赛来昔布对人骨肉瘤类肿瘤干细胞移植瘤微血管生成的影响

[目的] 探讨cox-2抑制剂赛来昔布(Celecoxib)对骨肉瘤类肿瘤干细胞裸鼠移植瘤生长及微血管生成的影响.[方法] 无血清堵养法从骨肉瘤细胞株MG-3中分离出类肿瘤干细胞建立裸鼠移植瘤模型.30只成瘤裸鼠随机分 Celecoxib 组和对照组,Celcoxib:25 mg/ (kg·d),用药15 d,第27 d处死裸鼠,观察肿瘤体积、抑瘤率,免疫组化技术检测VEGF表达及CD34标记的MVD值.[结果] 分离的骨肉瘤类肿瘤干细胞有致瘤性,可以建立动物模型.Celecoxib抑瘤率为23.2%,Celecoxib组裸鼠移植瘤的体积、VEGF的表达、MVD值均显著低于对照组(P＜0.05).[结论] 骨肉瘤类肿瘤下细胞可以建立裸鼠骨肉瘤移植瘤模型.Celeeoxib可以抑制肿瘤生长,减少移植瘤组织VEGF的表达,减少微血管生成,具有抗血管生成作用.     

放射性碘标记表皮生长因子核素靶向治疗乳腺癌的实验研究

目的　观察放射性核素碘 (13 1I)偶联表皮生长因子 (EGF)对人乳腺癌裸鼠移植瘤的作用 ,探讨其靶向治疗乳腺癌的有效性和可行性。方法　氯胺 T法碘标EGF和人血清白蛋白 ;建立表达表皮生长因子受体 (EGFR)的人乳腺癌荷瘤裸鼠模型 ,36只裸鼠模型分为 6组 ,分别为阴性对照组、阳性对照组、13 1I EGF静脉组、13 1I 白蛋白组、13 1I组和13 1I EGF瘤内注射组 ,每组 6只。观察肿瘤生长增殖情况及放射性核素碘标EGF对正常组织脏器的放射毒副作用。结果　自第 1次给药后第 7天至第2 6天各时间段 ,13 1I EGF静脉和瘤内注射组肿瘤体积与阴性对照组、13 1I组及13 1I 白蛋白组肿瘤体积差异有统计学意义 (P <0 0 1) ;13 1I EGF静脉和瘤内注射组抑瘤率分别为 82 0 %和 80 7% ,与13 1I组和13 1I 白蛋白组比较 (抑瘤率分别为 7 4 9%、6 91% ) ,差异有统计学意义 (P <0 0 1) ;光学显微镜和透射电镜下见13 1I EGF静脉和瘤内注射组肿瘤细胞发生了一系列不可逆损害变化 ;未发现肝、肾和骨髓非特异性放射损伤。结论　经EGF转载的13 1I对人乳腺癌裸鼠移植瘤有明显抑制肿瘤细胞增殖生长的作用 ,无明显毒副作用。     

槲皮素对内分泌耐药乳腺癌三苯氧胺治疗增敏作用的体内研究

目的:探讨槲皮素(QUE)对内分泌耐药乳腺癌三苯氧胺(TAM)治疗的增敏作用。方法:用大剂量TAM冲击法构建TAM耐药乳腺癌细胞株MCF-7/TAM-R并移植裸鼠后,将荷瘤鼠随机分为4组,分别给予溶媒(对照组)、QUE 50 mg/kg 1次/2 d(QUE组)、TAM 5 mg/kg 1次/d(TAM组)、QUE 50 mg/kg 1次/2 d+TAM 5 mg/kg 1次/d(QUE+TAM组)处理,动态观测各组荷瘤鼠的一般情况与瘤体体积的变化,于给药21 d后,处死各组荷瘤鼠,检测瘤体质量及瘤组织ERα、HER-2、pMAPK、MAPK、pAkt、Akt蛋白的表达。结果:给药过程中,QUE+TAM组和QUE组裸鼠摄食减少、体质量减轻,对照组与TAM组裸鼠无明显异常;至第12天开始,QUE+TAM组瘤体生长呈下降趋势,且第21天明显下降(P0.05),其余各组瘤体均呈持续增长。与对照组比较,QUE+TAM组瘤体质量明显减轻(P0.05),而其余两组均无统计学差异(均P0.05);QUE+TAM组和QUE组瘤组织中ERα蛋白高表达,HER-2、pMAPK、pAkt蛋白低表达,而TAM组上述蛋白表达均无明显差异,各组非磷酸化的MAPK、Akt蛋白表达均无明显差异。结论:QUE能恢复内分泌耐药乳腺癌对TAM的敏感性,可能与其下调HER-2及其下游信号pMAPK、pAkt的表达,并上调ERα的表达有关;QUE有潜在的毒副作用,其安全范围及有效剂量有待进一步探讨。     

