌生长和转移抑制作用的实验研究

背景与目的:结肠癌的生长、转移是血管生成依赖性的,血管生成抑制剂有望通过抑制肿瘤血管生成,诱导结肠癌细胞凋亡,有效地抑制结直肠癌的生长和转移。肿瘤的抗血管生成治疗对选择手术时机和方式、制定综合治疗方案,提高结肠癌患者5年生存率都具有重要意义。本实验研究血管生成抑制因子Endostatin对结肠癌生长和转移的抑制作用,并探讨其对结肠癌细胞凋亡的影响。方法:采用人结肠腺癌细胞株SW1116完整组织块裸鼠原位种植,建立类似于临床的结肠癌转移裸鼠模型。种植后第1周开始皮下注射Endostatin,每天一次,剂量为0mg/kg(对照组)、5mg/kg、10mg/kg、20mg/kg(治疗组),共用6周。种植后第7周处死动物,测量原位肿瘤瘤重、抑瘤率、肿瘤微血管密度(intratumoralmicrovesseldensity,MVD)、肿瘤细胞凋亡指数(apoptoticindex,AI),观察肿瘤细胞腹膜、肝、其他脏器转移及腹水情况。结果:Endostatin剂量为0mg/kg、5mg/kg、10mg/kg、20mg/kg时,原位肿瘤瘤重分别为(1.31±0.36)g、(0.42±0.17)g、(0.21±0.09)g、(0.13±0.05)g;抑瘤率分别为0%、67.9%、84.0%、90.1%;MVD分别为(12.8±4.1)、(5.9±2.5)、(2.2±1.4)、(0.74±O.3);AI分别为(3.87±2.61)%、(6.89±5.18)%、(13.24±4.76)%、(20.97±9.04)%;腹膜转移率分别为9     

Electrogene therapy using endostatin, with or without suicide gene therapy, suppresses murine mammary tumor growth and metastasis

Syngeneic inoculated metastatic mammary cancers received direct intratumoral injection of a plasmid vector containing either endostatin (pEndo) with or without a suicide gene (pHSVtk), pHSVtk alone or control vector once a week for 8 weeks. We applied electrogene transfer to the tumors after each injection and administered ganciclovir (GCV) to pHSVtk-transfected mice using an osmotic minipump. Anticancer efficacy was monitored using a variety of parameters, namely tumor volume, intratumoral microvessel density and DNA synthesis, number of mice with metastasis, and number of sites of metastasis per mouse. Tumor volume was significantly lower in all therapeutic groups, with the most effective growth suppression in the pEndo+pHSVtk/GCV group. Lymph node metastasis was significantly less frequent in all therapeutic groups, whereas the multiplicity of lung metastases was significantly lower only in the pEndo and pEndo+pHSVtk/GCV groups. All therapeutic groups showed significantly lower intratumor microvessel density and DNA synthesis. The pEndo and pEndo+pHSVtk/GCV groups also showed a significant reduction in the numbers of dilated lymphatic vessels containing intralumenal tumor cells. Our data suggest that endostatin electrogene therapy alone or in combination with pHSVtk/GCV suicide gene therapy is more beneficial than suicide gene therapy alone. The observed antimetastatic activity of endostatin may be of high clinical significance in the treatment of metastatic breast cancer.     

胃癌细胞异位分泌人绒膜促性腺激素对微血管密度的影响

目的研究胃癌组织中人绒膜促性腺激素（ＨＣＧ）对微血管密度（ＭＶＤ）的影响。方法运用ＳＰ法,对４０例胃癌手术切除标本进行抗人β促绒膜性腺激素（βＨＣＧ）多克隆抗体和抗因子ＦⅧ相关抗原抗体（ＦⅧＲＡｇ）免疫组织化学染色,观察癌组织、癌旁、正常组织各区域βＨＣＧ的表达和ＭＶＤ。结果所有胃癌组织中,βＨＣＧ仅在癌细胞内阳性表达,表达率为３７．５％（１５／４０）。ＭＶＤ在癌组织为２７．４±７．１,癌旁为１１．９±５．４,正常组织为４．８±１．５（Ｐ＜０．０５～０．０１）。βＨＣＧ阳性组的ＭＶＤ明显高于βＨＣＧ阴性组（Ｐ＜０．０５）,βＨＣＧ,ＭＶＤ和组织类型无关（Ｐ＞０．０５）,但βＨＣＧ和淋巴结转移有关（Ｐ＜０．０５）,伴有血道转移组的βＨＣＧ,ＭＶＤ均高于不伴血道转移组（Ｐ＜０．０５）。结论微血管生长与肿瘤的生长,血道转移有关,而ＨＣＧ对肿瘤的微血管有一定的促生长作用。     

Therapy of hematogenous melanoma brain metastases with endostatin.

