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。     

PCNA、P16、MMP9在人参皂甙Rg3抗肝癌淋巴道转移中的表达及意义

目的 :通过检测PCNA、P16及MMP 9在人参皂甙Rg3抗荷瘤小鼠淋巴道转移中的表达 ,探讨人参皂甙Rg3抗肿瘤作用的机制。方法 :建立小鼠肝癌淋巴道转移模型 ,应用免疫组织化学方法检测PCNA、P16及MMP 9在各实验组 (Rg3预防组、Rg3治疗组、Rg3+DDP合用组、DDP阳性对照组和生理盐水阴性对照组 )的原发瘤及相应转移瘤 (淋巴结 )中的表达水平。结果 :应用人参皂甙Rg3组较未用药组 ,PCNA及MMP 9在原发瘤的表达减少 ,P16的表达增加 ,差异显著 (P <0 0 0 1) ;而在转移瘤中的变化不明显。结论 :人参皂甙Rg3抗淋巴道转移作用的机制与上调P16表达及下调PCNA和MMP9表达有关。     

人参皂甙Rg3对荷瘤及环磷酰胺化疗小鼠黏膜免疫力影响

[目的]观察人参皂甙Rg3对肿瘤化疗肠道黏膜损伤的治疗作用及其黏膜免疫机制。[方法]运用给小鼠原位接种C-26结肠癌并重复灌胃环磷酰胺建立肿瘤化疗肠道黏膜损伤的模型，观察人参皂甙Rg3对瘤重、抑瘤率及PP结面积影响，分离肠上皮内淋巴细胞，固有层淋巴细胞及PP结内的淋巴细胞，用免疫荧光染色及流式细胞仪分析黏膜免疫各部位T淋巴细胞亚群的变化。[结果]环磷酰胺组瘤重显著降低，抑瘤率约为37.6%,而环磷酰胺加Rg3组瘤重与环磷酰胺组比较差异无显著性。荷瘤及环磷酰胺都可显著降低PP结而积，而人参皂甙Rg3可明显减轻环磷酰胺对PP结面积的影响（P＜0.01）。人参皂甙Rg3可使荷瘤和环磷酰胺作用下显著减少的固有层淋巴细胞中的CD4^＋细胞百分比显著增加（P＜0.05），但对CD8^＋细胞百分比无明显影响。[结论]荷瘤及环磷酰胺可明显抑制肠道黏膜免疫功能，而人参皂甙Rg3能明显改善这种黏膜免疫抑制状态。人参皂甙Rg3对黏膜免疫的调节作用是扶正中药发挥作用的重要途径之一。     

人参皂甙Rg3在小鼠肝癌淋巴结转移模型中诱导细胞凋亡的作用

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;光镜和透射电镜下观察各组(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)中原发瘤及转移瘤组织的形态学结构改变,并通过流式细胞仪分析肿瘤细胞的凋亡.结果:Rg3预防组及治疗组电镜下(5份样品)可见较多细胞凋亡小体的形成(5/5,4/5);顺铂治疗组的形态改变以细胞破坏为主,凋亡的细胞较少;联合治疗组细胞的凋亡和坏死程度相当.流式细胞学检测分析结果为:Rg3预防组、治疗组及联合治疗组均见细胞凋亡的特征峰(5/5),而顺铂治疗组为1/5,对照组未见凋亡峰(0/5).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.     

人参皂甙Rg3抑制小鼠肝癌淋巴道转移作用及其对免疫功能的影响

目的：观察人参皂甙Rg3抑制小鼠肿瘤生长及抗淋巴结转移的作用及对免疫功能的影响。方法：以Hca-F25//6A3-F（F）接种于615近交系小鼠，建立肝癌淋巴道转移动物模型，分为5组：Rg3预防组（接种肿瘤前给Rg3）、Rg3治疗组（Rg3）、阳性对照组（PDD）、联合治疗组（Rg3＋PDD）和阴性对照组（生理盐水），比较各组的原发瘤抑制率和转移淋巴结抑瘤率，并通过流式细胞仪方法分析各组免疫指标CD4／CD8的变化。结果：联合治疗组原发瘤的抑瘤率明显提高，与阴性对照组相比具有统计学上明显差异（P〈0．05）；淋巴结转移抑制率含Rg3组均较阴性对照组高；Rg3预防组和Rg3治疗组CD4／CD8比值与阴性对照组相比升高，具有统计学上明显差异（P〈0．05）。结论：人参皂甙Rg3具有明显的抗肿瘤作用，可以增强化疗药物PDD的抗癌作用，及抑制淋巴道转移作用，并提高荷瘤小鼠免疫功能。     

