20(S)-人参皂苷Rg3对B16黑色素瘤生长的抑制作用

目的 :探讨 2 0 (S) 人参皂苷Rg3对B16黑色素瘤生长的影响。 方法 :体内建立B16实体瘤模型及PCNA免疫组织化学染色 ,体外应用MTT法、生长曲线观察Rg3对B16黑色素瘤生长影响。 结果 :实体瘤模型中 ,Rg3各组瘤重明显低于对照组 (P <0 0 1) ,且Rg3组PCNALI与对照组比较有差异 (P <0 0 5 ) ,随着Rg3给予浓度的增加 ,PCNALI逐渐降低。体外实验观察到Rg3对B16黑色素瘤细胞生长同样有抑制作用。 结论 :Rg3可以明显抑制B16黑色素瘤的生长及增殖活性     

人参皂苷Rg3对食管癌细胞放射增敏作用的初步研究

目的 探讨人参皂苷Rg3（简称Rg3）对食管癌EC109细胞的放射增敏作用。方法采用四甲基偶氮唑盐比色法观察不同浓度的Rg3（10、20、50、100、200、400、600mmol/L）作用EC109细胞24、48及72h的细胞存活率,克隆形成实验及单击多靶模型计算10mmol/L Rg3预处理24h经0、1、3、6和9Gy X射线照射后的辐射增敏比（SER）。根据实验设计分为对照组、单纯照射组及Rg3+照射组,流式细胞仪及Foci焦点形成实验分别检测3组经8Gy X射线照射48h的细胞凋亡率和DNA分子损伤情况。结果在10～600mmol/L范围内,Rg3可降低EC109细胞的存活率,呈剂量和时间依赖性;10mmol/L Rg3处理EC109细胞的SER为1.28。Rg3+照射组的凋亡率为（62.33±4.60）%,高于对照组的（6.46±1.23）%和单纯照射组的（30.68±3.55）%,差异有统计学意义（P＜0.05）;Rg3+照射组的Foci焦点细胞数为（64±12）个,亦高于对照组的（6±3）个和单纯照射组的（32±6）个,差异有统计学意义（P＜0.05）。结论Rg3对食管癌EC109细胞有放射增敏作用,能够抑制细胞活性并诱导细胞凋亡与DNA分子损伤。     

人参皂苷Rg3对小鼠B16黑素瘤细胞侵袭、转移及MMP-9表达的影响

目的:观察人参皂苷Rg3(简称Rg3)对小鼠黑素瘤细胞B16的肺转移、侵袭能力及基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)表达的影响,探讨其抗肿瘤转移的作用机制。方法:体内建立小鼠自发肺转移和实体瘤模型,观察腹腔注射不同剂量的Rg3(对照组给予0.9%氯化钠溶液)后,肺部肿瘤转移灶的数目,并检测实体瘤中MMP-9蛋白的表达情况。采用Boyden小室侵袭实验及免疫细胞化学染色法检测Rg3对肿瘤细胞体处侵袭能力及MMP-9表达的影响。结果:采用不同剂量的Rg3(0.3、1.0和3.0mg/kg)治疗后,小鼠肺部转移灶的数目均较少,肿瘤组织中MMP-9的表达水平降低,与对照组比较差异有统计学意义(P<0.05)。在体外,2.5和5.0μg/mLRg3治疗组中侵袭穿过人工基膜的B16细胞数目明显少于对照组(P<0.01),且5.0μg/mLRg3可抑制肿瘤细胞中MMP-9的表达。结论:Rg3能够抑制小鼠黑素瘤细胞的肺转移,其抗肿瘤转移作用可能是通过降低肿瘤细胞中MMP-9的表达水平及细胞侵袭能力来实现的。     

人参皂苷Rg3的抗肿瘤作用及研究进展

肿瘤是因细胞生长调控机制失控而引起的疾病,严重威胁了人类生命健康。虽然目前有多种疗法用于肿瘤治疗,但是效果均不理想。化疗是目前最常用的治疗方法之一,然而其严重不良反应和产生耐药性影响了化疗药物的治疗效果。人参皂苷Rg3是从人参中分离出来的主要成分之一,其在预防和治疗癌症中起重要作用。人参皂苷Rg3的抗肿瘤机制主要包括诱导肿瘤细胞凋亡、抑制增殖、转移和血管生成,以及解除免疫抑制、促进免疫应答。此外,人参皂苷Rg3与化疗联合可协同改善治疗效果和减少不良反应。本文将近年关于人参皂苷Rg3的文献作一综述,以便更全面地了解人参皂苷Rg3在实体肿瘤中的应用。     

