TNP-470对胃癌生长抑制作用的实验研究

目的研究TNP-470(血管生成抑制剂)对胃癌生长的抑制作用。方法采用裸鼠皮下种植人胃癌细胞株,成瘤后随机分为治疗组及对照组,皮下注射TNP-470 30mg/kg,隔日1次,连续8次后,处死动物,测定瘤重及肿瘤微血管密度。结果实验组瘤重明显低于对照组(P<0.05),肿瘤微血管密度明显低于对照组(P<0.05)。结论TNP-470可明显抑制胃癌细胞生长,治疗组肿瘤微血管密度明显低于对照组。     

三氧化二砷对裸鼠结肠癌腹腔转移的抑制作用

目的　研究Ａｓ２Ｏ３ 对裸鼠结肠癌腹腔转移的影响及其作用机制。方法　ＢＡＬＢ／Ｃｎｕ／ｎｕ裸鼠腹腔内接种结肠癌Ｌｏｖｏ 细胞后,随机分为５ 组,分别腹腔内注射生理盐水、表阿霉素及不同剂量的Ａｓ２Ｏ３ ,观察各组的致瘤率、腹水生成及生存时间。结果　表阿霉素、低剂量Ａｓ２Ｏ３ 与对照组相比可明显抑制裸鼠腹水的生长,延长生存期（ Ｐ＜ ０．０１） ;中高剂量Ａｓ２Ｏ３ 除上述作用外还可通过诱导肿瘤细胞凋亡消除腹腔内肿瘤细胞,抑制腹水生成,并使裸鼠寿命明显延长。结论　Ａｓ２Ｏ３ 可诱导结肠癌细胞凋亡,抑制或消除裸鼠肿瘤性腹水的生成,不同程度地延长生存期。     

TNP-470对Wistar大鼠C6胶质瘤细胞抑制作用的体内实验研究

目的探讨TNP-470对Wistar大鼠C6胶质瘤细胞的抑制作用。方法培养C6细胞，建立动物荷瘤模型。荷瘤后将Wistar大鼠随机分为肿瘤对照组（0．9％氯化钠注射液30mg,／kg隔日1次，共6次皮下注射）、TNP-470组（TNP-47030mg,／kg隔日1次，共6次皮下注射）、顺铂组（顺铂5mg／kg每周2次，共4次腹腔内注射）、联合给药组（TNP-47015mg,／kg隔日皮下注射，共6次；同时顺铂2．5mg／kg腹腔内注射，每周2次，共4次），每组15只。观察不同的药物对肿瘤生长的影响。结果与对照组相比，TNP-470治疗组肿瘤重量及最大横径显著降低（P〈0．001）；TNP-470治疗组及联合给药组G0／G1期细胞数显著高于对照组（P〈0．01），而TNP-470治疗组G0／G1期细胞数明显高于联合治疗组（P〈0．01）；TNP-470治疗组、联合给药组及顺铂治疗组G2／M期、S期细胞数显著低于对照组（P〈0．01）。与肿瘤对照组相比，联合给药组和顺铂治疗组及TNP-70组胶质瘤细胞凋亡百分率显著升高（P〈0．01）TNP-470治疗组胶质瘤细胞凋亡百分率显著高于联合治疗组（P〈0．01）。结论TNP-470是一种对胶质瘤有明显疗效的化疗药物，可明显抑制肿瘤生长，使肿瘤重量减轻，同时对大鼠本身体重影响小，行为学症状改变出现较少，疗效明显优于顺铂。     

