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。     

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对小鼠肝癌H22腹水瘤的治疗研究

目的：研究人参皂甙Rg3治疗小鼠恶性腹腔积液的疗效。方法：100只雌、雄各半昆明种小鼠随机分为5组：Ⅰ组生理盐水组（0．9％NS）;Ⅱ组顺铂组（DDP0．5mg／kg）;Ⅲ组低剂量人参皂甙Rg3组（LPD0．3mg／kg）;Ⅳ组中剂量人参皂甙Rg3组（MPD1．0mg／kg）;Ⅴ组高剂量人参皂甙Rg3组（HPD3．0mg／kg）。所有小鼠肝癌H22腹水瘤模型建立后24小时开始分别腹腔注射0．2ml／只治疗,每日1次,共14天,同时观察小鼠的生活状态和体重。治疗结束后24小时,处死各组半数小鼠测各项指标,剩余小鼠停止治疗并观察生存时间。结果：各用药组较生理盐水组在腹水量（P〈0．05）、腹水中瘤细胞数和瘤细胞存活率方面都下降（P〈0．05）;随着人参皂甙Rg3给药浓度的增加,腹水中瘤细胞数及瘤细胞存活率下降（P〈0．05）;人参皂甙Rg3高剂量组腹水中瘤细胞数及瘤细胞存活率低于DDP组（P〈0．05）。各实验用药组较生理盐水组小鼠寿命延长,其中中剂量人参皂甙Rg3组延长最明显（P〈0．05）。结论：人参皂甙Rg3对小鼠恶性腹腔积液的形成具有抑制作用,这为临床应用提供了实验依据。     

槲皮素联合顺铂对肺腺癌小鼠移植瘤Bcl-2和Bax表达的调控及意义

目的：观察槲皮素联合顺铂对肺腺癌LA795细胞的T739小鼠移植瘤的抑制作用，检测移植瘤中Bcl-2和Bax的表达水平及肿瘤细胞凋亡指数，探讨其作用机制。方法：将32只接种了LA795肺腺癌细胞的T739小鼠随机分成对照组（A组）、顺铂组（B组）、槲皮素组（C组）、槲皮素＋顺铂组（D组）。用药16d后，观察肿瘤生长情况，接种后24d活杀各组小鼠，移植瘤标本称瘤体质量和测量体积，免疫组织化学和图像分析系统定量检测肿瘤组织的Bcl-2和Bax表达，原位凋亡TUNEL法检测肿瘤细胞凋亡指数。结果：B、C、D各用药组肿瘤的生长受到明显抑制，瘤体质量明显低于A组，其抑瘤率分别为38．57％、26．03％和51．58％，联合用药组抗瘤作用进一步增强。Bcl-2表达在A组高于B、C、D组，Bax表达在A组明显低于B、C、D组，各用药组与对照组相比差异均有统计学意义（P〈0．05或P〈0．01）；肿瘤细胞的凋亡指数在各用药组都比对照组明显提高，差异有统计学意义（P〈0．01）。结论：槲皮素联合顺铂可明显抑制肺腺癌细胞在小鼠体内的生长，其作用机制可能是通过调控肿瘤中Bcl-2和Bax的表达，从而抑制细胞增殖和促进细胞凋亡。     

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.     

人参皂苷Rg3与顺铂对荷卵巢癌裸鼠免疫功能及肿瘤相关蛋白的影响

[目的]观察人参皂苷Rg3联合顺铂对荷高转移人卵巢癌细胞系HO．8910PM转移瘤裸鼠白细胞介素．2（IL-2）、干扰素-γ（INF-γ）及nm23、血管内皮生长因子（VEGF）及增殖细胞核抗原（PCNA）的影响。[方法]移植瘤裸鼠随机分成6组腹腔给药：荷瘤空白对照组fA组）;不同Rg3浓度药物组：2．5mg／kg（B组）、5mg／kg（C组）和lOmg／kg（D组）;Rg3和顺铂联用组（E组）以及顺铂阳性对照组（F组）。分别测定各组裸鼠转移瘤体积变化、IL-2、INF-γ、nm23、VEGF及PCNA的变化情况。[结果]与A组比较,E组（P〈0．05）和F组（P〈0．01）小鼠血清中IL-2、INF-γ含量均能显著降低;B（P〈0．05）、C、D和E（P〈0．01）组小鼠移植瘤组织nm23的表达均增强。Rg3给药后,小鼠移植瘤组织VEGF、PCNA表达均呈下降趋势。[结论]人参皂苷Rg3和顺铂联用具有协同抗肿瘤及减毒的作用。Rg3可能通过增强mm23的表达．使VEGF及PCNA表达呈下降趋势等多途径产生抗肿瘤作用。     