PURPOSE: Cerebral metastases represent the most common type of brain tumors. This study investigated the effects of endogenous endostatin on hematogenous cerebral melanoma metastases. EXPERIMENTAL DESIGN: Murine K1735 melanoma cells were transfected with the mouse endostatin cDNA. Experimental tumors were induced either by s.c. injection, intracerebral implantation, or via injection into the internal carotid artery to simulate hematogenous metastatic spread. The effects of endostatin expression on tumor incidence, growth pattern, and vascularity were analyzed. RESULTS: In vitro secretion of endostatin by 2.5 x 10(5) cells within 24 hours was 0.12 +/- 0.03 ng, 4.35 +/- 0.4, and 1.18 +/- 0.7 ng/mL for wild type and two endostatin-transfected K1735 clones termed K1735-endo/2 and K1735-endo/8, respectively. Tumor inhibition in vivo correlated with endogenous endostatin production. Within 25 days, growth of s.c. K1735-endo/2 tumors was <20% compared with wild-type controls. Following intracerebral implantation the average survival time of mice was 27.8 +/- 2.6 versus 13.3 +/- 3.7 days in the K1735-endo/2 versus the wild-type group, respectively. Intracarotid injection of 1 x 10(5) wild-type cells killed the mice within 24 +/- 1.8 days. In contrast, endostatin expression prevented macroscopic metastatic tumor growth in 11 of 12 mice, although viable microscopic tumor pockets were detectable in all animals. CONCLUSION: Endostatin inhibits tumor progression of multiple cerebral metastases in vivo. Hematogenous micrometastases are more efficiently suppressed than tumors resulting from high focal cell numbers which may be due to a higher angiogenic signaling exerted by massive cell deposits. Endostatin may prevent solid tumor growth more effectively by inhibition of early angiogenesis.     

血管内皮生长因子在大肠癌中的表达及临床意义

目的研究肿瘤生长因子(VEGF)和肿瘤微血管密度(MVD)的表达水平在大肠癌中的临床意义.方法对86例手术切除的大肠癌患者的临床病理资料进行研究,并采用免疫组化方法检测VEGF蛋白和MVD在大肠癌的表达水平.结果VEGF表达阳性率为62.8%.MVD和VEGF表达的程度明显相关.VEGF的阳性表达与组织学类型及肿瘤位置无显著相关(P＞0.05),而与淋巴结转移、肝肺等远处转移及浸润深度有明显相关性(P＜0.05).结论VEGF是由肿瘤细胞所分泌的,并在大肠癌的生长和转移过程中发挥重要的作用.     

Angiogenesis and Lymphangiogenesis in Salivary Gland Adenoid Cystic Carcinoma and Mucoepidermoid Carcinoma

Background and Purpose: This study aimed to investigate the immunohistochemical expression of CD31 and podoplanin in order to examine angiogenesis and lymphangiogenesis, respectively in common malignant tumors of salivary glands. Materials and Methods: Forty formalin-fixed, paraffinated blocks (20 adenoid cystic carcinoma and 20 mucoepidermoid carcinoma blocks) were selected from the medical archives of Amir A’lam Hospital of Tehran, Iran. Sections from the blocks were stained by CD31 and D2-40 markers via immunohistochemistry. Clinical and demographic information was extracted from the patients’ records. Findings: There was a significant difference between tumors in terms of intratumoral microvessel density (MVD) (P< 0.001), total MVD (P< 0.001), and intratumoral lymphatic vessel density (LVD) (P= 0.011). In mucoepidermoid carcinoma, intratumoral MVD and LVD were greater than peritumoral MVD and LVD (P= 0.001 and P< 0.001, respectively). In mucoepidermoid carcinoma, there was no relationship between histological grade with MVD (total, intratumoral or peritumoral) or LVD (total, intratumoral or peritumoral) (P> 0.05). A similar finding was reported with respect to the histopathological grade of adenoid cystic carcinoma (P> 0.05). Conclusion: The higher level of angiogenesis and lymphangiogenesis in mucoepidermoid carcinoma, specifically at the center of tumor, compared to adenoid cystic carcinoma, may be attributed to differences in the clinical behaviors and metastasis of tumors. Moreover, considering the high LVD at the center of tumor in mucoepidermoid carcinoma and infrequency of metastasis to regional lymph nodes in adenoid cystic carcinoma, it can play a significant role in metastasis to regional lymph nodes.     

血管内皮抑制素对Calu-6裸鼠移植瘤的生物学效应

目的 探讨血管内皮抑制素对Calu-6裸鼠移植瘤生长及新生血管形成的影响.方法 在荷瘤裸鼠皮下注射不同剂量的血管内皮抑制素,观察注射后肿瘤体积的变化;应用免疫组化SABC法检测肿瘤组织中血管内皮生长因子(VEGF)、生存素(survivin)和环氧化酶-2(COX-2)蛋白的表达以及微血管密度(MVD)的变化;流式细胞术检测循环血管内皮细胞(CECs)的含量;应用逆转录聚合酶链反应(RT-PCR)和实时定量PCR检测外周血中CD146和CD105 mRNA的表达.结果经血管内皮抑制素治疗后,荷瘤鼠肿瘤体积明显减小;肿瘤组织中VEGF、survivin和COX-2蛋白的表达以及MVD均下降,且各治疗组与阳性对照组间的差异均有统计学意义(均P＜0.05);外周血中CECs、CD146和CD105 mRNA的含量均明显下降;活化CECs的含量与肿瘤组织中survivin和VEGF的表达以及MVD的变化均呈正相关.结论 血管内皮抑制素可通过下调移植瘤中VEGF、survivin和COX.2蛋白的表达以及减少MVD抑制肿瘤生长;活化CECs将可能作为理想的预测抗血管形成治疗预后的标记物应用于临床.