Modulation of bone microenvironment with zoledronate enhances the therapeutic effects of STI571 and paclitaxel against experimental bone metastasis of human prostate cancer

Prostate cancer cells metastasize to the bone where their interaction with osteoclasts and osteoblasts can lead to alterations in the structure of the bone. We determined whether the systemic administration of the bisphosphonate, zoledronate, could prevent bone lysis and halt the proliferation of human prostate cancer cells injected into the tibia of nude mice. Zoledronate did not affect the in vitro proliferation of human prostate cancer PC-3MM2 cells. The in vivo administration of zoledronate produced significant bone preservation but did not inhibit the progressive growth of PC-3MM2 cells. The systemic administration of STI571 (imatinib mesylate, Gleevec), an inhibitor of phosphorylation of the platelet-derived growth factor receptor, in combination with paclitaxel, produced apoptosis of tumor cells and bone- and tumor-associated endothelial cells. The systemic administration of zoledronate with STI571 and paclitaxel produced a significant preservation of bone structure, a decrease in tumor incidence and weight, and a decrease in incidence of lymph node metastasis. This therapeutic activity was correlated with inhibition of osteoclast function, inhibition of tumor cell proliferation, and induction of apoptosis in tumor-associated endothelial cells and tumor cells. Cancer is a heterogeneous disease that requires multimodality therapy. The present data recommend the combination of a bisphosphonate agent with protein tyrosine kinase inhibitor and an anticycling drug for the treatment of prostate cancer bone metastasis.     

去甲斑蝥素对NCI-H446骨转移裸鼠模型的干预作用

目的探讨去甲斑蝥素对肺癌骨转移的抑制作用。方法通过贴骨接种NCI．H446肺癌细胞制作骨转移裸鼠模型,将小鼠分为对照组、低剂量药物组、高剂量药物组等3组,评价各组药物干预骨转移的情况。观察3组小鼠肿瘤生长的变化。对各组小鼠进行x线比较及解剖、病理学观察。分别检测早期、晚期骨转移阶段各组血清PICP、PINP及BSP水平,并进行统计学分析。结果与药物组比较,对照组肿瘤生长快,肿瘤质量、体积大,实验结束时体重增长总量显著低于药物组;对照组PICP、PINP水平均明显升高;低剂量药物组PICP水平明显升高、PINP水平略有增高;高剂量药物组放射学检查显示晚期肿瘤性骨损伤组血清PICP、PINP水平升高均不明显;各组血清BSP水平均升高。结论去甲斑蝥素对肺癌骨转移具有抑制作用,血清PICP、PINP水平对小细胞肺癌骨转移具有早期诊断及疗效监测作用。     

Ad-mda-7联合多柔比星对人肝癌裸鼠移植瘤的协同抑制作用

目的:将重组腺病毒介导的黑素瘤分化相关基因-7(melanoma differentiation-associated gene-7,mda-7)/IL-24与多柔比星(adriamycin,ADM)联合治疗人肝癌细胞裸鼠移植瘤,探索基因治疗和化疗相结合治疗肝癌的新方法.方法:构建携带人mda-7/IL24基因的重组腺病毒载体(Ad-mda-7),单独使用该载体或ADM以及两者联合使用对人肝癌细胞裸鼠移植瘤进-行治疗,观察抗肿瘤效果.用免疫组织化学方法检测各组肿瘤组织中基质金属蛋白酶-2(matrix metalloproteinase,MMP-2)与转化生长因子(transforming growth factor,TGF)-β1的表达.结果:成功构建重组腺病毒并实现体内表达,Ad-mda-7 ADM联合治疗组的裸鼠平均生存时间为(83.8±4.82)d,与其他3组相比生存期明显延长(P<0.01);Ad-mda-7 ADM组裸鼠移植瘤平均大小与单独使用ADM或Ad-mda-7相比明显缩小(P<0.01).Ad-mda-7能显著降低TGF-β1和MMP-2的表达,明显优于ADM组(P<0.01).结论:重组腺病毒介导mda-7/IL-24联合多柔比星对裸鼠人肝癌转移模型有明显的抗肿瘤作用及协同效应,其机制可能与抗肿瘤侵袭转移有关.     