RNA干扰靶向敲减ObR基因对MCF-7细胞裸鼠移植瘤生长的影响

目的 探讨瘤内注射慢病毒介导的瘦素受体的特异性ObR- siRNA对MCF-7人乳腺癌细胞裸鼠移植瘤生长的影响.方法 建立荷MCF-7人乳腺癌细胞移植瘤裸鼠模型,共30只,随机分为3组:ObR-siRNA慢病毒干预组、NC- siRNA阴性慢病毒对照组和空白对照组,同时在肿瘤部位周围皮下注射重组人瘦素.隔日测量并记录移植瘤的大小,Real-time PCR和Western blotting方法检测ObR的mRNA和蛋白水平的表达,测量肿瘤体积,计算肿瘤抑制率.结果 成功建立了具有高瘦素微环境的敲减ObR基因的荷MCF-7人乳腺癌细胞移植瘤裸鼠模型.ObR- siRNA慢病毒干预组裸鼠移植瘤的体积与NC- siRNA阴性慢病毒对照组和空白对照组差异有统计学意义(P＜0.01,P＜0.01).ObR- siRNA慢病毒干预组中的ObR的mRNA和蛋白水平的表达明显降低,相对NC- siRNA阴性慢病毒对照组和空白对照组差异有统计学意义(P＜0.01,P＜0.01),其抑瘤率为88％.结论 利用慢病毒载体成功将ObR-siRNA导入移植瘤内,且能显著抑制MCF-7细胞裸鼠移植瘤的生长,提示瘦素受体有可能成为乳腺癌基因治疗的靶点.     

三氧化二砷对裸鼠人乳腺癌移植瘤的作用及机制

目的 探讨三氧化二砷(As2O3)对乳腺癌移植瘤的作用及其机制.方法 建立BALB/C-nu/nu裸鼠ER阴性MDA-MB-435s人乳腺癌移植瘤模型,分别以不同浓度的AS2O3、顺铂和0.9%氯化钠注射液(NS)腹腔内注射,观察移植瘤的瘤重及抑瘤率.并用免疫组化检测乳腺癌石蜡标本中PTEN和Casepase7的表达.结果 AS2O3高剂量组、AS2O3低剂量组、顺铂组均能明显抑制裸鼠皮下肿瘤的生长,抑瘤率分别为48.68%、32.80%、66.67%.用药后移植瘤组织巾PTEN和Casepase-7蛋白阳性细胞数显著增多(P＜0.05).结论 AS2O3能明显抑制乳腺癌移植瘤的生长,其作用机制可能是通过上凋PTEN和Caspase-7的表达(P＜0.05),促进细胞的凋亡来抑制人乳腺癌细胞的生长.     

IGF-1R硫代反义寡核苷酸抗人肝癌裸鼠原位移植瘤的研究

目的探讨IGF-1R硫代反义寡核苷酸（IGF-1R APSODN）对人肝癌裸鼠原位移植瘤的治疗效果及毒性作用。方法构建40只人肝癌裸鼠原位移植瘤模型，设溶剂对照组、IGF-1R APSODN（75，50，25mg／kg）三剂量组及氟尿嘧啶阳性对照组，各组裸鼠静脉给药25次，用药期间监测裸鼠体重变化，实验结束后行全血分析，肝肿瘤体积、重量测量，计算抑瘤率，病理组织学检查。重复实验一次。结果两次实验IGF-1R APSODN各剂量组肝肿瘤体积和重量均小于溶剂对照组，其中75mg／kg组抑瘤率分别为76．36％、71．81％，50mg／kg组抑瘤率分别为55．65％、61．74％，25mg／kg组抑瘤率分别为47．72％、50．34％。结论IGF-1R APSODN是一种高效、低毒的抗肝癌生物学药物。     