丹参注射液对lewis肺癌小鼠移植瘤生长转移及VEGF表达的影响

目的:观察不同浓度丹参注射液对lewis肺癌小鼠移植瘤生长转移及肿瘤血管内皮细胞生长因子表达的影响.方法:将荷lewis肺癌的C57BL小鼠,分别腹腔注射不同浓度的丹参注射液[(5,10,20,40,80)g/kg]、环磷酰胺(CTX)、生理盐水,检测小鼠的肺部转移灶数和移植瘤的体积、重量及血管内皮生长因子(VEGF)的表达.结果:丹参各剂量组对荷瘤小鼠瘤重均有抑制作用,在实验范围内无明显的剂量依赖关系,与生理盐水组比较,丹参各剂量组差异无显著性(P>0.05).CTX组对荷瘤小鼠瘤重抑制较明显,与生理盐水组比较,差异有显著性(P<0.05).丹参各剂量组对荷瘤小鼠肿瘤体积有不同程度的抑制,但无明显的剂量依赖关系,与生理盐水组比较,丹参各组差异无显著性(P>0.05).CTX组对荷瘤小鼠肿瘤体积抑制较明显,与生理盐水组比较,差异有显著性(P<0.01).丹参各剂量组的肺转移灶数目均低于生理盐水组,但差异无显著性(P>0.05),CTX组肺转移灶数目低于生理盐水组,差异有显著性(P<0.01).丹参各剂量组对荷瘤小鼠移植瘤VEGF的表达与生理盐水组比较,差异无显著性(P>0.05),CTX组移植瘤VEGF的表达与生理盐水组比较,差异无显著性(P>0.05).丹参各剂量组对荷瘤小鼠脾、胸腺指数的影响与生理盐水组比较,差异无显著性(P>0.05),CTX组脾、胸腺指数与生理盐水组比较,差异无显著性(P>0.05).结论:不同浓度的丹参注射液对荷瘤小鼠的瘤重、瘤体积及肺部转移灶无明显的抑制或促进作用,对肿瘤转移相关因子VEGF的表达无影响.     

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。     

人参皂苷Rg3对小鼠肺腺癌移植瘤生长抑素受体表达的调控及意义

目的:探讨人参皂苷Rg3对肺腺癌小鼠移植瘤生长和转移的抑制作用及其可能的机制.方法:接种Lewis细胞构建高转移肺腺癌小鼠移植瘤模型,随机分为对照组(0.9％氯化钠溶液)、顺铂(cisplatin,DDP)组和人参皂苷Rg3组;肿瘤细胞接种后4d给予相应药物干预,接种后24 d处死小鼠,剥离皮下肿瘤并取出肺组织,计算抑瘤率和肺表面结节转移抑制率;应用免疫组织化学法检测移植瘤组织中生长抑素受体( somatostatin receptor,SSTR)、血管内皮生长因子(vascular endothelial growth factor,VEGF)和增殖细胞核抗原(proliferation cell nuclear antigen,PCNA)的表达水平,TUNEL法检测移植瘤中肿瘤细胞的凋亡情况.结果:DDP组和人参皂苷Rg3组的抑瘤率分别为39.20％和54.86％ (P＜0.01).DDP组和人参皂苷Rg3组的肺表面结节转移抑制率分别为30.25％和58.57％,差异有统计学意义(P＜0.05).人参皂苷Rg3组中SSTR的表达水平和肿瘤细胞凋亡指数高于对照组和DDP组(P＜0.01),人参皂苷Rg3组中VEGF的表达水平和PCNA增殖指数低于对照组和DDP组(P＜0.01,P＜0.05).结论:人参皂苷Rg3对肺腺癌小鼠移植瘤的生长和转移具有明显抑制作用,其机制可能与SSTR的过表达有关.     