辛伐他汀在裸鼠模型中对乳腺癌骨转移的影响

目的 观察辛伐他汀在裸鼠模型中对乳腺癌骨转移的作用.方法采用完全随机分组方法 将60只裸鼠分为3组,每组20只,裸鼠左心腔注射乳腺癌骨转移细胞株(MDA-MB-231),7d后,分别皮下注射辛伐他汀、生理盐水及无任何处理(2次/周,19 d).应用图像分析软件评估骨转移瘤的面积.随后处死裸鼠,用放射免疫法检测骨转移癌髓腔内甲状旁腺素相关蛋白(PTHrP)浓度;应用骨密度检测软件进行组织形态学分析,计数骨转移灶每毫米癌组织与临近骨小梁之间破骨细胞的数量.计量资料比较采用方差分析,P＜0.05为差异有统计学意义.结果 与生理盐水组和无处理组相比,注射辛伐他汀的裸鼠骨转移癌面积明显减小(0.51±0.18 mm2 vs 2.13±1.24 mm2 vs 2.29±1.22 mm2;F=15.600,P=0.002; F=15.673,P=0.001),骨转移癌周围髓腔内PTHrP浓度明显降低(0.98±0.20 pmol/L vs 2.11±0.31 pmol/L vs 1.99±0.29 pmol/L;F=61.469,P＜0.001;F=58.274,P＜0.001),并且其转移灶破骨细胞的数量明显减少(4.00±1.73个/mm vs 11.40 ±4.93个/mm vs 10.91±3.87个/mm;F=17.820,P=0.001,F=17.184,P=0.002).结论 辛伐他汀能够降低乳腺癌细胞PTHrP的分泌,从而抑制乳腺癌细胞在骨内生长及其对骨质的破坏.     

Weekly administration of paclitaxel induces long-term aneugenicity in nude mice

We investigated the potential in vivo aneugenic effects associated with paclitaxel treatment. For this purpose, we treated female nude mice with paclitaxel using doses equivalent to those used in weekly schedules at the clinical level (three cycles of 30 mg/kg/week for three consecutive weeks followed by one resting week). We then evaluated the frequencies of micronucleated erythrocytes (MNE) in peripheral blood using the acridine orange micronucleus assay. The frequency of MNE was evaluated after 24 h and 168 h of administration of the last dose of each paclitaxel cycle (STA mice group) as well as after one year of the first dose of treatment (LTA mice group). We also analyzed the cytology of peripheral blood and bone marrows obtained from these mice at each time period. In the STA mice group, three cycles of paclitaxel induced a 2.4-fold increase in MNE frequencies compared to the control group (p < 0.01). This effect was observed after 24 h of the last dose of each chemotherapy cycle and persisted at least for 168 h. In the LTA mice group, paclitaxel-treated mice presented a 1.8-fold increase in the MNE frequency (p = 0.01) indicating that paclitaxel-induced MNE increase lasted for at least one year. Although the appearance of micronuclei in erythrocytes and granulocytes in peripheral blood and bone marrow cytological smears, there was no evidence of myeloproliferative disease. The present data therefore indicate an aneugenic potential of paclitaxel for humans, which should be considered in the risk-benefit analysis of its increasing clinical use.     

