肿瘤血管形成诱导和抑制因子研究进展

1肿瘤发生发展与血管形成肿瘤发生后生长过程大致可分为以下几个时期：第一时期为无血管期或称为血管前期。该期的肿瘤细胞主要靠弥散获得营养c如动物和人体内原位癌,浅表的皮肤黑色素瘤以及腹膜面小肿瘤,早期移植性小瘤灶等。第二时期为血管期也称为血管浸润性生长期。该时期瘤组织有新血管形成,这些血管不断地为痛组织提供营养和排除代谢产物,使肿瘤得以迅速增长。第三时期为转移期。当肿瘤生长到一定程度就向周围组织侵袭及发生转移,但转移灶形成过程,也会出现第一、二期的发展阶段。1．1肿瘤血管形成的实验研究一般利用不同移植物…     

肿瘤血管形成及其抑制因子研究进展

血管新生是从已形成的血管网络芽生出新的血管.肿瘤的生长、浸润和转移是一个依赖于血管的过程抑制新生血管的生成可使肿瘤细胞进入休眠状态并诱导死亡.     

肿瘤血管形成抑制因子研究进展

肿瘤血管形成受到生长因子和抑制因子的双重调节。本文对不含有ＥＬＲ－ｍｏｔｉｆ的α趋化因子、血管抑制素、内抑素、血小板反应素－１等血管形成抑制生长的来源、分子生物学和生物学功能作一综述。     

血管形成抑制因子HIAF-1在昆虫细胞中的高效表达和活性研究

背景与目的：大肠杆菌表达的人抑制血管生成因子－1（human inhibiting angiogenesis factor-1,HIAF-1）可抑制肿瘤血管的形成。本研究在昆虫细胞中表达HIAF－1,并研究其生物活性。方法：用DNA重组技术,构建了重组杆状病毒表达载体pAcuW51-HIAF-1,用Cellfectin将其与线性病毒DNA共转染昆虫细胞sf9,获得重组病毒。用SDS－PAGE电泳和Western blot鉴定重组病毒感染的昆虫细胞中重组HIAF－1重组蛋白的表达。重组蛋白经亲和层析纯化后,在体外用MTT法检测其对内皮细胞的作用,用食管癌移植瘤研究其体内抑瘤活性。结果：在昆虫细胞中高效表达了分子量为26kDa的HIAF－1重组蛋白,其表达量可达昆虫可溶性蛋白的5％－10％。重组HIAF－1蛋白不仅在体外显著抑制内皮细胞的生长,IC50值为3．1μg/ml,而且显著抑制食管癌移植瘤的生长。结论：在昆虫细胞中高效表达了分子量为26kDa的HIAF－1重组蛋白,其表达量可达昆虫可溶性蛋白的5％－10％。重组HIAF－1蛋白不仅在体外显著抑制内皮细胞的生长,IC50值为3．1μg/ml,而且显著抑制食管癌移植瘤的生长。结论：在昆虫细胞中高效表达了HIAF－1重组蛋白;HIAF－1重组蛋白的生物活性优于相应的原核重组蛋白。     

软骨源性血管形成抑制因子的研究进展

血管生成是肿瘤生成、入侵和转移的关键环节 ,原发肿瘤和转移性肿瘤持续生长的首要条件是诱导新血管的形成[1,2 ] 。因而抑制肿瘤的血管生成 ,切断肿瘤的营养供应及转移途径 ,从而抑制肿瘤的生长和转移 ,是一条很有希望的肿瘤治疗新途径[2 ] 。目前已获得多种血管形成抑制因子 ,如ＡＧＭ 14 70 ,干扰素等 ,有些已经应用于临床 ,但大部分由于毒性大而应用受限[3 ] 。软骨源性血管形成抑制因子由于活性高而无毒副作用 ,倍受瞩目。Ｄｕｐｏｎｔ[4] 发现 ,软骨对肿瘤组织的侵噬具有抵抗性。与软骨有关的肿瘤—软骨肉瘤 ,是血管化最轻的实体瘤。…     

血管生长抑制因子

恶性实体肿瘤的生长必须依赖新生血管持续不断地为其提供营养和排除代谢产物。因此，如能抑制或破坏这些新生血管就可防止恶性肿瘤的生长与转移，这是近年来癌症研究的一个新领域。国外为寻找效果好、毒性低的血管生长抑制因子进行了大量研究。血管生长抑制因子对癌症治疗具有良好前景。     

