槲皮素对人乳腺癌裸鼠移植瘤抑制作用的研究

目的 :研究槲皮素对人乳腺癌细胞株MCF 7裸鼠移植瘤的抑制作用及其对细胞增殖的影响。方法 :复制MCF 7裸鼠移植瘤动物模型 :2 4只移植成功的裸鼠分为对照组、槲皮素组、多柔比星组及联合用药组 4组 ,每组 6只。观察比较移植瘤的生长情况、移植瘤质量、光镜及电镜观察移植瘤结构的变化 ,Ki 67免疫组化分析对细胞增殖的影响。结果 :对照组移植瘤生长速度明显快于槲皮素组、多柔比星组及联合用药组。移植瘤质量对照组明显大于槲皮素组、多柔比星组及联合用药组 ,P <0 0 1。槲皮素组及联合用药组抑瘤率分别为 3 6 46%和 41 49%。Ki 67标记指数为对照组最高 ( 83 5 3 % ) ,槲皮素组次之( 66 95 % ) ,均高于联合用药组 ( 5 8 80 % ) ,P <0 0 1。结论 :槲皮素可抑制MCF 7裸鼠移植瘤生长及增殖 ,并可与多柔比星协同抑制移植瘤的增殖作用     

人乳腺癌裸鼠移植瘤MTD间歇期给予CTX节律化疗的实验观察

目的:研究最大耐受剂量(MTD)化疗间歇期给予节律低剂量CTX对人乳腺癌裸鼠移植瘤的作用.方法:建立人乳腺癌裸鼠移植瘤模型,随机分为小剂量节律(LDM)组、最大耐受剂量(MTD)组、最大耐受剂量联合节律(MTD+LDM)组和生理盐水对照组,治疗21 d为1个周期.测量瘤体积、裸鼠体质量及外周血白细胞计数.处死裸鼠后测量瘤质量,应用免疫组化方法检测瘤组织中血管内皮生长因子(VEGF)、微血管密度(MVD)及增殖细胞核抗原(PCNA)的表达.结果:各治疗组均抑制了肿瘤生长;联合组瘤质量及21d时肿瘤体积与其他各组相比均显著降低,P＜0.05.联合组抑瘤率为69.15％,与对照组及MTD组相比,联合组和LDM组肿瘤组织的VEGF水平及MVD值显著降低,P＜0.05;与LDM组和对照组相比,联合组和MTD组肿瘤组织PCNA表达水平均显著降低,P＜0.05.各治疗组裸鼠体质量及白细胞计数差异无统计学意义,P＞0.05.结论:最大耐受剂量间歇期给予节律低剂量CTX通过同时抑制肿瘤细胞增殖活性和肿瘤血管生成,在未增加毒副反应的前提下抑瘤效果更好,为可行有效的方案.     

三氧化二砷碘油乳化物对人MHCC-97裸鼠原位移植瘤模型抑瘤作用及药物释放的研究

目的:探讨三氧化二砷(As2O3)-碘油乳化物治疗裸鼠肝癌细胞株MHCC-97移植瘤模型的抑瘤作用及药代动力学情况.方法:将不同浓度的As2O3-碘油乳化物分别注入大白兔的腹腔内并检测外周血As2O3的浓度,绘制药物释放曲线,并将As2O3-碘油乳化物与人肝癌细胞株MHCC-97构建的肝脏型裸鼠肝细胞癌模型进行抑瘤作用.结果:3组大白兔腹腔内注入不同浓度梯度As2O3-碘油乳化物的实验组,与腹腔内单纯注入As2O3的对照组相比,30 min～1 h外周血中As2O3血药浓度偏低,P<0.05;2～3 h外周血中的血药浓度差异无统计学意义;在4～8 h血药浓度明显偏高,P<0.01;在3组实验组中,3种不同浓度梯度的As2O3-碘油乳化物在不同时间的血药浓度差异无统计学意义,P>0.05.采用As2O3-碘油乳化物对肝脏型裸鼠肝细胞癌模型的抑瘤作用与阴性对照组相比,差异有统计学意义,P<0.05;实验组与阳性对照组相比,平均瘤质量较阳性对照组有缩小,但无统计学意义,P>0.05.结论:As2O3-碘油乳化物在腹腔内释放缓慢,在外周血中能维持相对高浓度,并且维持时间长;As2O3-碘油乳化物对肝脏型裸鼠肝细胞癌模型的生长有抑制作用.     