Hyaluronectin modulation of lung metastasis in nude mice

Hyaluronectin (HN) is a glycoprotein with a high affinity to hyaluronic acid (HA) and known to be a component of the extracellular matrix of tumours. Clinical studies have shown that a low level of HN correlates to tumours with poor prognosis, whereas a high level of HN correlates to tumours with good prognosis. We previously demonstrated in vitro that hyaluronidase activity, which promotes tumour progression and metastatic spread by degradation of HA into angiogenic oligosaccharides, was inhibited or promoted by HN, according to the level of HN-expression. This raises the question of the role played by HN in cancer, and particularly if high and low levels of HN-expression could trigger opposite effects on tumour growth and/or metastatic spread. To address this issue, we used a model of spontaneous lung fluorescent metastases that we characterised previously. We stably transfected the human HN cDNA into fluorescent H460M^GFP cells and selected two clones characterised by different levels of HN-expression: HN110 and HN704, with a high and a low level of HN-expression, respectively. In vitro, we demonstrated that HN704 cell migration was significantly increased. Inoculation of clones to nude mice had no significant effect on tumour growth, but clearly revealed opposite effects on metastatic spread: HN110 significantly decreased the number of fluorescent metastases whereas HN704 significantly increased it. We also analysed HN, HA and hyaluronidase contents in sera and tumours. These results demonstrate that HN can play a role as either a suppressor or promoter of metastatic spread.     

Simultaneous blockade of platelet-derived growth factor-receptor and epidermal growth factor-receptor signaling and systemic administration of paclitaxel as therapy for human prostate cancer metastasis in bone of nude mice

Once prostate cancer metastasizes to bone, conventional chemotherapy is largely ineffective. We hypothesized that inhibition of phosphorylation of the epidermal growth factor receptor (EGF-R) and platelet-derived growth factor receptor (PDGF-R) expressed on tumor cells and tumor-associated endothelial cells, which is associated with tumor progression, in combination with paclitaxel would inhibit experimental prostate cancer bone metastasis and preserve bone structure. We tested this hypothesis in nude mice, using human PC-3MM2 prostate cancer cells. PC-3MM2 cells growing adjacent to bone tissue and endothelial cells within these lesions expressed phosphorylated EGF-R and PDGF-R alpha and -beta on their surfaces. The percentage of positive endothelial cells and the intensity of receptor expression directly correlated with proximity to bone tissue. Oral administration of PKI166 inhibited the phosphorylation of EGF-R but not PDGF-R, whereas oral administration of STI571 inhibited the phosphorylation of PDGF-R but not EGF-R. Combination therapy using oral PKI166 and STI571 with i.p. injections of paclitaxel induced a high level of apoptosis in tumor vascular endothelial cells and tumor cells in parallel with inhibition of tumor growth in the bone, preservation of bone structure, and reduction of lymph node metastasis. Collectively, these data demonstrate that blockade of phosphorylation of EGF-R and PDGF-R coupled with administration of paclitaxel significantly suppresses experimental human prostate cancer bone metastasis.     

Synergistic inhibitory effect of wogonin and low-dose paclitaxel on gastric cancer cells and tumor xenografts

Objective：To investigate the synergistic inhibitory effects of wogonin （WOG） and chemotherapeutic drugs on growth of gastric cancer cells and tumor xenografts.Methods：The IC50 values of WOG,cisplatin （CDDP） and paclitaxel （PTX） in four gastric cancer cell lines were determined by MTS assay.Hoechst staining and the median effect method of Chou-Talalay were used to assess the apoptosis of cells and the interaction of two drugs,respectively.BGC-823-derived xenografts in nude mice were established to investigate the effects of WOG combined with chemotherapeutic drugs in vivo.Results：WOG,CDDP and PTX inhibited the growth of BGC-823,MGC-803,MKN-45 and HGC-27 gastric cancer cells in a dose-dependent manner.WOG combined with CDDP or PTX synergistically inhibited the growth of all gastric cancer cell lines in vitro.In BGC-823,MGC-803,HGC-27 and MKN-45 cell lines,synergisms between WOG and PTX were shown when the fraction affected （Fa） values were ＜0.45,＜0.90,＜0.85 and ＜0.60.While WOG and CDDP had a synergistic inhibitory.effect when the Fa values were ＞0,＞0,＞0.65 and ＞0.10.From the results of in vivo experiments using tumor xenografts,WOG and low-dose PTX showed better efficacy than either drug alone.The inhibitory percentages of tumor weight were 61.58％,20.29％,and 22.28％ for the combination,WOG-alone,and low-dose PTX-alone groups,respectively.Notably,WOG combined with CDDP displayed very high toxicity.Conclusions：A synergistic inhibitory effect on growth was observed when WOG was combined with low-dose PTX in gastric cancer cells and tumor xenografts.These findings provide evidence for the design of a clinical trial to test the combination of WOG with low-dose PTX in human gastric cancer.     