八棱丝瓜蛋白-1对B16细胞的诱导分化作用

目的:测定八棱丝瓜蛋白-1对小鼠黑色素瘤B16细胞的诱导分化作用。方法:以MTT法测定八棱丝瓜蛋白-1对B16细胞的增殖抑制作用,以形态、黑色素含量及细胞周期变化为指标,观察八棱丝瓜蛋白-1诱导B16细胞分化的作用。结果:八棱丝瓜蛋白-1对B16细胞增殖具有明显的抑制作用,呈剂量依赖性和时间依赖性,LF-1对B16细胞作用48和72h的IC50分别为33·29和10·28g/L,并阻止B16细胞由G1期向S期过渡从而停滞于G1期;细胞形态向正常上皮样细胞分化,生长缓慢,平铺不重叠,甚至形成网状结构,胞质内细胞器丰富,可见较多不同成熟期(Ⅰ~Ⅳ期)的黑色素小体;黑色素含量显著增加,2·5、5、10和20mg/L浓度的LF-1处理组的黑色素含量分别为对照组的2·1、4·4、5·9和7·8倍。结论:八棱丝瓜蛋白-1对B16细胞具有明显的诱导分化作用。肿瘤防治杂志,2005,12(20):1521-1524     

八棱丝瓜蛋白-1对B16细胞的诱导分化作用

目的：测定八棱丝瓜蛋白1对小鼠黑色素瘤B16细胞的诱导分化作用。方法：以MTT法测定八棱丝瓜蛋白1对B16细胞的增殖抑制作用，以形态、黑色素含量及细胞周期变化为指标，观察八棱丝瓜蛋白-1诱导B16细胞分化的作用。结果：八棱丝瓜蛋白-1对B16细胞增殖具有明显的抑制作用，呈剂量依赖性和时间依赖性，LF-1对B16细胞作用48和72h的IC50分别为33．29和10．28g／L，并阻止B16细胞由G1期向S期过渡从而停滞于G1期；细胞形态向正常上皮样细胞分化，生长缓慢，平铺不重叠，甚至形成网状结构，胞质内细胞器丰富，可见较多不同成熟期（Ⅰ～Ⅳ期）的黑色素小体；黑色素含量显著增加，2．5、5、10和20mg／L浓度的LF-1处理组的黑色素含量分别为对照组的2．1、4．4、5，9和7，8倍。结论：八棱丝瓜蛋白-1对B16细胞具有明显的诱导分化作用。     

人参皂苷Rg3抑制B16黑色素瘤新生血管生成及其机制的探讨

目的:观察20(S)-人参皂苷Rg3(SPG-Rg3)对B16黑色素瘤血管生成的影响,并探讨其可能的作用机制.方法:体内采用肿瘤诱导血管生成实验,观察SPG-Rg3对B16黑色素瘤血管生成的影响;免疫细胞化学法观察SPG-Rg3对B16黑色素瘤细胞血管内皮细胞生长因子(VEGF)表达的影响;取不同浓度SPG-Rg3作用后的B16黑色素瘤细胞条件培养基(BMCM)作用于人脐静脉内皮细胞,用MTT法、PCNA免疫荧光染色以及迁移实验观察SPG-Rg3对内皮细胞增殖和迁移的作用.结果:在肿瘤诱导血管生成实验中,SPG-Rg3组肿瘤周围的血管数明显少于对照组,P＜0.01;SPG-Rg3作用组BMCM的促内皮细胞增殖和迁移作用均明显弱于无SPG-Rg3作用组,且SPG-Rg3(5 μg/mL)降低了B16黑色素瘤细胞VEGF的表达,P＜0.01.结论:SPG-Rg3可明显抑制B16黑色素瘤的血管生成,且可能通过降低肿瘤细胞分泌VEGF来抑制肿瘤细胞对血管内皮细胞增殖和迁移的促进作用,从而达到抑制肿瘤新生血管形成的作用.     