N-硝基精氨酸甲酯对大肠癌裸鼠移植瘤生长的抑制作用

目的观察一氧化氮合酶(NOS)抑制剂N-硝基精氨酸甲酯(L-NAME)抑制大肠癌生长的体内效应,及其对大肠癌移植瘤相关血管形成的影响。方法将20只BALB/c(nu/nu)裸鼠按随机数字表法均分为对照组和实验组,大肠癌细胞系SL174T种植入裸鼠皮下,形成移植瘤。对照组0.2ml生理盐水灌胃,实验组经口灌入L-NAME,4mg/次,3次/周,共4周。然后记录肿瘤大小,采用免疫组化法检测微血管密度。结果L-NAME对裸鼠大肠癌移植瘤的肿瘤重量抑瘤率为41.36%;肿瘤体积抑瘤率为43.48%。实验组大肠癌组织微血管密度(14.83±2.10)个,明显低于对照组的(21.04±3.11)个,两组间差异有统计学意义(P<0.05)。结论L-NAME具有抑制裸鼠大肠癌肿瘤生长和对抗肿瘤新生血管生成的作用。     

Vitamin D3对乳腺癌荷瘤裸鼠的治疗研究

目的 探讨维生素D3(vitamin D3，VitD3)对乳腺癌裸鼠移植模型的治疗作用。方法 裸鼠皮下接种MCF-7乳腺癌细胞，建立乳腺癌裸鼠移植模型。用VitD3和三苯氧胺(tamoxifen，TAM)给荷瘤裸鼠用药。用药4周后，检测肿块大小、血清钙、磷水平，并用流式细胞仪检测细胞凋亡和细胞周期。结果 VitD3治疗组血清钙水平高于对照组，血磷水平低于对照组，VitD3使肿瘤细胞凋亡增加，并使细胞停留在G0／G1期，与TAM有协同作用。结论 VitD3和TAM可明显诱导乳腺癌细胞凋亡，并阻滞细胞停留在G0／G1期。     

塞来昔布联合奥沙利铂对人结肠癌裸鼠移植瘤生长的抑制作用

目的 观察塞来昔布或联合奥沙利铂对人结肠癌裸鼠移植瘤生长的影响并探讨其机制.方法 用人结肠癌HT-29细胞建立移植瘤模型,将裸鼠随机分为对照组、奥沙利铂组、塞来昔布组、联合用药组.给予相应药物35 d后,取移植瘤组织检测COX-2,VEGF mRNA和微血管密度.结果 塞来昔布组、奥沙利铂组和联合用药组抑瘤率分别为34.94%、30.53%和62.87%.奥沙利铂组COX-2,VEGF表达显著高于对照组(分别P＜0.05).奥沙利铂组微血管密度与对照组比较差异无统计学意义(P＞0.05).塞来昔布组和联合用药组COX-2,VEGF和MVD与对照组比较均显著下降(分别P＜0.05).结论 塞来昔布可抑制人结肠癌裸鼠移植瘤的生长和肿瘤血管生成.塞来昔布增加了奥沙利铂的抗肿瘤效果.     

熊果酸对MCF-7乳腺癌细胞生长的影响

目的 观察熊果酸对MCF-7乳腺癌细胞增殖、凋亡、细胞侵袭及裸鼠移植瘤生长的影响.方法 将1、10、100 μmol/L的熊果酸分别作用于乳腺癌MCF-7细胞12、24、48 h,噻唑蓝(MTT)比色法检测细胞活力变化及生长抑制率,流式细胞术及Transwell小室法检测熊果酸作用48h时MCF-7细胞周期、细胞凋亡率及细胞侵袭力.将乳腺癌MCF-7细胞接种于裸鼠,成瘤后分为生理盐水对照组、熊果酸低剂量组(每日1 mg/kg)、中剂量组(每日5 mg/kg)和高剂量组(每日25 mg/kg),检测5、10、15 d裸鼠移植瘤体积、肿瘤生长抑制率及15 d时肝肾功能、血常规变化.结果 随熊果酸浓度增加及作用时间延长,MCF-7细胞生长及凋亡发生受到显著影响,100 μmol/L熊果酸作用48 h时,细胞生长抑制率为46.0%,生长周期阻滞于G0/G1期,细胞凋亡率为(16.48±2.46)%,细胞侵袭力显著下降.熊果酸组裸鼠移植瘤体积显著小于对照组,15 d时熊果酸高剂量组肿瘤体积为(323.5±33.5)mm3生长抑制率为50.9%.各组肝肾功能和血常规无明显变化.结论 熊果酸可引起体外乳腺癌McF-7细胞增殖抑制、细胞凋亡率增加及细胞侵袭力下降,可显著抑制裸鼠乳腺癌移植瘤生长.     