氧化砷对人结肠癌裸鼠腹腔种植的抑制作用

目的 :研究氧化砷 (As2 O3)对结肠癌裸鼠腹腔转移的影响及其作用机制。方法 :建立BALB/C nu/nu裸鼠SW 480结肠癌腹水型移植瘤模型 ,腹腔内注射As2 O3,观察As2 O3对荷瘤鼠的抗癌作用。结果 :5 氟脲嘧啶 (5 FU)、低剂量As2 O3和高剂量As2 O3均可明显延长腹水型荷瘤鼠的生存时间 ,其延命率分别为 40 .1± 7.83 %、47.2 2± 8.69%和 92 .1 0± 1 4 .2 % ,高剂量As2 O3延命率明显高于 5 FU ,两组比较差异有显著性 (P <0 .0 1 )。治疗组腹水CEA水平明显低于阴性对照组。在普通光学显微镜下观察腹水标本 ,部分结肠癌细胞发生了较典型的细胞凋亡形态学改变。腹水细胞流式细胞仪检测可见凋亡峰。低剂量As2 O3凋亡率为 8.36± 2 .1 4 % ,高剂量As2 O3凋亡率为 1 5 .3± 3 .82 % ,均明显高于生理盐水组的 2 .40± 1 .1 8% (P <0 .0 1 )。结论 :As2 O3对结肠癌荷瘤鼠具有延长生命的作用 ,其作用机制主要是诱导结肠癌细胞凋亡。     

TNP-470对肺腺癌细胞增殖和凋亡的影响

目的 探讨TNP－470对肺腺癌细胞增殖和凋亡的影响及其作用机制。方法 以不同浓度的TNP－470信息处理地培养的肺腺癌AGZY－82A细胞株,以免疫组化S－P法检测肺腺癌细胞增殖核抗原（PCNA）、p53、bcl-2、血管内皮生长因子（VEGF）和VEGF受体Flk-1的表达。结果 随着TNP－470浓度的增加,肺腺癌细胞VEGF、Flk-1、PCNA的表达减少,p53、bcl-2的表达逐渐增加;当TNP－470为10^7μg/L时,细胞增殖几乎停止,随着TNP－470（10^4μg/L）作用时间的延长,肺腺癌细胞VEGF、Flk-1、PCNA的表达逐渐减少,而p53、bcl-2的表达逐渐增加。结论 TNP－470通过使肺腺癌细胞自分泌VEGF减少,Flk表达降低,从而抑制肺腺癌细胞增殖,促进其凋亡。     

TNP-470对小鼠Lewis肺癌的作用及其机制

TNP 4 70是烟曲霉素的衍生物O (氯乙酰 氨酰基 )烟曲霉醇 ,有高选择性抗血管新生作用。近年研究表明 ,应用血管新生抑制剂治疗肿瘤取得了明显的疗效 ,从而为肿瘤的治疗开辟了一条新的途径。我们利用复制荷Lewis肺癌小鼠模型 ,应用TNP 4 70 ,与常规化疗药物阿霉素 (ADM)联合或对照 ,观察其对原发和转移性肺癌的疗效。材料与方法　TNP 4 70由日本武田制药株式会社惠赠。ADM为汕头经济特区明治医药有限公司制造 ,批号 990 4 0 1。1.动物模型的复制及分组 :雄性纯系C5 7BL/ 6小鼠 (中国医科大学实验动物部提供 ) 4 0只 ,皮下接种Lewis…     

裸鼠荷人卵巢癌腹水瘤模型的构建

目的探讨建立裸鼠荷人卵巢癌腹水瘤模型的方法。方法将体外培养的人卵巢癌HO-8910细胞,调制成浓度分别为2．5×10^6、5×10^6、1．0×10^7、2．5×10^7、5×10^7／ml的细胞悬液,采用腹腔注射的方式接种至裸鼠体内,注射体积为200μl,接种后观察各组成瘤时间及成瘤率。结果成功地构建了人卵巢癌上皮性细胞HO-8910裸鼠腹水瘤模型。接种剂量为1．0×10^7、5．0×10^6和2．0×10^6个癌细胞者成瘤率分别为100％、100％、90％,成瘤时间分别在15～21d、18～24d、21～25d,腹水瘤形成后生存时间分别为10～15d、12～18d、28～35d。接种剂量为5．0×10^5、1．0×10^6个者30～42d形成腹水瘤,且成瘤率低（≤50％）。结论腹腔注入2．0×10^6个HO-8910是建立裸鼠荷人卵巢癌腹水瘤模型的好方法。     