人参皂甙Rg3诱导细胞凋亡作用的研究

目的观察人参皂甙Rg3对小鼠肝癌淋巴结转移模型中肿瘤细胞凋亡的诱导作用，探讨人参皂甙Rg3抗肿瘤淋巴结转移的机制。方法建立小鼠肝癌淋巴结转移模型；利用免疫组化方法观察原发瘤及转移瘤各组（Rg3预防组，Rg3治疗组，Rg3与顺铂联合治疗组，顺铂治疗组及对照组）中肿瘤细胞的凋亡。结果在原发和转移瘤中，对照组Bcl-2高表达，Bax低表达，Bcl-2在人参皂甙Rg3预防组、Rg3治疗组、Rg3与顺铂联合治疗组，顺铂治疗组中是低表达，明显低于对照组（P〈0．05），Bax明显高于对照组（P〈0．05）。结论人参皂甙Rg3抗肿瘤细胞淋巴结转移的作用与诱导细胞凋亡有关。     

三苯氧胺联合人参皂苷Rg3抑制乳腺癌血管生成的研究

目的：研究三苯氧胺（tamoxifen，TAM）联合人参皂苷R0对乳腺癌肿瘤血管生成的影响，并观察其抑瘤效果。方法：以荷MCF-7乳腺癌裸鼠为模型，分别给予TAM、雌二醇（estradiol，E2）、人参皂苷Rg3、TAM联合人参皂苷Rg3及9％氯化钠溶液灌胃，观察肿瘤生长情况，免疫组织化学染色计数肿瘤微血管密度（microvascular density，MVD），PT-PCR法检测血管内皮生长因子（vascular endothelial growth factor，VEGF）表达。结果：E2处理组的肿瘤生长迅速，而TAM治疗组肿瘤生长受到抑制，合用人参组皂苷Rg3时抑瘤效果更显著（P〈0．05）。与9％氯化钠溶液对照组相比，E2组MVD升高，而TAM组和人参皂苷Rg3组的MVD及VEGF mRNA表达均下降，二者合用时MVD和VEGF mRNA表达下降更明显（P〈0．05）。结论：TAM可抑制乳腺癌肿瘤血管生成，与人参皂苷Rg3联合应用显示出抗血管生成协同作用，可有效抑制乳腺癌肿瘤生长。     

人参皂苷Rg3对人宫颈癌Hela细胞的诱导凋亡作用

目的:探讨人参皂苷Rg3对人宫颈癌Hela细胞的诱导凋亡作用。方法:培养人宫颈癌Hela细胞,不同浓度Rg3处理,HE染色观察细胞形态学改变,CCK-8、流式细胞仪检测细胞的凋亡。结果:Rg3作用后Hela细胞出现了典型的凋亡形态学变化,包括细胞皱缩、细胞核内染色体浓缩。10、20、40、80μg/ml人参皂苷Rg3处理48小时后Hela细胞的平均生存率为76.3%、57.1%、50.0%、29.8%;流式细胞术(FCM)显示肿瘤细胞凋亡率为16.4%、37.0%、50.0%、69.8%,表明肿瘤细胞在人参皂苷Rg3诱导下发生凋亡。结论:Rg3能抑制Hela细胞增殖,诱导其凋亡,其诱导凋亡作用呈剂量依赖性。     

Identification of iGb3 and iGb4 in melanoma B16F10-Nex2 cells and the iNKT cell-mediated antitumor effect of dendritic cells primed with iGb3

Background  

CD1d-restricted iNKT cells are protective against murine melanoma B16F10-Nex2 growing subcutaneously in syngeneic C57Bl/6 mice as inferred from the fast tumor development in CD1d-KO in comparison with wild type animals. CD1d glycoproteins are related to the class I MHC molecules, and are involved in the presentation, particularly by dentritic cells (DC), of lipid antigens to iNKT cells. In the present work we attempted to identify the endogenous lipid mediator expressed in melanoma cells inducing such immunesurveillance response and study the possibility of protecting animals challenged with tumor cells with lipid-primed DC.     

Cytotoxic effect of RA233 on hypoxic B16 melanoma cells in vitro

The pyrimido-pyrimidine derivative RA233 was found to selectively kill cultured mouse B16 melanoma cells after prolonged hypoxia. At the optimum cytotoxic concentration (100 microM), RA233 reduced cell clonogenicity by about 80% when administered during long-term hypoxia of 4 days. Comparable cytotoxicity was also evident when RA233 was present only during re-oxygenation following 4 days of hypoxia. RA233 treatment during both hypoxia and re-oxygenation resulted in the greatest cytotoxicity, with only about 1% of cells surviving such treatment. By contrast, the hypoxic cell sensitizer misonidazole was cytotoxic only when administered during hypoxia. RA233 appears to be a unique hypoxic cell sensitizer that kills long-term hypoxic tumor cells principally during re-oxygenation.     