抗乳腺癌人单克隆抗体用于裸鼠移植瘤显像

乳腺癌的发病率占恶性肿瘤的7～10％,是女性癌症致死的主要原因之一。在我国女性乳癌发病率约为23/10万人,仅次于子宫颈癌。以单克隆抗体为工具,探索乳腺癌的诊断和治疗问题,有重要临床价值。由于人体对鼠单克隆抗体产生明显排异反应,所以只有制备人单克隆抗体才能有效、安全地解决体内定位诊断和治疗的问题。为此,我们采用人鼠种间杂交骨髓瘤细胞SMH-D_(33)与人淋巴结B细胞融合,制备了人单克隆抗体CH·1(HMcAb-CM·1),进行实验研究,现报道如下。     

多肽p166结合顺铂对结肠癌裸鼠移植瘤抑制作用的观察

目的:观察计算机辅助药物设计合成的特异性多肽p166结合顺铂对人结肠癌裸鼠移植瘤的抑制作用及毒副反应,并探讨其作用机制.方法:建立人结肠癌细胞株HCT-116裸鼠皮下移植瘤模型,随机分为生理盐水组、顺铂组(1 mg/kg)和p166结合顺铂组(4 mg/kg,其中顺铂浓度为1 mg/kg).连续腹腔注射给药10 d,给药期间测瘤体体积计算抑瘤率,第11天处死裸鼠,采血测肝肾功能,剥离瘤体标本HE染色,采用免疫组化法检测凋亡因子Bcl-2、Bax、Cyt C和Caspase-3的表达.结果:p166结合顺铂组和顺铂组,抑瘤率分别为56.98%和41.96%.p166结合顺铂组血清BuN、Cr、ALT和AST水平均低于顺铂组,其中Cr、ALT和AST水平与顺铂组比较差异有统计学意义,P<0.05.免疫组化结果显示,与顺铂组比较,p166结合顺铂组Bcl-2蛋白表达减少,Bax、Cyt C和Caspase-3蛋白表达增强.结论:p166结合顺铂对HCT-116人结肠癌裸鼠移植瘤具有显著抑制作用,且能减少顺铂对裸鼠肝肾功能的影响.其机制可能与p166结合顺铂后能增加顺铂穿透肿瘤细胞胞膜的能力有关.     

胃泌素对人结肠癌裸鼠移植瘤细胞内cAMP含量的影响

目的 :探讨胃泌素对人结肠癌裸鼠移植瘤细胞内环磷酸腺苷 (cAMP)含量的影响。方法 :建立人结肠癌SW4 80 裸鼠移植瘤模型 ,实验动物随机分为 5肽胃泌素 (PG)组及对照组 (生理盐水 ) ,观察移植瘤的体积、重量及细胞内cAMP含量的变化。结果 :移植瘤的体积、重量、细胞内cAMP含量 ,PG组均显著高于对照组 (P <0 0 5 ,P <0 0 1)。结论 :胃泌素在体内使人结肠癌移植瘤细胞内cAMP含量增加 ,从而促进移植瘤的生长     

反义肝素酶cDNA对裸鼠人胰腺癌移植瘤生长和血管生成的抑制作用

目的研究反义肝素酶cDNA对裸鼠人胰腺癌SW1990细胞移植瘤生长和血管生成的抑制作用。方法将反义肝素酶cDNA转染成功的人胰腺癌SW1990细胞注射于裸鼠皮下建立移植瘤模型。观察移植瘤生长曲线、抑瘤率,以免疫组化方法检测移植瘤肝素酶表达和肿瘤微血管密度,比较反义组、空载体组和空白对照组的差异。结果反义组成瘤时间晚,瘤体大小较对照组小,t=4.11,P=0.00,抑瘤率在第6周时达64.2%。反义组、空载组和空白组移植瘤的免疫组化染色评分(IHS)分别为3.8±1.2、9.5±2.3和10.2±2.0,反义组与对照组比较,IHS明显降低,t=6.72,P=0.00。MVD计数分别为18.6±2.2,33.3±5.3和34.9±3.2条,与对照组比较,MVD明显降低t=10.28,P=0.00。结论反义肝素酶cDNA抑制裸鼠人胰腺癌移植瘤生长和血管生成,有希望成为一种有效的胰腺癌基因治疗方法。     