Human glioblastoma xenografts overexpressing a tumor-specific mutant epidermal growth factor receptor sensitized to cisplatin by the AG1478 tyrosine kinase inhibitor

OBJECT: Activation of signaling by the epidermal growth factor receptor (EGFR) through gene amplification or rearrangement is common in human malignancy, especially in a large fraction of de novo glioblastomas multiforme (GBMs). The most common mutant EGFR, (AEGFR, also known as de2-7 EGFR and EGFRvIII) lacks a portion of the extracellular domain, enhances tumorigenicity in vivo, and causes resistance to the chemotherapeutic drug cisplatin (CDDP). This resistance is due to the suppression of CDDP-induced apoptosis by the constitutively active tyrosine kinase activity of the receptor. The authors have investigated whether inhibition of AEGFR signaling by the tyrosine kinase inhibitor, tyrphostin AG1478, could sensitize tumor xenografts to CDDP and, thereby, enhance its therapeutic efficacy in animals. METHODS: Nude mice were inoculated either subcutaneously or intracerebrally with human GBM cells expressing AEGFR and were then systemically treated with CDDP and/or AG1478. Tumor volumes were monitored and tumor sections were analyzed by using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assays or MIB-1 staining. Expression of AEGFR, but not wild-type EGFR, conferred CDDP resistance to the cells in vivo. Inhibition of receptor signaling by the EGFR-specific tyrosine kinase inhibitor, AG1478. sensitized the xenografts to the cytotoxic effects of CDDP. This combined CDDP/AG1478 treatment significantly suppressed growth of subcutaneous xenografts in nude mice in a synergistic manner (p < 0.01 compared with vehicle control) without causing generalized toxicity, whereas treatments with CDDP or AG1478 alone were ineffective. The synergistic growth suppression by the CDDP/AG1478 combination was not observed in xenografts overexpressing wild-type EGFR or kinase-deficient AEGFR. The combined CDDP/ AG1478 treatment induced tumor growth suppression, which correlated with increased apoptosis and reduced proliferation. This treatment also extended the life span of mice bearing intracerebral xenografts (p < 0.01 compared with controls). CONCLUSIONS: The results of this study may provide the basis for the development of a novel and safe therapeutic strategy for the very aggressive AEGFR-expressing GBM.     

Effects of tamoxifen and somatostatin analogue on growth of human medullary, follicular, and papillary thyroid carcinoma cell lines: tissue culture and nude mouse xenograft studies

The knowledge that (1) the normal thyroid contains somatostatin, (2) polypeptide growth factors influence thyroid cell function, and (3) thyroid cells contain steroid hormone receptors prompted us to add somatostatin analogue No. 201-995 (SMS) (5 ng/ml) and/or tamoxifen citrate (TAM) (5 mumol/L) to 7-day monolayer cultures (50,000 cells/well) of three separate human thyroid carcinoma cell lines: DR081 (medullary), WR082 (follicular), and NPA'87 (papillary). Results, tabulated as cell numbers/well (X10(5) on day 7, revealed that TAM inhibited growth of medullary and follicular cells and that TAM plus SMS inhibited growth of papillary cells. In vivo studies of subcutaneous tumor cell xenografts in nude mice have documented that TAM (5 mg subcutaneous pellet) significantly inhibits the growth of medullary implants. Flow cytometric DNA studies of medullary cell cultures demonstrated a reduced G2 + M phase with TAM treatment. For papillary cell implants, TAM plus SMS (5 micrograms subcutaneously, twice daily) did not suppress tumor growth. All three cell lines were negative for estrogen receptor; addition of estradiol (5 ng/ml) to medullary cell cultures neither stimulated replication nor reversed the inhibitory effects of TAM in vitro. We conclude that (1) TAM slowed the growth of a cell line of human medullary carcinoma, both in vitro and in vivo; (2) this effect was not reversed by estradiol; (3) TAM plus SMS inhibited replication of a papillary carcinoma cell line in vitro, but not in vivo; and (4) TAM alone and TAM plus SMS inhibited replication of cultures of a human follicular thyroid carcinoma cell line. TAM and SMS may be useful in treatment of some human thyroid carcinomas.     