肠复康合剂对裸鼠移植性人结肠癌血管生成的抑制作用

目的 :探讨肠复康合剂对裸鼠移植性人结肠癌 HT- 2 9血管生成的机制。方法 :将 2 4只裸小鼠随机分为模型组、肠复康组及西药组 ,每组 8只。各组分别于建立人结肠癌 HT- 2 9癌细胞株裸小鼠皮下移植瘤模型后 6 d给予相应药物 ,免疫组化染色结合图像分析系统半定量检测移植瘤组织微血管密度 (MVD)、血管内皮生长因子(VEGF)及其受体 (FL K- 1)、内皮抑素 (ES)的积分光密度 (IOD) ,计算 VEGF/ES比值 ,并使用逐步引入剔出模型进行单因素和多因素线性回归分析。结果 :与模型组相比 ,肠复康组可使移植瘤 MVD、VEGF、FL K- 1及 VEGF/ES比值显著降低 (P <0 .0 1) ,同时使 ES含量明显增高 (P <0 .0 1) ;西药组可明显降低瘤组织 VEGF、FL K- 1含量 (P<0 .0 1) ,但不能使移植瘤 MVD、ES和 VEGF/ES比值减少 (P >0 .0 5 )。单因素回归分析显示 ,瘤组织 VEGF、ES和 VEGF/ES比值与移植瘤 MVD相关 ,多因素回归分析显示 ,移植瘤 MVD与 VEGF/ES比值呈明显正相关 (r=0 .76 2 ,P <0 .0 1)。结论 :肠复康合剂能抑制人结肠癌 HT- 2 9裸鼠移植瘤血管生成 ,其机制可能是降低瘤组织VEGF/ES比值。     

CIK细胞对裸鼠人胃癌移植瘤生长的抑制作用

目的: 探讨CIK细胞对裸鼠人胃癌移植瘤生长的抑制作用.方法: 用淋巴细胞分离液分离外周血单个核细胞, 给予多种细胞因子(rhIFN-γ、CD3mcAb、rhlL-2、rhlL-1), 诱导生成CIK细胞.培养人胃癌细胞株SGC-7901, 接种至40只裸鼠右腋皮下, 10 d后随机分2组, 每组20只, 分别为CIK组和对照组. 连续5 d在接种肿瘤细胞部位处给予CIK细胞和生理盐水注射治疗, 观察CIK细胞对胃癌移植瘤模型的抗肿瘤疗效.结果: CIK组胃癌肿块质量和生存期与对照组相比, 均具有显著性差异(1.21±0.34 g vs2.73±0.45 g, 65.8±6.2 d vs 44.3±4.8 d, 均P<0.01). 裸鼠体内实验表明, CIK细胞能够显著抑制胃癌细胞的生长, 其抑瘤率可达47.6%,明显高于对照组( P<0.01).结论: CIK细胞在裸鼠体内对人胃癌移植瘤有特异性抑瘤作用, 并可以延长裸鼠生存时间.     

罗格列酮对人结肠癌裸鼠移植瘤生长作用研究

目的观察过氧化物酶体增殖物激活受体的配体罗格列酮（ROZ）对人结肠癌细胞系HT-29裸鼠移植瘤的作用,探讨ROZ活化PPARγ,下调NFκB,从而诱导人结肠癌细胞凋亡的作用机制。方法体外培养人结肠癌HT-29细胞,建立人结肠癌细胞HT-29裸鼠移植瘤模型,20只荷瘤裸鼠随机分组进行实验。Western Blot法分析PPARγ、NF-κB、Bcl-2、bax蛋白表达的影响及PPARγ活化依赖性。结果 ROZ能抑制裸鼠移植瘤的生长。结论 ROZ通过上调PPARγ蛋白表达,下调NF-κB蛋白表达,抑制人结肠癌裸鼠移植瘤生长。     

Effects of TNP-470 on proliferation and apoptosis in human colon cancer xenografts in nude mice

AIM: To study the effect of TNP-470 on cell growth, proliferation and apoptosis in human colon cancer xenografts in nude mice. METHODS: Human colon cancer xenografts were transplanted into 20 nude mice. Mice were randomly divided into two groups. TNP-470 treated group received TNP-470(30 mg/kg, s.c) every other day and the control group received a sham injection of same volume saline solution. They were sacrificed after 4 weeks and their tumors were processed for histological examination. The expression of proliferating cell nuclear antigen (PCNA) in tumors was detected using immunohistochemical method with image analysis, and apoptosis in tumor cells was measured by TdT-mediated biotinyated-dUTP nick end labeling (TUNEL) staining. RESULTS: Comparing with controls, tumor growth was significantly inhibited in TNP-470 treated group, the inhibitory rate being 54.4 %. Expression of PCNA in tumors of TNP-470 treated group (PI 54.32+/-11.47) was significantly lower than that of control group (PI 88.54+/-12.36), P<0.01. Apoptosis index (AI) of TNP-470 treated group (18.95+/-1.71) was significantly higher than that of control group (7.26+/-1.44), P<0.001, typical morphological change of apoptosis in tumor cells was observed in TNP-470 treated group. CONCLUSION: Besides the anti-angiogenic effects, TNP-470 can inhibit tumor growth by inhibiting the proliferation and inducing apoptosis of tumor cells.     