孕激素抗人乳腺癌的体内外研究

用人乳腺癌细胞株Ｂｃａｐ－３７、裸鼠移植性乳腺肿瘤模型及ＭＴＴ方法进行了孕激素醋酸甲地孕酮（ＭＡ）抗乳腺癌的体内外实验研究，并初步观察了高剂量ＭＡ治疗对乳腺癌病人的激素影响和疗效。结果显示：ＭＡ在体内外均有抗乳腺癌作用，呈现剂量－依赖和时间－依赖的特性，高剂量ＭＡ可导致乳腺癌病人激素水平的变化，部分病人获得疗效     

树突状细胞诱导抗裸鼠人肝癌移植瘤免疫

[目的]讨究树突状细胞（DC）体外诱导的抗肝癌细胞免疫能否预防裸鼠人肝癌移植癌发生及抑制课鼠人肝癌移植瘤生长。[方法]联合应用粒／巨噬细胞集落刺激因子（GM－CSF）及白介素－4（IL－4）直接从肝癌患者外周血中培养DC：以人肝癌细胞系HepG2肿瘤细胞的肿瘤抗原粗提物刺激DC；DC激活同源的T淋巴细胞产生细胞毒性T淋巴细胞（CYL）；以DC激活的CIL预防性接种于探鼠皮下观察其后接种的人肝癌细胞系HepG2移植瘤发生率；并观察该CTL对已接种裸鼠的人肝癌细胞系HepG2移植瘤生长的影响。[结果]DC诱导的工体外对HepG2肿瘤细胞具有高效而特异杀伤作用，在裸鼠体内不能预防探鼠人肝癌细胞系HepG2移植瘤发生，而且能抑制该移植瘤生长。[结论]经肝癌肿瘤相关抗原激活的肝癌患者外用血DC体外诱导的抗肿瘤细胞免疫在体内外均具有良好的抗肿瘤作用。提示DC作为一新概念上的抗肿瘤疫苗有可能在人肝癌的预防及治疗中发挥重要作用。     

常规化疗联合节拍化疗对乳腺癌裸鼠移植瘤的实验研究

背景与目的:乳腺癌治疗应从全局出发,将针对肿瘤细胞本身和肿瘤微环境相结合,而研究发现节拍化疗在抗血管生成等改善肿瘤微环境方面有明显优势.本文旨在观察环磷酰胺(cyclophosphamide,CTX)常规化疗联合节拍化疗对乳腺癌裸鼠移植瘤的抑瘤效应,探讨其对血管新生和细胞增殖、凋亡的影响.方法:建立乳腺癌裸鼠原位移植瘤模型,随机分成4组:节拍化疗(LDM)组、常规化疗(MTD)组、联合(LDM+MTD)组和0.9％NaCl溶液对照组,治疗期间观察裸鼠一般状况、隔日称重并测量皮下移植瘤体积,每周尾静脉采血白细胞计数(white blood cell counts,WBC).实验结束后处死小鼠取瘤称重,计算各组抑瘤率.免疫组织化学法检测移植瘤组织中微血管密度(microvessel density,MVD),以及血管内皮细胞生长因子(vascularendothelial growth factor,VEGF)、凝血酶敏感蛋白1(thrombospondin-1,TSP-1)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)表达,TUNEL检测肿瘤细胞凋亡.结果:CTX 3种化疗方案均可不同程度抑制移植瘤生长,其中LDM组和MTD组移植瘤生长曲线类似,LDM+MTD组移植瘤生长明显缓于其他各组;LDM组、MTD组和LDM+MTD组的抑瘤率分别为32.95％、41.57％和69.15％; LDM组和LDM+MTD组MVD、VEGF相对低表达、TSP-1相对高表达,而组间比较差异无统计学意义(P＞0.05),但与其余两组比较差异有统计学意义(P＜0.05).MTD组和LDM+MTD组PCNA相对低表达,与其余两组比较差异有统计学意义(P＜0.05),LDM+MTD组凋亡指数(apoptosis index,AI)明显高于其他各组,差异有统计学意义(P＜0.05).结论:CTX常规化疗联合节拍化疗兼有抗血管生成、抑制细胞增殖、促进肿瘤细胞凋亡作用,抑瘤作用较单一传统化疗和节拍化疗更为明显,不良反应不明显.     