Effects of bisphosphonate on experimental jaw metastasis model in nude mice

The mechanism of osteolysis associated with metastatic cancer of the jaws is essentially osteoclast-mediated. Therefore, it is likely that potent osteoclastic bone resorption inhibitors such as bisphosphonates would be efficacious for the treatment of jaw metastasis. We examined the effects of a third generation bisphosphonate, YM175, in a nude mice jaw metastasis model with intracardiac injection of a human breast cancer cell line, MDA-MB-231. The metastatic lesions in untreated mice were radiographically observed at the body and angle of the mandible. Histology of the mandible of untreated mice revealed that most of the bone marrow cavities had been occupied by the metastatic tumor with active osteoclasts along the trabecular bone. The experimental group showed that YM175 markedly reduced the size of tumor and the number of osteoclasts. These results suggest that YM175 may suppress metastasis formation and tumor growth in jaw through inhibition of osteoclastic bone resorption.     

Tumor-targeting Salmonella typhimurium A1-R prevents experimental human breast cancer bone metastasis in nude mice

Bone metastasis is a lethal and morbid late stage of breast cancer that is currently treatment resistant. More effective mouse models and treatment are necessary. High bone-metastatic variants of human breast cancer cells were selected in nude mice by cardiac injection. After cardiac injection of a high bone-metastatic variant of breast cancer, all untreated mice had bone metastases compared to only 20% with parental cells. Treatment with tumor-targeting Salmonella typhimurium A1-R completely prevented the appearance of bone metastasis of the high metastatic variant in nude mice (P < 0.001). After injection of the highly bone-metastatic breast cancer variant to the tibia of nude mice, S. typhimurium A1-R treatment significantly reduced tumor growth in the bone (P < 0.001). These data indicated that S. typhimurium A1-R is useful to prevent and inhibit breast cancer bone metastasis and should be of future clinical use for breast cancer in the adjuvant setting.     

紫杉醇联合奈达铂对HeLa细胞移植瘤生长的影响

背景与目的:紫杉醇(TXL)联合顺铂(DDP)已成为治疗宫颈癌的常用化疗方案,但DDP毒性反应大.新一代铂类制剂奈达铂(NDP)对宫颈癌抗瘤活性高,毒性低.本实验通过与TXL DDP方案进行比较,研究TXL NDP对HeLa细胞裸鼠皮下移植瘤的抑制作用以及毒副反应.方法:以HeLa细胞裸鼠皮下移植瘤为研究对象,用抽签法将裸鼠随机分为6组(对照组,TXL组,NDP组,DDP组,TXL NDP组,TXL DDP组),观察肿瘤体积及体重变化.单药组评价用对照组(C)和治疗组(T)肿瘤体积比率T/C评价生长抑制,应用合用率评价联合用药,体重减轻超过20%为达到毒性范围.结果:TXL NDP与TXL或NDP相比,对HeLa细胞生长抑制作用明显[(172±60)mm3 vs(304±55)mm3,P＜0.001;(172±60)mm3 vs(356±67)mm3,P＜0.001],TXL与NDP合用呈协同作用(CR=0.76),而且最大体重减轻率为8.3%,未达到毒性范围.TXL DDP抗癌活性与TXL NDP基本等同[(198±77)mm3 vs (172±60)mm3,P＞0.05],但毒性反应增大,有4只裸鼠死亡.结论:TXL NDP对HeLa细胞抗癌活性与TXL DDP相似,但毒性反应小,有临床应用前景.     