Inhibitory effect of the angiogenesis inhibitor TNP-470 on tumor growth and metastasis in nude mice bearing human hepatocellular carcinoma

The antitumor and anti-metastatic effects of a potent angiogenesis inhibitor,O-(chloroacetyl-carbamoyl)fumagillol (TNP-470), was investigated in a highly metastatic model of human hepatocellular carcinoma—LCI-D20. Small pieces of LCI-D20 tumor tissue were implanted subcutaneously into the right axillary region of 24 nude mice; the mice were then randomized into two groups. To one group, TNP-470 30 mg/kg was given as a subcutaneous injection every other day from day 1 to day 15 and the mice were sacrificed on day 26. An antitumor effect of TNP-470 was clearly demonstrated by tumor weight (0.97±0.34 g compared to 2.04±0.34 g,P<0.001) and -Fetoprotein value (93±59 g/L compared to 769±282 g/L,P<0.001). There was also an anti-metastatic effect of TNP-470. Lung metastases developed in only 1 of 12 mice in the treated group, while they developed in 6 of mice of the control group. No severe side-effect of TNP-470 was found in this study. In vitro study revealed that the purified hepatoma cells were insensitive to TNP-470 (the 50% inhibitory concentration was 43 g/ml). These results suggest that the angiogenesis inhibitor TNP-470 has both strong antitumor and anti-metastatic effects on a human hepatocellular carcinoma model in nude mice.Abbreviations TNP-470 O-(chloroacetyl-carbamoyl) Fumagillol - MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide This work was partly supported by the CHina Medical Board of New York, grant 93-583, and a Leading Speciality grant of Shanghai Health Bureau     

Inhibitory effect of arsenic trioxide on peritoneal metastasis in nude mice with colon cancer

OBJECTIVE : To investigate the effects of As₂O₃ on the peritoneal metastasis of colon cancer in nude mice. METHODS : BALB/c‐nu/nu mice were injected intraperitoneally with Lovo cells and were allocated to five groups at random: control, epirubicin and varying concentrations of As₂O₃. The length of their survival period and rates of tumorigenesis and ascitic fluid formation were evaluated. RESULTS : Epirubicin and low doses of As₂O₃ clearly inhibited ascitic fluid formation and extended the survival period of tumor‐bearing nude mice. Moderate and large doses of As₂O₃ were able to eradicate cancer cells through apoptosis. CONCLUSIONS : As₂O₃ can induce apoptosis in intraperitoneal colon cancer cells, inhibit ascitic fluid formation and extend the survival periods of nude mice to varying degrees.     

烟曲霉醇联合5-氟尿嘧啶抑制大鼠结肠癌肝转移的实验研究

目的研究O-（氯乙酰-氨甲酰基）烟曲霉醇（TNP-470）联合5-Fu对大鼠结肠癌肝转移生长的抑制作用。方法建立大鼠结肠癌肝转移动物模型，设立不同的药物治疗组，观察肿瘤生长情况，测定转移灶中的微血管密度（MVD）及碱性生成纤维细胞生长因子（b-FGF）的表达。结果联合治疗组与TNP470组或5-Fu组比较，大鼠肝转移率显著下降（P〈0．05，P〈0．01）。与5-Fu组相比，转移灶中b-FGF表达和MVD计数均显著减少（P〈0．05，P〈0．01）。治疗前后大鼠体重无明显差异。结论TNP-470与5-Fu联合应用具有协同作用，可抑制结肠癌肝转移灶发生。     