血管发生抑制因子 restin的克隆表达及其抗血管活性的初步研究

目的：克隆人血管发生抑制因子restin(hRs)，在E.coli中融合表达，并测定其抗血管活性。方法：用RT-PCR法从中国人胎盘组织中扩增hRs基因，重组人pGEM-T载体中并测定鉴定，构建原核表达载体pGEX-hRS，表达融合蛋白GST-hRS。融合蛋白经亲和纯化及凝血酶切后，采用鸡胚绒毛膜尿囊膜试验检测其抗血管生成活性。结果：RT-PCR产物为564bp，测序结果与Genbank中胶原XV(COL15A1)的C端序列一致，但在21位（TCT→TCG）引起丝氨酸的同义突变，82位（ACA→TCA）引起丝氨酸突变为苏氨酸。诱导表达的人GST-hRS融合蛋白经凝血酶切后，分子量为20kD，具有抗血管生成活性。结论：hRS的成功克隆、表达为抗血管生成治疗实体瘤的研究奠定了实验基础 。     

肿瘤血管形成因子的研究进展

肿瘤血管形成因子的研究进展王贵齐（综述）徐光炜（审阅血管形成即新的毛细血管芽发展过程是胚胎发育、肾脏、脑及其它器官血管化所必需的生理过程。成年人女性经期及伤口愈合时所发生的血管形成是短暂的且受机体严格的调节。与之相反，病理性血管形成则是以内皮细胞的持...     

血管生成抑制因子的研究进展

血管生成与肿瘤的生长、侵袭和转移密切相关,目前,以抑制血管生成为靶点治疗肿瘤有望成为抗肿瘤新疗法之一。近年来,研究发现了多种内源性血管生成抑制因子,其中一些已进入了临床试验阶段。本文对内源性血管生成抑制因子的结构、功能、作用机制及其在肿瘤治疗中的应用进行综述。     

血管生成抑制剂TNP—470抑制Lewis肺癌生长和转移的实验研究

目的采用Ｌｅｗｉｓ肺癌来验证血管生成抑制剂ＴＮＰ－４７０对肿瘤生长和转移的抑制作用。方法将Ｌｅｗｉｓ肺癌细胞接种于Ｃ５７ＢＬ小鼠皮下（２．４×１０６／鼠）。将２０只小鼠随机分成对照组和治疗组,自第２天起分别给予溶剂（３％酒精）０．２ｍｌ和ＴＮＰ－４７０（４０ｍｇ／ｋｇ）,隔天给药１次,共８次。第２２天时,测定对照组和治疗组的皮下瘤重及肺转移率,分别进行ｔ检验和χ２检验。结果两组皮下瘤重分别为３．７７±１．０５ｇ和１．９８±０．９６ｇ（Ｐ＝０．０００９）;两组肺转移率分别为８０％和３０％（Ｐ＝０．０３）。结论血管生成抑制剂ＴＮＰ－４７０能明显抑制Ｌｅｗｉｓ肺癌的生长和转移。     

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     

Overexpression of the p53-inducible brain-specific angiogenesis inhibitor 1 suppresses efficiently tumour angiogenesis

The brain-specific angiogenesis inhibitor 1 gene has been isolated in an attempt to find fragments with p53 "functional" binding sites. As reported herein and by others, brain-specific angiogenesis inhibitor 1 expression is present in some normal tissues, but is reduced or lost in tumour tissues. Such data and its particular structure prompted the hypothesis that brain-specific angiogenesis inhibitor 1 may act as a mediator in the local angiogenesis balance. We herein demonstrate that brain-specific angiogenesis inhibitor 1 over-expression suppresses tumour angiogenesis, delaying significantly the human tumour growth in immunodeficient mice. The inhibitory effect of brain-specific angiogenesis inhibitor 1 was documented using our intravital microscopy system, strongly implicating brain-specific angiogenesis inhibitor 1 as a mediator in the control of tumour angiogenesis. In contrast, in vitro tumour cell proliferation was not inhibited by brain-specific angiogenesis inhibitor 1 transfection, whereas some level of cytotoxicity was assessed for endothelial cells. Immunohistochemical analysis of tumour samples confirmed a reduction in the microvessel density index in brain-specific angiogenesis inhibitor 1-overexpressing tumours. At messenger level, moderate changes could be detected, involving the down-regulation of vascular endothelial growth factor and collagenase-1 expression. Furthermore, brain-specific angiogenesis inhibitor 1 expression that was lost in a selection of human cancer cell lines could be restored by wild-type p53 adenoviral transfection. Brain-specific angiogenesis inhibitor 1 should be considered for gene therapy and development of efficient drugs based on endogenous antiangiogenic molecules.     