小剂量化疗联合人参皂甙Rg3抑制小鼠Lewis肺癌血管生成作用实验研究

背景与目的目前发现一些细胞毒性化疗药物小剂量、短间隙、持续给药可表现出明显的抗肿瘤血管生成作用，从而抑制肿瘤生长和转移，被称作“抗肿瘤血管生成化疗（anti-angiogenic chemotherapy）”。近来从中药人参中提制的人参皂甙Rg3也被证实具有抑制肿瘤血管生成的作用。本研究的目的是观察小剂量吉西他滨和人参皂甙Rg3联合治疗对小鼠Lewis肺癌血管生成的抑制作用，以及对小鼠生存质量的影响。方法建立小鼠Lewis肺癌模型，分别给予小剂量吉西他滨、人参皂甙Rg3以及二者的联合治疗。利用彩色多谱勒超声、免疫组化技术等观察和比较各治疗组的肿瘤血管生成和肿瘤生长情况。结果人参皂甙Rg3和吉西他滨联合治疗组小鼠有较好的生存质量。彩色超声多谱勒以及免疫组化结果发现联合治疗组比单药治疗组具有更高的肿瘤坏死率和更强的抗肿瘤血管生成作用。结论小剂量吉西他滨与人参皂甙Rg3联合治疗可能具有协同抑制肿瘤血管生成的作用，从而取得协同抗肿瘤效应，同时可维持较好的生存质量。     

人参皂苷Rg3对小鼠Lewis肺癌生长和转移的抑制作用及其与肿瘤相关巨噬细胞的关系

目的研究人参皂苷Rg3(ginsenoside Rg3,Rg3)对肺腺癌小鼠异体移植瘤生长和转移的抑制作用及其相关机制。方法构建高转移肺腺癌小鼠移植瘤模型,将24只接种高转移性Lewis肺癌的C57BL/6J小鼠随机分成3组：对照组、顺铂(Cisplatin,DDP)组和Rg3组。给予相应药物干预,于接种后24日处死各组小鼠,剥离皮下肿瘤,取出双肺,计算肺转移发生情况,免疫组织化学检测皮下移植瘤中肿瘤相关巨噬细胞（tumor-associated macrophages,TAMs）和微血管密度（microvessel density,MVD）的表达并定量分析。结果DDP组、Rg3组抑瘤率分别为39.20%、54.86%,与对照组比较,瘤重差异有统计学意义（P<0.01）,肺表面结节转移抑制率分别为30.25%,58.57%,与对照组比较差异有统计学意义（P<0.01）,对照组、DDP组和Rg3组TAMs分别为(47.21±12.06)、(45.67±11.90)、(16.11±6.32),MVD分别为(24.57±4.59)、(23.52±4.74)、(14.17±2.43)。结论Rg3对肺腺癌移植瘤的生长和转移具有抑制作用,其机制可能与其抑制肿瘤内TAMs在肿瘤基质中浸润及抑制微血管生成有关。     

Research on the antitumor effect of ginsenoside Rg3 in B16 melanoma cells

Ginsenoside Rg3 is an effective chemical component extracted from the red Panix. The experiment demonstrated that it might effectively inhibit proliferation and metastasis of tumor cells. The exact molecular mechanism of Rg3 remains unclear so far. To further explore the antitumor function of Rg3, we investigated the in-vitro and in-vivo activity of Rg3 in the treatment of B16 melanoma cells, derived from C57BL/6 mouse, capable of forming tumor colonies in the lungs following intravenous injection. Cell proliferation was measured by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide assay. Morphological changes of cells were observed by staining with Giesma and Hoechst 33258. Cell cycle and apoptosis rate were analyzed by flow cytometry. The expression of caspase-3 and bcl-2 in cells was detected by immunocytochemistry and western blot analysis. We found that Rg3 could inhibit cell proliferation, regulate cell cycle, and induce cell apoptosis in vitro. B16 melanoma-bearing mice were used to evaluate in vivo the antitumor activity of Rg3. Mice that were injected with Rg3 showed significant inhibition of the tumor metastasis with lighter lung weight, lower density of microvessels, fewer metastasis nodules, and longer survival time than those in the control group (P<0.001). In conclusion, the results reveal that antitumor metastasis of Rg3 is also associated with inducing apoptosis, regulating cell cycle, and blocking angiogenesis in addition to inhibiting proliferation. This research might supply valuable data for chemotherapy with Rg3 in melanoma. Rg3 would turn out to be an anticancer drug with promising prospects.     