胃康宁冲剂抑制胃癌生长与转移的实验研究

[目的]研究胃康宁含药血清在体内对胃癌细胞生长和转移的影响。[方法]将人胃癌MGC-803细胞注入祼小鼠腋前皮下,待生长成瘤后取出,将瘤块植入裸鼠胃壁内,建立胃癌原位移植瘤模型。设对照组、胃康宁高剂量组、胃康宁低剂量组,灌胃4周。检测各组原位肿瘤抑瘤率、肝及淋巴结转移情况。[结果]胃康宁高剂量组可明显抑制原位肿瘤的生长,对胃周淋巴结转移有明显抑制作用,对肝脏转移无明显抑制作用。[结论]胃康宁冲剂对胃癌的生长和转移有一定的抑制作用。     

Angiogenesis inhibitor TNP—470 suppresses growth of peritoneal disseminating foci of human colon cancer line Lovo

AIM：To study the effect of angiogenesis inhibitorTNP-470 on peritoneal dissemination of colon cancer in nude mice.METHODS:The MTT assay was used to evaluate the inhibitory effect of TNP-470on human colon cancer cell line Lovo.Lovo cells were injected into the peritoneal cavity of BABL/Cnu/nu mice and the models of peritoneal dissemination were developed,Thirty nude mice were randomly divided into control and TNP-470-treated group.In TNP-470-treated group,TNP-470 was injected subcutaneously every other day from day1until sacrifice or death(30mg&#183;kg^-1).The control group received a sham injection of the same volume saline solution.RESULTS:In vitro,TNP-470 inhibited the growth of Lovo cells,with its IC50at2.14&#215;10^2μg&#183;L^-1,In vivo,TNP-470 demonstrated growth inhibition of tumors.Mice body weight and abdominal circumferences were significantly different between TNP-470-treated group(24.5&#177;3.2g,7.0&#177;1.1cm)and control group(29.5&#177;2.1g,10.3&#177;1.5cm),P=0.005and P=0.001.The number of disseminated foci was significantly different between the control group(92.1&#177;20.6)and the TNP-470-treated group(40.3&#177;12.3),P&lt;0.001,The maximal size of foci was significantly smaller inTNP-470-treated group(3.3&#177;0.7mm)than that of control(7.3&#177;2.3mm),P=0.004,Mean survival time was significantly longer inTNP-470-treated group(98.00&#177;12.06d)than that in control group(41.86&#177;9.51d),P&lt;0.001.CONCLUSION:Angiogenesis inhibitorTNP-470 might be effective in treating peritoneal dissemination of colon cancer and improve the survival rate of nude mice.     

KDR单克隆抗体对荷人胃癌裸小鼠肿瘤生长的抑制作用

目的:研究血管内皮细胞生长因子(vascularendothelialgrowthfactor,VEGF)受体Ⅱ单克隆抗体(KDR-mAb)抑制荷人胃癌裸小鼠肿瘤生长的作用.方法:将人胃癌细胞(SGC-7901)通过背部皮下接种裸小鼠制备荷人胃癌小鼠肿瘤模型.待肿瘤生长至直径100-300mm3时将荷瘤小鼠随机分为3组,分别腹腔注射KDR-mAb、5-FU和生理盐水进行治疗.10d后剥离瘤组织,比较3组肿瘤组织的体积和质量、计算抑瘤率;通过Real-timePCR法和免疫组织化学法分别定量检测VEGF在各治疗组肿瘤组织中的表达.结果:KDR-mAb在体内能显著抑制实体瘤的生长,抑瘤率为36.3%,肿瘤大小和肿瘤质量与对照组比较差异显著(1889.94mm3±396.64mm3vs9398.34mm3±7413.96mm3,1.07g±0.58gvs1.68g±0.18g,均P<0.05);KDR-mAb能显著抑制肿瘤组织中VEGFmRNA和VEGF蛋白的表达(P<0.05).结论:作为VEGF受体抑制剂,KDR-mAb在荷人胃癌细胞小鼠肿瘤模型中具有明显的体内抗肿瘤活性,对于人胃癌的临床治疗具有一定的应用前景...     