Inhibition of growth of human breast carcinomas in vivo by somatostatin analog SMS 201-995: treatment of nude mouse xenografts

Minced tumor fragments were xenografted into subcutaneous tissue of the lateral thoracic regions of young adult, virgin female nude mice to study the effects of somatostatin analog SMS 201-995 on growth of estrogen-dependent (MCF-7) and estrogen-independent (BT-20) human breast carcinomas. When tumors became palpable (6 to 10 days), mice were assigned randomly to receive either SMS (4 to 50 micrograms) or acetate buffer (0.2 ml) subcutaneously twice a day. For MCF-7, mean tumor volume was significantly lower on day 20 and days 30 through 50 in SMS-treated mice than in controls (p less than 0.05), and tumor doubling time was increased from 13.2 to 19.0 days. Calculated growth increment was significantly lower with SMS than with buffer treatment (1.1 +/- 0.1 vs 1.9 +/- 0.2) (p less than 0.001). For BT-20, mean tumor volume of SMS-treated mice was slightly, but not significantly, lower than that of controls; however, calculated growth increment was significantly lower for SMS treatment (3.2 +/- 0.3 vs 3.9 +/- 0.4) (p +/- 0.001), and tumor doubling time was increased from 4.0 to 5.8 days. For MCF-7, flow cytometric DNA analysis of tumor biopsy samples demonstrated a reduced G2 + M phase with SMS treatment. We conclude that SMS slows the growth of both MCF-7 and BT-20 human breast cancer xenografts in nude mice and that SMS may be clinically useful in the management of patients with breast carcinoma.     

PTEN真核表达质粒对人膀胱癌裸鼠移植瘤的抑制效应

目的:探讨PTEN真核表达质粒对人膀胱癌裸鼠移植瘤的抑制作用。方法:将人膀胱癌细胞株BIU-87裸鼠背部皮下接种,成瘤后于瘤内多点注射重组真核表达质粒pBp-PTEN,设pBp空质粒和生理盐水为对照,观察肿瘤生长情况、PTEN表达的变化。结果:pBp-PTEN治疗组肿瘤变小,PTEN表达阳性率提高,与对照组比较,差异有统计学意义(P<0.05)。结论:PTEN真核表达质粒pBp-PTEN瘤内注射对人膀胱癌裸鼠移植瘤有一定的抑制作用。     

靶向表皮生长因子受体和血管内皮生长因子受体-2治疗多形性胶质母细胞瘤

目的 观察靶向表皮生长因子受体(EGFR)的单克隆抗体C225和靶向血管内皮生长因子受体-2(VEGFR-2)的单克隆抗体DC101单独或联合应用后对人脑多形性胶质母细胞瘤(GBM)裸小鼠脑内移植瘤的疗效.方法 将40只荷瘤鼠随机分为磷酸盐缓冲液(PBS,0.1 ml/20 g·次)、C225(50 mg/kg·次)、DC101(40 mg/kg·次)及C225+DC1O1(50+40 ms/ks·次)等4组,10只/每组,2d1次腹腔用药或PBS,共4次.治疗后观察移植瘤微血管密度(MVD)、体积、形态、增殖和凋亡变化.结果 与对照组比较,DCl01组移植瘤MVD减少了64.O%.体积下降了59.7%,增殖指数下降了53.2%,凋亡指数增加了66.7%(P＜0.05),但肿瘤侵袭性增强;C225组肿瘤侵袭性降低,但对移植瘤MVD、体积、增殖和凋亡无影响(P＞0.05);C225+DC101组移植瘤MVD减少了68.0%,体积下降了62.9%,增殖指数下降了56.4%,凋亡指数增加了75.0%(P＜0.05),肿瘤侵袭性明显下降.结论 DC101抑制血管生成同时增加了肿瘤的侵袭性,而C225能降低肿瘤侵袭性.联合用药不仅抑制GBM移植瘤生长还降低肿瘤侵袭能力.     