Radiotherapy alternated with chemotherapy and immunotherapy on nude mice bearing human hepatocellular carcinoma

A comparative study of multiple modalities of multiple fractions of external radiation per day (MFD), routine radiation (RD), cisplatin (CDDP), mixed bacterial vaccine (MBV), MFD + CDDP. MFD + MBV, MFD + CDDP + MBV on nude mice bearing human hepatocellular carcinoma (HCC) were investigated. The tumor size and serum alpha fetoprotein (AFP) level were measured. The results showed that the tumor inhibition rates were 97%, 94%, 79%, 2% for the MFD, RD, CDDP, and MBV groups, respectively. No tumor disappeared (complete regression) in single modality groups. When MFD was used in combination with CDDP and/or MBV, the tumor cure rates were remarkably increased. The tumor inhibition rate was 99% in the MFD + MBV group, in which two tumors disappeared, 99% in the MFD + CDDP group as well, in which five tumors disappeared. The greatest effectiveness was observed in the MFD + MBV + CDDP group with tumor inhibition rate of 100%, in which eight tumors disappeared. In all these groups, the AFP level was decreased as the tumor size reduced. Preliminary clinical results were satisfactory, the decrease of the serum AFP level and shrinkage of tumor were observed in 88% (7/8) and 91% (10/11) of patients, respectively.     

Experimental chemotherapy of human testicular carcinoma transplanted in nude mice

The testicular carcinoma serially transplanted in nude mice with BALB/c genetic background was used for experimental chemotherapy. A stable growth and a high production of alpha-fetoprotein (AFP) were observed in this tumor line. The effect and side effect of Cis-platinum (CDDP) and other anticancer agents on this tumor line in nude mice were studied by the chemotherapy with single administration of CDDP 2 mg/kg, 4 mg/kg, 6 mg/kg and 8 mg/kg, and the combination chemotherapy with CDDP, Vinblastine (VBL) 0.1 mg/kg, Bleomycin (BLM) 0.5 mg/kg and Cyclophosphamide (CTX) 2 mg/kg. The body weight of the tumor bearing nude mice, the tumor size (length X breadth) and serum AFP level were measured every week up to 10 weeks after inoculation of the tumor mass. Six weeks after administration of these anticancer agents, the tumor mass was removed out and examined histologicaly. The effects of CDDP and other anticancer agents were observed as inhibition of the tumor growth and regression of the tumor mass. In the groups treated by the combination chemotherapy with either CDDP + VBL + BLM or CDDP + VBL + CTX, the most remarkable inhibition and regression were observed. The AFP levels were remarkably decreased in contrast with those of the control group. The changes of serum AFP levels were reflected in the tumor growth. The serum AFP levels fell down to normal level, however, the tumor mass was clearly recognized. The tumor tissue was damaged histologicaly by the single administration of CDDP. The most remarkable change was shown in the group treated by the combination chemotherapy CDDP 4mg/kg + VBL + BLM. The tumor cells were arranged one or two layers like the epithelium. This histological findings suggested that the malignant tumor could be differentiated to benign tumor. The side effect of CDDP and other anticancer agents was observed as a loss of weight. All of mice treated by the single administration at a dose of CDDP 6 mg/kg and 8 mg/kg died of the side effect of CDDP.     

Radioimmunotherapy combined with chemotherapy and immunotherapy for nude mice bearing human hepatocellular carcinoma

A comparative study of multiple modalities, radioimmunotherapy combined with cisplatin and MBV was made. The tumor size and macrophage activity (acid phosphatase) were measured after treatment. The results showed that the tumor inhibition rates were 48, 55, 74, 76, 79% in radioimmunotherapy, cisplatin, radioimmunotherapy + MBV, radioimmunotherapy + cisplatin and radioimmunotherapy + MBV + cisplatin groups, respectively. Radioimmunotherapy was effective in controlling tumor growth, especially in sequential treatment by two injections. Both cisplatin and MBV could increase therapeutic effect of radioimmunotherapy. Therefore, combination of the three modalities is the best choice for tumor growth control. The effectiveness of MBV may be related to the increase of macrophage activity. Preliminary clinical results were satisfactory. Decline in serum AFP level and shrinkage of tumor were observed in 80% (12/15) and 65% (13/20) of the patients. It is suggested that combination of multiple treatment modalities may provide an important approach to treat moderately advanced liver cancer.     

Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice

There are few reports describing experimental models of the growth and metastasis of human breast carcinomas. This article discusses the tumorigenic and metastatic properties of two estrogen receptor-negative breast carcinomas injected into nude mice. Tumor growth in the mammary fatpad (m.f.p.) and the subcutis was compared in female nude mice. The injection of 10(5) viable cells of two human breast carcinoma cell lines (MDA-MB-231 and MDA-MB-435) gave a 100% tumor take rate in the m.f.p., whereas only 40% of the s.c. injections produced tumors and these occurred several weeks after the appearance of the m.f.p. tumors. Thus, the m.f.p. of nude mice is a favorable site for the growth of human breast carcinomas. MDA-MB-435 tumors produced distant metastases in 80% to 100% of recipients. The most common sites for metastasis were the lymph nodes and lungs, with a lower incidence of metastases in muscle (chest wall and thigh), heart, and brain. New variant cell lines were isolated from metastases in the lungs, brain, and heart. All the cell lines were tumorigenic in the m.f.p., and the lung- and heart-derived metastasis lines produced slightly more lung metastases than the original cell line. However, the brain metastasis variant produced significantly fewer lung metastases. Intravenous inoculation of the spontaneous metastasis-derived cell lines produced few lung colonies. Only cell variants isolated from experimental lung metastases showed enhanced lung colonization potential when reinjected i.v. Our results suggest that the estrogen receptor-negative MDA-MB-435 cell line injected in the m.f.p. of nude mice could be a valuable tool for analysis of the cellular and molecular basis of the metastasis of advanced breast cancer.     

Antitumor effect of trimelamol against human breast carcinoma xenografts in nude mice

The antitumor effect of N-2, N-4, N-6-trihydroxymethyl-N-2, N-4, N-6-trimethylmelamine (trimelamol), a synthetic analogue of hexamethylmelamine, was investigated using human breast carcinoma xenografts in nude mice. Four tumor models, T-61, Br-10, R-27 and MCF-7 were estrogen receptor (ER)-positive and their growth was estradiol-dependent. The MX-1 model was ER-negative and grew estradiol-independently. Sixty mg of trimelamol per kg dissolved in 5% dimethylsulphoxide (DMSO) with 5% glucose was administered intraperitoneally for 5 days weekly for three weeks. Trimelamol showed potent antitumor activity on T-61 and MX-1 in a dose-responsive manner with a marginal effect on Br-10, whilst R-27 and MCF-7 were insensitive to this agent. This antitumor spectrum on human breast carcinoma xenografts was similar to that of hexamethylmelamine previously reported using the same xenograft models. Trimelamol is water-soluble and does not require metabolic activation which is needed for hexamethylmelamine. These advantages allow the paraenteral administration of trimelamol, and warrant the further investigation of this drug for breast carcinomas.     

Hyperthermia enhances mitoxantrone cytotoxicity on human breast carcinoma and sarcoma xenografts in nude mice.

In this preclinical in vivo study, we measured antitumor response, local side effects and systemic toxicity of locally applied water-bath hyperthermia given alone or simultaneously with mitoxantrone (3 mg/kg b.w. i.v.; LD 10) on a human derived breast carcinoma (MX 1) or a human sarcoma (S 117) transplanted to female athymic (nude) mice. A single hyperthermia treatment at a tumor temperature up to 43 degrees C for 1 hr caused in both tumor lines only minor tumor regressions and transient tumor growth delay. However, the antitumor effect of mitoxantrone was significantly enhanced by local hyperthermia at 42 degrees C and particularly at 43 degrees C. In both tumor lines complete tumor regressions were achieved only if mitoxantrone was combined with hyperthermia. Undesired side effects and drug toxicity were not enhanced by hyperthermia. According to in vitro data and the results of the present in vivo study mitoxantrone seems to be a good candidate for clinical trials in combination with locoregional hyperthermia.     

N-acetylglucosaminyltransferase V activity in metastatic models of human hepatocellular carcinoma in nude mice

N-linked beta 1-6 branched oligosaccharides may contribute directly to the malignant phenotype including metastatic potential of tumour cells. Increased beta 1-6 branching was associated with an increased level of N-acetylglucosaminyltransferase V (GnT V). In this report, the tissues from two metastatic models of human hepatocellular carcinoma (HCC) in nude mice were obtained. GnT V activity and mRNA level were determined. Results showed that GnT V activity in highly metastatic LCI-D20 models (Liver Cancer Institute, passage time: 20 days) (413.1+/-86.4U) was much higher than that in low metastatic LCI-D35 model (passage time 35 days) (155.3+/-31.9U). Northern blot showed that the mRNA level of GnT V in two models had no change. During the selection of a highly metastatic LCI-D20 model, GnT V activity increased from 301.6+/-57.3U to 413.1+/-86.4U while the highly metastatic LCI-D20 model acquired higher metastatic ability after selection. When highly metastatic LCI-D20 model tissues were implanted subcutaneously (s.c.), the GnT V activity decreased dramatically from 413.1+/-86.4U to 94.9U. This is the first report that GnT V activity increased in HCC during metastasis in vivo.