Anti-tumor effect of angiogenesis inhibitor TNP-470 and 5-FU combined therapy on human gastric cancer xenograft

BACKGROUND/AIMS: TNP-470, an angiogenesis inhibitor, has already been used in combination with chemotherapy to enhance its antitumor activity. The mechanism of enhanced antitumor activity in combination therapy has not been clarified, however, and few studies have described the combined effect of TNP-470 and 5-fluorouracil (5-FU) on gastric cancer. The present study was conducted to investigate the effect of TNP-470 + 5-FU on gastric cancer cell line MKN-45 in vivo and in vitro. METHODOLOGY: MKN-45 cells were subcutaneously injected into mice that were divided into 4 groups: a control group, a 5-FU treated group, a TNP-470 treated group, and a 5-FU + TNP-470 treated group. After the inoculation, the volume of subcutaneous tumors was measured. Blood and lymphatic vessels were also analyzed for the assessment of lymphangiogenesis. RESULTS: Compared with 5-FU or TNP-470 alone, the combined effect of TNP-470 and 5-FU significantly inhibited and suppressed tumor growth in a synergistic fashion. The combined therapy significantly suppressed both angiogenesis and lymphagenesis. CONCLUSIONS: The study suggests that the combined therapy provides an enhanced antitumor effect on human gastric cancer. The enhanced antitumor activity is explained mainly by the stronger inhibition of angiogenesis.     

Augmented antitumour effects of combination therapy with TNP-470 and chemoimmunotherapy in mice

PURPOSE: To investigate antitumour efficacy of the combination of the antiangiogenic agent TNP-470 combined with chemoimmunotherapy in different tumour models in mice MATERIALS: B6D2F1 mice and BALB/c mice were inoculated in the footpad of the right hind limb with B16F10 melanoma cells or colon adenocarcinoma cells C-26, respectively. Subsequently, they received therapy consisting of TNP-470 and/or IL-12 and tumour growth was observed. In the melanoma model this therapy regimen was combined with cisplatin in a subtherapeutic dose. The antiangiogenic action of the tested agents was evaluated using tumour-induced angiogenesis assay in vivo. In order to analyse interactions between TNP-470 (or cisplatin) and IFN-gamma on tumour cells in vitro, the following methods were used: MTT assay, Western blot analysis, and flow cytometry analysis. RESULTS: Administration of the combined therapy with TNP-470 and IL-12 resulted in augmented antitumour activity in colon-26 and B16F10 melanoma models. Addition of cisplatin further enhanced efficacy of this combined therapy in the melanoma model. We showed that antitumour activity of this combined therapy is mediated by multiple mechanisms: not only is enhancement of the antiangiogenic activity mediated by TNP-470 and IL-12 but also by the synergistic cytostatic/cytotoxic action of IL-12-induced IFN-gamma and TNP-470 or cisplatin on tumour cells. The experiments revealed that TNP-470 together with IFN-gamma leads to the increased expression of p21 protein in cancer cells, which in turn may contribute to their cytostatic/cytotoxic action in vitro. CONCLUSION: Our experiments show a successful TNP-470-based combination therapy and suggest that the enhancement of the antitumour activity could be explained by a concomitant effect on both endothelial and tumour cell compartments.     

Angiogenesis inhibitor, TNP-470, prevents diet-induced and genetic obesity in mice

Adipose tissue growth has been proposed to involve recruitment of new blood vessels. Here, we test the hypothesis that delivery of an angiogenesis inhibitor in mice may prevent diet-induced obesity, the most common type of obesity in humans. We show that systemic administration of a selective angiogenesis inhibitor, TNP-470 (AGM-1470), prevents obesity in high caloric diet-fed wt mice as well as in genetically leptin-deficient ob/ob mice. Inhibition of obesity in mice by TNP-470 involves a reduction of vascularity in the adipose tissue. This therapeutic strategy appears to selectively affect the growth of adipose tissue as measured by the ratio between total fat and lean body mass. Interestingly, the treatment with TNP-470 results in decreased serum levels of low-density lipoprotein cholesterol. Furthermore, insulin levels are reduced, which indicates increased insulin sensitivity, suggesting that angiogenesis inhibitors may prevent the development of type II diabetes. Our findings suggest that similarly to growth and organogenesis in other tissues, adipose tissue growth is dependent on angiogenesis. Our observations may have conceptual implications for the prevention of obesity and related disorders.