Studies of human breast cancer metastasis using nude mice

Athymic nude mice have been used in recent years to study the biology of human tumors and to assess therapeutic responses in vivo rather than just in vitro. Some human tumors metastasize in nude mice, providing model systems for analyzing various aspects of the metastatic phenotype of human neoplasms. For breast carcinomas, however, the tumor-take rate of surgical specimens is low, and only a limited number of cell lines proliferate in nude mice. The site of injection of the breast carcinoma cells is important; tumors grow at a lower inoculum dose and with shorter latent intervals after implantation in the mammary fatpad of nude mice than after injection in the subcutis. One breast carcinoma cell line, MDA-MB-435, metastasizes from mammary fatpad tumors to lymph nodes, lungs, and other visceral organs. In contrast, two other cell lines show lower metastatic ability. Intravenous injection and injection of tumor cells into the internal carotid artery of nude mice produces lung and brain metastases, respectively, thus simulating the arrest and organ colonizing steps of the metastatic cascade. These different techniques demonstrate the potential of experimental studies of human breast cancer growth and metastasis using nude mice.     

Synergistic therapy of human ovarian carcinoma implanted orthotopically in nude mice by optimal biological dose of pegylated interferon alpha combined with paclitaxel.

The purpose of this study was to optimize the antitumor andantiangiogenic activities of pegylated IFN-alpha (PEG-IFN-alpha)alone or in combination with paclitaxel against SKOV3ip1 human ovarian cancer cells growing orthotopically in female nude mice. Seven days after the i.p. implantation of tumor cells, groups of mice (n = 10) were injected s.c. once per week (for 4 weeks) with different doses of PEG-IFN-alpha (3,500, 7,000, 35,000, and 350,000 units). PEG-IFN-alpha at 7,000 units significantly decreased tumor incidence and volume. At doses exceeding 7,000 units, PEG-IFN-alpha was less efficacious. In another set of studies conducted 7 days after the i.p. implantation of SKOV3ip1 cells, groups of mice (n = 10) received (once per week for 4 weeks) either s.c. administrations of PEG-IFN-alpha (7,000 units), i.p. injections of paclitaxel (100 microg/wk), or a combination of PEG-IFN-alpha and paclitaxel. The mice were killed 7 days after the last treatment, and tumor burden was assessed. Administration of PEG-IFN-alpha at the optimal biological dose (7,000 units) in combination with paclitaxel significantly decreased angiogenesis and progressive growth of human ovarian carcinoma cells in a synergistic fashion. The combination therapy produced the most significant inhibition in expression of the proangiogenic molecules basic fibroblast growth factor and matrix metalloproteinase-9. Decreased microvessel density, decreased proliferating cell nuclear antigen staining, and increased endothelial cell apoptosis also correlated with therapeutic success. Collectively, the data suggest that combining the optimal biological dose of PEG-IFN-alpha with paclitaxel may provide a novel and effective approach to the treatment of human ovarian carcinoma.     

Synergistic antitumor activity of combination chemotherapy with mitomycin C and cisplatin against human gastric cancer xenografts in nude mice

A new combined cancer chemotherapy regimen of mitomycin C (MMC) and cisplatin (DDP) showed synergistic antitumor activity against human gastric cancer xenografts St-40 and SC-1-NU in BALB/c nu/nu mice. The drugs were administered intraperitoneally at doses of 2 or 4 mg/kg for MMC and 3 or 6 mg/kg for DDP, respectively. To clarify the schedule-dependent antitumor activity of MMC and DDP against St-40 and SC-1-NU, different sequential therapies were conducted. Simultaneous administration of these agents showed the highest antitumor activity against SC1-NU among the three regimens used, whereas the sequence of MMC followed by DDP showed higher antitumor activity than the reverse sequence against St-40. The intratumoral concentration of platinum was significantly increased in St-40 treated with the sequence MMC to DDP, in comparison with the sequence DDP to MMC. The maximum tolerated dose (MTD) of this combination was 4 mg MMC plus 6 mg DDP per kg in all the combinations, and these MTDs were 2/3 of the corresponding values for their single use. Since this combination increased the antitumor activity of each single agent without any increase in their toxicity, it would appear to be useful clinically. © 1994 Wiley-Liss, Inc.     