Antiangiogenic effect of capecitabine combined with ginsenoside Rg3 on breast cancer in mice

Capecitabine is a novel fluoropyrimidine carbamate, which has a broader spectrum of antitumor activity than other fluoropyrimidines, such as 5-FU, DFUR, or UFT; it has proved effective over a wide dose range. Recent research has suggested that frequent administration of lower doses of certain chemotherapeutic drugs might enhance their antiangiogenic effect. The present study investigated the antiangiogenic effect of capecitabine on breast cancer. In order to augment its efficacy, we combined capecitabine chemotherapy with ginsenoside Rg3. Our results indicate that a metronomic regimen of capecitabine inhibited angiogenesis in breast cancer, and its antiangiogenic effects may be further enhanced by the concurrent administration of ginsenoside Rg3. As an antiangiogenic method, this regimen presented better antitumor effects, less toxicity, and reduced susceptibility to drug resistance.     

In vitro and in vivo enhancement of vincristine antitumor activity on B16 melanoma cells by calcium antagonist flunarizine

Flunarizine, a calcium antagonist commonly employed in therapy for vascular diseases, enhances the in vitro and in vivo antitumor activity of vincristine on B16 melanoma cells. In the presence of flunarizine higher intracellular levels of vincristine were observed in vitro and for a longer time. B16 melanoma bearing mice treated with both the drugs presented a median survival time that was significantly longer than that of the controls. The possible mechanism of the enhancement is herein discussed.     

Combined effect of cisplatin and caffeine on murine B16-BL6 melanoma cells

Combined effect of cisplatin and caffeine on murine B16-BL6 melanoma cells was studied. Synergistic inhibition of the cell growth was observed when caffeine (2 mM) was added continuously after one hour exposure of cisplatin. On the other hand, when caffeine was added before one hour exposure of cisplatin or one hour simultaneous exposure with cisplatin, synergistic effect was not shown. In the analysis of DNA histogram obtained from flow cytometry, S and G2/M accumulation was observed by the treatment of cisplatin and that accumulation was reduced by the combination of cisplatin and caffeine. From this findings, it was suggested that caffeine would inhibit DNA repair process. Furthermore, according to morphological studies with hematoxylin-eosin stain and Fontana-Masson stain, the addition of caffeine alone resulted in mild swelling of melanoma cells and the decrease of nuclear-cytoplasmic ratio. The combination of cisplatin and caffeine caused marked swelling of melanoma cells and remarkable increase of dendrite-like processes. Melanogenesis was also enhanced by the addition of these two drugs. Many matured melanosomes, increases of mitochondria, Golgi's apparatus and endoplasmic reticula were observed by the use of electron microscope. These findings implied that the combination of cisplatin and caffeine induced a differentiation of murine melanoma cells.     

沉默pim-3基因对黑色素瘤细胞B16的抑制作用

目的:观察靶向沉默原癌基因pim-3对小鼠黑色素瘤细胞B16的抑制作用.方法:用脂质体将特异性靶向沉默pim-3载体(pim-3-shRNA)和pin-3过表达载体(pim-3-MIGR1)转染到黑色素瘤细胞B16中,荧光定量PCR和Western blotting法检测pim-3 mRNA和蛋白表达的变化,Annexin V/PI染色和TUNEL染色检测细胞凋亡情况,荧光定量PCR和Western blotting法检测p-Bad、Bcl-2、Bcl-xl、Bax、caspase-3等凋亡相关分子表达的变化,MTT法检测细胞增殖情况,流式细胞术检测细胞周期,RTCAxCELLigence系统检测细胞迁移情况,Transwell侵袭实验检测细胞侵袭的变化.结果:转染pim-3-shRNA组B16细胞的pim-3mRNA及蛋白表达较对照组明显下降(P＜0.05),转染pim-3过表达质粒组B16细胞的pim-3 mRNA及蛋白表达较对照组显著增加(P＜0.05).转染pim-3-shRNA后,B16细胞的凋亡明显增加,在此基础上共转pim-3过表达质粒后则明显逆转沉默pim-3所引起的凋亡(p＜0.05);进一步检测发现沉默pim-3明显下调凋亡相关分子p-Bad、Bcl-2、Bcl-xl的表达,上调Bax的表达,最终引起caspase-3活性增加(P＜0.05).沉默pim-3后B16细胞增殖和迁移受到抑制(P.＜0.05),而细胞周期无明显变化.结论:靶向沉默pim3基因能促进黑色素瘤细胞的凋亡,抑制细胞的增殖和迁移,提示pim-3基因可以作为黑色素瘤基因治疗的新靶点.     