Antiestrogens do not counteract the inhibitory effect of estradiol-17β on the growth of the dunning R3327 prostatic adenocarcinoma

The purpose of this study was to investigate if the local effects of estradiol on the Dunning R3327 prostatic adenocarcinoma were estrogen-receptor mediated. All rats with the transplantable Dunning R3327 prostatic adenocarcinoma were castrated on the first day of treatment and were supplemented with daily s.c. injections of testosterone propionate (0.1 mg) during the treatment period, lasting for 6 weeks. The following treatment groups were studied: castration + testosterone supplementation (C + T, control group), C+T and estradiol-17β (50 μg/daily s.c.), C + T and tamoxifen (1 mg twice a week s.c.), and C + T and estradiol-17β in combination with tamoxifen. Tumor volumes were measured every week. At the end of the treatment period, pieces of the tumors were taken for morphological studies and estrogen-receptor analysis. In the groups of rats given tamoxifen treatment no estrogen-receptor binding was detectable in prostatic tumors, but, despite this, tamoxifen did not prevent either the inhibitory effect of estradiol-17β on the tumor growth rate or the estrogen-induced decrease of volume density of prostatic glandular epithelium. In contrast, the estrogen-induced increase of volume density of the stroma was abolished by tamoxifen, suggesting that this effect may be mediated by the estrogen receptor. A morphometrical method for estimating the growth of different tumor compartments is presented. Treatment with estradiol-17β, both with or without combined treatment with tamoxifen, reduced the growth of both the tumor epithelium and stroma. The direct effect of estradiol-17β on the growth and morphology of the Dunning R3327 prostatic adenocarcinoma seemed not to be mediated by the estrogen receptor.     

绿脓杆菌制剂抑制肝细胞肝癌生长转移的实验研究

目的 观察绿脓杆菌制剂对裸鼠肝癌生长和转移的抑制作用.方法 将具有肺转移潜能的人MHCC97L肝癌组织块种植于BALB/c nu/nu雄性裸鼠肝脏,建立转移性人肝癌裸鼠原位模型.荷瘤裸鼠随机分为对照组、绿脓杆菌制剂腹腔注射组及皮下注射组.各组均于种植后第3天开始用药,于种植后第6周末处死动物,HE染色观察肿瘤组织坏死情况,荧光Tunel法检测肿瘤组织细胞凋亡情况,测量肿瘤体积,计算抑瘤率、肺转移灶数目及肺转移率.结果 对照组、绿脓杆菌制剂皮下注射组和腹腔注射组肿瘤体积分别为(3.12±0.85)cm~3、(1.90±0.87)cm~3(P>0.05)和(0.79±0.36)cm~3(P<0.05),肺转移率分别为66.7%、66.7%(P>0.05)和0(P<0.01),平均肺转移灶数目分别为2.40±1.85,1.2±0.75(P<0.05)和0(P<0.01),凋亡指数分别为(1.4±0.37)%、(4.1±1.7)%(P<0.05)和(10.3±0.40)%(P<0.01).与对照组比较,绿脓杆菌制剂皮下注射组和腹腔注射组的抑瘤率分别为39.1%和74.7%,腹腔注射组肿瘤组织坏死明显.结论 绿脓杆菌制剂腹腔注射可明显促进肝癌组织坏死及凋亡并抑制其生长和转移.     