Evaluation of paclitaxel against human adenocarcinoma of the uterine cervix in nude mice

Background The purpose of this study was to describe a new multidrug regimen for treatment of adenocarcinoma of the uterine cervix. Methods The cytotoxicity of paclitaxel against 3 cell lines (TCO, TCM, and TCY) of human adenocarcinoma of the uterine cervix transplanted into nude mice was studied. Monotherapy with paclitaxel, cisplatin, or mitomycin C, and combination therapy with paclitaxel+cisplatin, paclitaxel+mitomycin C, and cisplatin+mitomycin C, were compared with a control group injected with saline. Results The relative tumor growth rates (%) 4 weeks after the initial injections (after 3 weekly injections) for the TCO, TCM, and TCY tumor transplants were calculated. Paclitaxel mono- and combination-therapies were most effective against TCO (paclitaxel, 18.8%; cisplatin, 106.7%; mitomycin C, 53.9%; paclitaxel+cisplatin, 35.2%; paclitaxel+mitomycin C, 36.6%; and cisplatin+mitomycin C, 54.5%. Paclitaxel monotherapy was not effective against TCM, but combination therapies including paclitaxel were effective against TCM (paclitaxel, 53.8%; cisplatin, 53.1%; mitomycin C, 48.6%; paclitaxel+ cisplatin, 23.3%; paclitaxel+mitomycin C, 20.0%; and cisplatin+mitomycin C, 38.4%). Lastly, all dosing regimens, except for paclitaxel monotherapy, were effective against TCY (paclitaxel, 64.5%; cisplatin, 4.1%; mitomycin C, 2.9%; paclitaxel+cisplatin, 4.5%; paclitaxel+mitomycin C, 5.1%; and cisplatin+ mitomycin C, 5.9%). Conclusion It has been suggested that the addition of paclitaxel might make chemotherapeutic regimens more effective for tumors resistant to cisplatin-or mitomycin C, because paclitaxel tumor suppression acts through a different pathway. We concluded that paclitaxel combination therapy is effective against adenocarcinoma of the uterine cervix.     

原发瘤切除对荷人骨肉瘤裸鼠血管生成和肺转移的影响

目的 研究原发瘤切除对骨肉瘤血管生成和肺转移的影响,并探讨其可能的机制及临床意义.方法 用细胞悬液法构建倚人骨肉瘤裸鼠模型,分为实验组(切除原发瘤)、截肢对照组(不切除原发瘤的截肢手术)和空白对照组.用酶联免疫吸附试验(ELISA)法,检测手术前后裸鼠血清中血管内皮生长因子(VEGF)和内皮抑素的表达水平.用HiCN法检测裸鼠体内Matrigel胶体中血红蛋白浓度.应用膨胀压片计数法和组织切片苏木素-伊红对比染色法,观察肿瘤肺部转移灶的情况.结果 手术后实验组血清中的VEGF和内皮抑素浓度较空白对照组和截肢对照组有明显的下降,且内皮抑素下降幅度更大[VEGF分别为(71.43±9.15)pg/ml、(115.81±4.38)pg/ml和(111.68±12.26)ps/ml(P<0.01);内皮抑素分别为(40.77±5.41)ng/ml、(123.18±5.94)ng/ml和(128.06±4.52)ng/ml(P<0.01)];实验组、空白对照组和截肢对照组Matrigel小体中血红蛋白浓度分别为(36.55±2.35)g/L、(16.84±1.15)g/L和(16.29±1.10)g/L(P<0.01).实验组、空白对照组和截肢对照组的肺转移率分别为80.0%、40.0%和35.0%,差异有统计学意义(P<0.05).结论 原发瘤切除可使荷人骨肉瘤裸鼠体内的VEGF和内皮抑素比例失衡,有利于肿瘤血管生成和肺转移,故原发瘤切除后及时抑制肿瘤血管生成,对降低骨肉瘤肺转移有重要意义.     

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.