Antitumor effect of B16 melanoma cells genetically modified with the angiogenesis inhibitor rnasin

The growth of new blood vessels is an essential condition for the development of tumors with a diameter greater than 1-2 mm and also for their metastatic dissemination. RNasin, the placental ribonuclease inhibitor, is known to have antiangiogenic activity through the inhibition of angiogenin and basic fibroblast growth factor. Nevertheless, the administration of the recombinant form of a protein poses several limitations; as a result, we have studied the antitumor effect of RNasin in a murine gene therapy model. RNasin cDNA was subcloned into the pcDNA3 expression vector, and the resulting recombinant plasmid was used to transfect the B16 murine melanoma cell line. An RNasin inverted construction was used as control. Mice intravenously injected with clones expressing RNasin showed a significant inhibition of tumor metastatic progression with respect to control groups (P<.001) and survived longer (P<.001). Tissue sections from RNasin-expressing cell tumors showed a lower number of blood vessels when compared to tissue sections from mice lungs that had been inoculated with control cell lines. The results of these experiments show that the genetic modification of tumor cells with RNasin cDNA yields a significant antitumor effect, and suggest that this effect is at least partially the result of angiogenesis inhibition.     

肝脂素对小鼠S37肉瘤与B16黑色素瘤的抗肿瘤作用

目的：探讨肝脂素（Heplipin)对S37和B16两种肿瘤细胞移植瘤的生长抑制作用。方法：小鼠S37肉瘤和B16黑色素瘤移植瘤在高、中、低剂量（每天分别为3000、1800、1100mg/kg)肝脂素胃饲为治疗组,环磷酰胺（CTX）20mg(kg／天）和生理盐水作为两对照组。比较抑瘤率、癌周边部血管数、总肿瘤面积中坏死区所占比例,结果：肝脂素对S37肉瘤和B16黑色素瘤的抑瘤分别为大剂量：75．1％和47．0％、中剂量：69．7％和32．1％、小剂量：61．7％和13．0％,显示出明显的剂量效关系,与生理盐水对照组相比有显著性差异。在S37肉瘤荷瘤小鼠的实验中,组织学观察发现,肝脂素用药组肿瘤周边部单痊面积血血管数明显少于阴性对照组和CTX组;肝脂素治疗组的肿瘤坏死面积明显大于对照组,结论：肝脂素可明显抑制小鼠S37肉瘤和B16黑色素瘤的生长,其抗肿瘤机制可能与抑制肿瘤生长血管的生成,导肿瘤缺血坏死有关。     

Suramin augments the antitumor and antimetastatic activity of pentoxifylline in B16F10 melanoma

Rapid tumor growth and metastasis are 2 major problems associated with treatment of malignant melanoma. Therefore, drugs that can intervene these processes are of clinical importance. Pentoxifylline (PTX), a methyl xanthine derivative, has been shown to inhibit B16F10 melanoma tumor growth and metastasis. We hypothesized that suramin when combined with PTX enhances its antineoplastic effects, which we have examined using the B16F10 mouse melanoma model. Suramin in simultaneous or sequential combination potentiated the cytotoxic effects of PTX on B16F10 cells. PTX arrested cells in the G0-G1 phase and suramin augmented the effects. Both the drugs inhibited F10 adhesion to laminin, matrigel and collagen type IV and showed enhanced inhibition in combination The combination also demonstrated significantly higher inhibition in cell motility (p = 0.002) and invasion through matrigel (p = 0.005) as compared to the single agents. Suramin synergized with PTX in its effects on secretion of MMP-9 gelatinase. DBA2/J mice implanted with intradermal B16F10 tumor were used as a model to study tumor growth. Animals were intratumorally treated with 50 mg/kg of PTX, 10 mg/kg of suramin and their combinations. Simultaneous administration of the drugs inhibited tumor growth by 5- to 6-folds. Tumor growth was completely blocked in sequential regimen with regression in some cases. The number and size of metastatic nodules on lung was also reduced significantly by the combination treatment. In conclusion, the novel combination of PTX and suramin has synergistic antitumor and antimetastatic activity in B16F10 melanoma and may be a promising approach in treatment of patients suffering from malignant melanoma.