纳米磁小体靶向药囊治疗人胆管癌细胞株移植瘤实验研究

目的:将纳米磁小体靶向药囊(TM5-FUNC)经尾静脉注射入胆管癌荷瘤裸鼠体内,通过埋植在移植瘤内的磁化支架所产生的磁靶向引导作用,TM5-FUNC选择性地聚集于裸鼠胆管癌移植瘤组织内;探讨此装置对人胆管癌的生长抑制作用。方法:建立人胆管癌裸鼠移植瘤模型,将裸鼠随机分成6组,通过尾静脉按分组方案给药,分别将TM5-FUNC 250mg/kg、200mg/kg和150mg/kg组列为高、中、低剂量组。计算各组裸鼠肿瘤体积、抑瘤率和肿瘤生长曲线。处死裸鼠后,将肿瘤标本置电镜下观察肿瘤组织超微结构改变情况。结果:结合内置支架的TM5-FUNC高、中剂量组的移植肿瘤体积在治疗后35d与对照组比较有显著差异(P<0.05);TM5-FUNC低剂量组的肿瘤体积与对照组比较则无明显差异(P>0.05)。其肿瘤的抑制率分别为:高剂量组39.6%、中剂量组14.6%、低剂量组7.9%。从高、中、低剂量TM5-FUNC干预组中可见,随着药物浓度的增高,肿瘤生长越缓,高剂量组肿瘤生长曲线变化最为缓慢,中剂量组次之,其余组曲线变化较为一致。电镜结果显示,TM5-FUNC对荷瘤裸鼠的肿瘤细胞有诱导凋亡作用,且随着药物浓度的增大,细胞凋亡的形态改变越为明显。结论:本课题组所研制的TM5-FUNC新剂型,在内磁场的导向作用下,能有效抑制人胆管癌裸鼠移植瘤的生长。     

Receptor tyrosine kinase inhibition suppresses growth of pediatric renal tumor cells in vitro

PURPOSE: Children who undergo standard therapy for renal tumors are at an increased risk for treatment sequelae such as congestive heart failure, abnormal trunk development, and secondary malignancies. Therefore, research on the use of novel chemotherapeutic agents with fewer side effects is justified. Recent experimental evidence suggests that growth factor receptors such as epidermal growth factor receptor (EGFR) and platelet-derived growth factor receptor (PDGFR) play an important role in growth and development of pediatric renal tumors especially that of Wilms' tumor. In this study we investigated the effects of genistein, AG1478, and AG1295, from the class of growth factor receptor tyrosine kinase (GFR-TK) inhibitors, on proliferation and colonigenic growth of 2 pediatric renal tumor cell lines. METHODS: The authors studied the effect of genistein (broad-spectrum GFR-TK inhibitor), AG1478 (EGFR-specific GFR-TK inhibitor), and AG1295 (PDGFR-specific GFR-TK inhibitor) on proliferation and colonigenic growth of rhabdoid tumor of the kidney and Wilms' tumor cell lines: G-401 and SK-NEP-1, respectively. The effect of genistein at concentrations of 0 to 200 micromol/L, and AG1478 and AG1295 at 0 to 10,000 nmol/L were tested on proliferation by using a growth inhibition assay. Viable cell counts at each concentration were obtained by hemocytometer and trypan blue exclusion, and percent growth inhibition was calculated based on control cultures at the same time-point. As a measure of colonigenic survival, the percent inhibition of colony formation in drug-treated dishes was calculated based on the number of colonies (>50 cells) in control dishes. RESULTS: Genistein at concentrations of 25 and 50 micromol/L inhibited the colonigenic growth of G-401 by 37% and 79% (P = .01 and 5E-06, 2-tailed t test, respectively) and that of SK-NEP-1 by 44% and 74% (P = .0001 and 9.9E-07). The mean percent growth inhibition at the above doses was 57% +/- 7.9% and 96% +/- 0.2% for G-401, and 47% +/- 11.2% and 60% +/- 2.7% for SK-NEP-1. AG1478 at concentrations of 1,000 and 5,000 nmol/L inhibited the colonigenic growth of G401 by 75% and 78% (P = .0005 and 7.38E-06, respectively) and that of SK-NEP-1 by 19% and 40% (P = .02 and .0001). The percent growth inhibition at the mentioned concentrations for G-401 were 53% +/- 9.3% and 63% +/- 6.3%, and for SK-NEP-1 were 55% +/- 14.5% and 65% +/- 20.1%, respectively. AG1295 did not appear to be as effective as AG1478. CONCLUSIONS: This is the first experimental study on the use of GFR-TK inhibitors as a potential treatment for pediatric renal tumors. GFR-TK inhibitors such as genistein occur naturally in soybean foods and have been shown to reach therapeutic levels in blood after consuming a soybean-based diet. Considering the significance of growth factor receptor activity in Wilms' tumor development, inhibition of GFR-TKs should be investigated as effective and potentially nontoxic adjunctive treatment for this childhood tumor. Furthermore, GFR-TK inhibitors may offer an effective alternative to the treatment of commonly fatal rhabdoid tumor of the kidney in children.