Protein C inhibitor inhibits breast cancer cell growth, metastasis and angiogenesis independently of its protease inhibitory activity

Protein C inhibitor (PCI) regulates the anticoagulant protein C pathway and also inhibits urinary plasminogen activator (uPA), a mediator of tumor cell invasion. In the present study, we evaluated the effect of human PCI and its inactive derivatives on tumor growth and metastasis of human breast cancer (MDA-231) cells, and on angiogenesis in vivo. The invasiveness of MDA-231 cells was inhibited by recombinant intact PCI, but not by reactive site-modified PCI (R354APCI) or by the N-terminal fragment of protease-cleaved PCI (NTPCI). The in vitro invasiveness of MDA-231 cells expressing intact PCI (MDA-PCI) was significantly decreased as compared to MDA-231 cells expressing R354APCI (MDA-R354APCI) or NTPCI (MDA-NTPCI). Further, in vivo growth and metastatic potential of MDA-PCI, MDA-R354APCI and MDA-NTPCI cells in severe combined immunodeficient (SCID) mice were significantly decreased as compared to MDA-Mock cells. Angiogenesis was also significantly decreased in Matrigel implant containing MDA-PCI, MDA-R354APCI or MDA-NTPCI cells as compared to that containing MDA-Mock cells. In vivo angiogenesis in rat cornea and in vitro tube formation were also inhibited by recombinant intact PCI, R354APCI and NTPCI. Furthermore, the anti-angiogenic activity of PCI was strong as cleaved antithrombin (AT), and slightly stronger than that of plasminogen activator inhibitor (PAI)-1 and pigment epithelium-derived factor (PEDF). Overall, this study showed that, in addition to a reactive site-dependent mechanism, PCI may also regulate tumor growth and metastasis independently of its protease inhibitory activity by inhibiting angiogenesis.     

血管生成抑制剂YH-16抑制结肠癌肝转移的研究

背景与目的:YH-16是新合成的重组人血管内皮抑制素,Ⅱ期临床试验已证实YH-16联合化疗治疗晚期非小细胞肺癌具有协同作用。本文探讨血管生成抑制剂YH-16对结肠癌肝转移的抑制作用。方法:MTT法测定血管生成抑制剂YH-16对血管内皮细胞和结肠癌CT26细胞的IC50;经小鼠脾脏下极包膜下注入CT26细胞建立结肠癌肝转移模型,60只小鼠随机分为对照组、低剂量YH-16组、中剂量YH-16组、高剂量YH-16组,YH-16剂量分别为0mg/kg、0.40mg/kg、0.75mg/kg和1.50mg/kg,术后2周观察各组小鼠肝转移情况,采用免疫组化方法检测肝转移瘤组织中血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)的表达和肿瘤微血管密度(microvesseldensity,MVD)。结果:(1)YH-16对结肠癌CT26细胞和血管内皮细胞的IC50分别为(2.16±0.28)μg/ml和(0.64±0.10)μg/ml,前者是后者的3.38倍;(2)对照组,低、中、高剂量YH-16组肝转移率分别为100.0%、92.3%、80.0%和73.3%。高剂量YH-16组肝转移瘤数目明显低于对照组、低剂量YH-16组和中剂量YH-16组(P值均<0.05);(3)对照组,低、中、高剂量YH-16组脾脏肿瘤体积的中位数分别为1.180cm3、1.201cm3、0.887cm3和0.781cm3,四组比较无显著性差异(P>0.05);(4)YH-16各剂量组肝转移瘤组织中VEGF的表达较对照组无明显降低,四组肝转移瘤的MVD分别为65.00±9.58、58.15±8.81、51.60±7.10和44.53±11.47,中、高剂量YH-16组的MVD计数较对照组明显降低,高剂量YH-16组MVD计数较中剂量和低剂量YH-16组明显降低(P值均<0.05)。结论:血管生成抑制剂YH-16可以明显抑制结肠癌肝转移。     

抗肿瘤新生血管生成治疗的局限及应对策略

抗肿瘤新生血管治疗是近年来兴起的一种治疗肿瘤的新模式,已经在相关肿瘤领域确立了一线治疗地位.随着抗肿瘤新生血管生成治疗在临床的广泛应用也逐步显露出它的一些局限性如耐药、治疗窗不明确、缺乏有效的生物标记物.我们可通过联合化疗、合理规律的化疗方案、抗血管生成拟态(VM)治疗等策略加以克服.     