小剂量X线全身照射对不同荷瘤时间小鼠的抗肿瘤转移效应

采用肿瘤血道转移模型，选用Ｂ１６黑色素瘤和Ｌｅｗｉｓ肺癌（ＬＬＣ）两种类型细胞，就小剂量Ｘ线全身照射对不同荷瘤时间小鼠的抗肿瘤转移效应进行研究。结果发现，于静脉注射Ｂ１６黑色素瘤或ＬＬＣ细胞后２４ｈ（Ｂ组）或第１３天（Ｃ组）接受７５ｍＧｙＸ线全身照射小鼠的肿瘤肺转移结节数明显低于未照射组（Ａ组）（Ｐ＜０．０５～０．００２）。比较不同照射时间的抗肿瘤转移效果时发现，静注Ｂ１６黑色素瘤的Ｂ组肺转移结节数仅为Ｃ组的５９％，而ＬＬＣ的Ｂ组则为其Ｃ组的７１％。检测注射Ｂ１６黑色素瘤第１４天各组小鼠的免疫功能时发现，Ｂ、Ｃ两组ＮＫ、ＬＡＫ细胞活性及脾细胞对ＩＬ－２反应性明显高于Ａ组（Ｐ＜０．０５～０．００１）。上述结果提示：小剂量Ｘ线全身照射提高荷瘤小鼠的免疫功能可能是其抗肿瘤转移作用的主要机制之一。     

Augmentation of metastasis formation by thioglycollate-elicited macrophages

Inoculation of thioglycollate-elicited peritoneal exudate cells (PEC) into C57BL/6 mice reduced the rate of lung clearance of several intravenously (i.v.) injected murine tumor cells, and increased by up to 100-fold the number of artificially-induced metastatic lung nodules produced by the i.v. injection of B16 melanoma or Lewis lung carcinoma (3LL) tumor cells. Maximum effects were observed when PEC were injected either before, or shortly after, tumor cells. Modulation of lung clearance or metastasis formation was observed only with PEC and not with a variety of other cells, such as splenocytes, thymocytes, P815 mastocytoma cells, or several macrophage-like cell lines (PU5-1.8 and IC-21). Lysates of PEC were as efficient in reducing lung clearance and augmenting metastasis formation as were intact viable PEC. Lysates of other cell types, including P815 and the macrophage-like cell lines, were unable to produce these effects. PEC populations, enriched for macrophages by adherence to plastic or by percoll density gradient sedimentation, also increased the number of B16-induced artificial metastasis, implicating the macrophage as the cell responsible for these observations.     

Natural killer T cell ligand alpha-galactosylceramide inhibited lymph node metastasis of highly metastatic melanoma cells.

The role of natural killer T (NKT) cells in the prevention of multiple tumor metastasis was examined. The i.v. inoculation of a highly metastatic subline of B16-BL6 (B16-BL6-HM) melanoma cells resulted in the formation of metastatic nodules in lymph nodes in addition to lung, intrapleural cavity, and ovary. However, treatment of the mice with the NKT cell ligand alpha-galactosylceramide (alpha-GalCer) three times from 1 day after B16-BL6-HM melanoma inoculation caused a significant inhibition of multiple metastasis. Lymph node metastasis of B16-BL6-HM was almost completely blocked by alpha-GalCer treatment. This antimetastatic effect of alpha-GalCer was abolished in NKT cell-deficient mice. These results suggest that alpha-GalCer-activated NKT cells played a critical role in the prevention of lymph node metastasis of melanoma cells.     

Natural Killer T Cell Ligand α-Galactosylceramide Inhibited Lymph Node Metastasis of Highly Metastatic Melanoma Cells

The role of natural killer T (NKT) cells in the prevention of multiple tumor metastasis was examined. The i.v. inoculation of a highly metastatic subline of B16-BL6 (B16-BL6-HM) melanoma cells resulted in the formation of metastatic nodules in lymph nodes in addition to lung, intrapleural cavity, and ovary. However, treatment of the mice with the NKT cell ligand α-galactosylceramide (α-GalCer) three times from 1 day after B16-BL6-HM melanoma inoculation caused a significant inhibition of multiple metastasis. Lymph node metastasis of B16-BL6-HM was almost completely blocked by α-GalCer treatment. This antimetastatic effect of α-GalCer was abolished in NKT celldeficient mice. These results suggest that α-GalCer-activated NKT cells played a critical role in the prevention of lymph node metastasis of melanoma cells.     

Gene silencing of beta-catenin in melanoma cells retards their growth but promotes the formation of pulmonary metastasis in mice

Altered expression of beta-catenin, a key component of the Wnt signaling pathway, is involved in a variety of cancers because increased levels of beta-catenin protein are frequently associated with enhanced cellular proliferation. Although our previous study demonstrated that gene silencing of beta-catenin in melanoma B16-BL6 cells by plasmid DNA (pDNA) expressing short-hairpin RNA targeting the gene (pshbeta-catenin) markedly suppressed their growth in vivo, gene silencing of beta-catenin could promote tumor metastasis by the rearranging cell adhesion complex. In this study, we investigated how silencing of beta-catenin affects metastatic aspects of melanoma cells. Transfection of B16-BL6 cells with pshbeta-catenin significantly reduced the amount of cadherin protein, a cell adhesion molecule binding to beta-catenin, with little change in its mRNA level. Cadherin-derived fragments were detected in culture media of B16-BL6 cells transfected with pshbeta-catenin, suggesting that cadherin is shed from the cell surface when the expression of beta-catenin is reduced. The mobility of B16-BL6 cells transfected with pshbeta-catenin was greater than that of cells transfected with any of the control pDNAs. B16-BL6 cells stably transfected with pshbeta-catenin (B16/pshbeta-catenin) formed less or an equal number of tumor nodules in the lung than cells stably transfected with other plasmids when injected into mice via the tail vein. However, when subcutaneously inoculated, B16/pshbeta-catenin cells formed more nodules in the lung than the other stably transfected cells. These results raise concerns about the gene silencing of beta-catenin for inhibiting tumor growth, because it promotes tumor metastasis by reducing the amount of cadherin in tumor cells.     

苦参碱衍生物M19对于肿瘤细胞转移的抑制作用

目的:探讨苦参碱衍生物M19对于肝细胞癌和黑色素瘤细胞转移的抑制作用。方法:肝癌细胞Hep3B、MHCC-LM3和黑色素瘤细胞B16F10经M19处理后,检测肿瘤细胞体外迁移能力。建立C57BL/6小鼠黑色素瘤B16F10和裸鼠肝细胞癌MHCC-LM3转移模型,经M19体内给药后,观察肿瘤细胞肝、肺转移情况。结果:M19能浓度依赖性地抑制肝癌细胞和黑色素瘤细胞迁移。M19体内给药使B16F10细胞在小鼠肺的转移减少,且还能抑制MHCC-LM3细胞在裸鼠肺和肝的转移。结论:M19能在体内外抑制肝细胞癌和黑色素瘤细胞的转移。     

A novel function of junctional adhesion molecule-C in mediating melanoma cell metastasis

Hematogenous dissemination of melanoma is a life-threatening complication of this malignant tumor. Here, we identified junctional adhesion molecule-C (JAM-C) as a novel player in melanoma metastasis to the lung. JAM-C expression was identified in human and murine melanoma cell lines, in human malignant melanoma, as well as in metastatic melanoma including melanoma lung metastasis. JAM-C expressed on both murine B16 melanoma cells as well as on endothelial cells promoted the transendothelial migration of the melanoma cells. We generated mice with inactivation of JAM-C. JAM-C(-/-) mice as well as endothelial-specific JAM-C-deficient mice displayed significantly decreased B16 melanoma cell metastasis to the lung, whereas treatment of mice with soluble JAM-C prevented melanoma lung metastasis. Together, JAM-C represents a novel therapeutic target for melanoma metastasis.