血管生成抑制剂TNP-470对小鼠肉瘤肺转移模型的影响

目的　建立肉瘤血行肺转移模型 ,探讨血管生成抑制剂TNP 470对肉瘤肺转移的形成及生长的影响。方法　筛选经反复小鼠腹腔注射 ,能持续引起小鼠血性腹水的S 1 80腹水癌细胞。按每鼠 1 .6× 1 0 6 / 0 .2ml经尾静脉接种于 1 5g左右的离乳幼鼠后随机分为对照组及TNP 470治疗组 ,每组 1 0只。接种后 2 4h起隔天皮下注射给药 ,第 1 5天处死小鼠 ,称肺湿重。肺组织切片 ,镜下计转移灶数。结果　经筛选的S 1 80肉瘤细胞尾静脉注射可成功建立小鼠血行肺转移模型。TNP 470 1 0 0mg/kg可有效减少小鼠肺转移模型中转移灶的数目并抑制转移灶的增大 ,但尚不能完全抑制转移灶的形成。结论　使用TNP 470等血管生成抑制剂可有效抑制肿瘤血行转移灶的形成和生长     

Inhibition of tumor growth and metastasis by angiogenesis inhibitor TNP-470 on breast cancer cell lines in vitro and in vivo

Antitumor and antimetastatic activity of the angiogenesis inhibitor O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), a semisynthetic analogue offumagillin, was evaluated in breast cancer cell lines. In an in vitro MTTassay, after 72 hrs continuous exposure to TNP-470, growth inhibition wasobserved in all seven cell lines of murine (JYG-A, JYG-B, DD-762, andBALB/c-MC) or human (KPL-1, MDA-MB-231, and MKL-F) origin, in which the50% inhibitory concentrations (IC₅₀) at 72 hrstreatment were 4.6, 4.4, 4.6, 10.1, 35.0, 25.3, and 33.4 µg/ml,respectively. In an in vivo assay using JYG-A, JYG-B, KPL-1, and MDA-MB-231cells by orthotopic (right thoracic mammary fat pad) transplantation infemale nude mice, TNP-470 at 30 or 50 mg/kg body weight was injected s.c.every other day from the day of tumor cell inoculation until the end of theexperiment. The inhibitory effect on primary tumor growth was obtained inall four cell lines in a dose-dependent manner. In the 50 mg/kgTNP-470-treated group, the reductions in tumor weight of the JYG-A, JYG-B,KPL-1, and MDA-MB-231 cells with respect to the controls were 50%,30%, 4%, and 49%, respectively. Metastasis was seen inthe JYG-A, JYG-B, and KPL-1 cells. The numbers of mice bearing pulmonarymetastases of JYG-A and JYG-B cells and regional axillary lymph nodemetastases of KPL-1 cells were reduced, and TNP-470 at the 50 mg/kg dose toKPL-1 cells significantly reduced lymph node metastases compared with thecontrol. Although the weight gain was retarded in the TNP-470-treated mice,weight loss was not seen. TNP-470 was highly effective in the treatment ofbreast cancer cells. These results suggest that the clinical use of TNP-470may be a promising treatment for breast cancer patients.     

小鼠Doc-1R基因的克隆及其表达

背景与目的：Doc-1R基因是1999年克隆的一种新的基因,已有的研究表明Doc-1R基因可能是一种潜在的抑癌基因。为了进一步研究此基因的功能,我们首先克隆了小鼠的Doc-1R基因,并对此基因的表达进行了初步的研究。方法;根据小鼠的Doc-1R基因cDNA序列,设计合成了基因组特异引物,应用巢式PCR对小鼠基因组步移文库进行扩增。对测序结果进行序列分析及剪接位点的鉴定。另外应用RT－PCR的方法研究了Doc-1R基因在小鼠肝,脾,胰,肾,肺,肠,心,脑,骨,肌肉,膀胱,卵巢,睾丸等13种组织的表达。结果：经二次基因组步移获得了小鼠Doc-1R基因组全长序列。基因组全长2787bp,含4个外显子和3个内含子,外显子与内含子接头符合GT／AG法则。RT－PCR结果表明,Doc-1R基因在小鼠13种组织和器官中均有表达。结论：成功克隆了小鼠Doc-1R基因,为进一步研究此基因的功能奠定了基础。RT－PCR表达结果提示此基因可能是对维持组织器官的功能具有重要的作用的管家基因。