苦参碱对小鼠H22肝癌细胞凋亡作用的实验研究

目的：研究中药苦参碱在体内外对小鼠H22肝癌细胞的诱导凋亡作用，探讨其可能的抗肿瘤作用机制。方法：Annexin V-FITC／PI双标记法检测苦参碱对H22细胞的早期促凋亡作用；免疫组织化学法检测苦参碱作用后H22细胞内Bcl-2、Bax两种凋亡相关蛋白的表达。建立H22肝癌移植瘤小鼠模型，观察苦参碱对荷瘤小鼠肿瘤生长情况的影响及肿瘤抑制率；电镜观察荷瘤小鼠肿瘤组织的超微结构改变；免疫组化方法检测小鼠肿瘤组织内Bcl-2、Bax的表达情况。结果：AnnexinV法检测到1．0mg／mL和1．5mg／mL苦参碱作用48h后H22细胞有早期凋亡改变，凋亡细胞百分率分别为11．71％和17．86％，与对照组相比差异有统计学意义（P〈0．05）。苦参碱对荷瘤小鼠肿瘤的抑制率达60％以上；免疫组化显示，苦参碱作用后H22细胞内及小鼠肿瘤组织内Bcl-2蛋白的表达水平下降，Bax蛋白表达增强。电镜证实苦参碱治疗后荷瘤小鼠肿瘤组织内有凋亡细胞和凋亡小体的存在。结论：苦参碱在体内体外都对小鼠H22肝癌细胞表现出较强的抗肿瘤作用和诱导凋亡作用。促凋亡作用与其上调细胞内Bax表达，抑制Bcl-2表达有关。     

低氧条件下人参皂甙Rg3诱导人喉鳞癌细胞凋亡及对HIF-1α和VEGF表达的影响

 目的研究人参皂甙Rg3对低氧条件下人喉鳞癌细胞(Hep-2)生长抑制作用以及可能的机制。方法通过体外低氧培养Hep-2人喉鳞癌细胞株,同时设立阴性对照组,阳性对照组(顺铂);采用TUNEL法(终末脱氧核苷酸末端转移酶介导缺口末端标记)检测细胞凋亡情况,计数凋亡细胞,计算凋亡率;流式细胞术测定细胞周期及细胞凋亡情况;应用免疫细胞化学技术和流式细胞技术检测Hep-2细胞在人参皂甙Rg3作用下HIF-1α和VEGF蛋白表达的变化;应用RT-PCR技术检测HIF-1α和VEGF mRNA表达的变化。结果TUNEL及流式细胞术检测Rg3组的凋亡率明显高于阴性对照组(P<0.05);在低氧条件下,Rg3组和顺铂对照组的Hep-2细胞中的HIF-1α和VEGF蛋白表达低于阴性对照组(P<0.05);Rg3组的HIF-1α和VEGF mRNA水平明显低于阴性对照组(P<0.05),顺铂组的HIF-1α和VEGF mRNA水平无明显变化。结论低氧条件下,人参皂甙Rg3诱导细胞凋亡,抑制了Hep-2细胞中的HIF-1α和VEGF蛋白和mRNA的表达,这可能是Rg3抗肿瘤作用的机制之一。     

抗癌防转汤抑制肝癌淋巴道转移及其对MMP-9和CTL影响的研究

目的：探讨抗癌防转汤抑制小鼠肝癌淋巴道转移效果及其机制。方法：采用随机数字表法将小鼠分为正常对照组、荷瘤对照组、低剂量治疗组和高剂量治疗组，建立小鼠肝癌淋巴道转移模型，计算接种后面（21d小鼠抑瘤率）并运用MTT法检测CTL细胞活性及免疫组织化学方法和Western blot检测各组原发瘤及转移瘤基质金属蛋白酶-9（matrix metalloproteinase9，MMP-9）的表达水平，运用统计学分析数据意义。结果：治疗组较对照组，小鼠抑瘤率明显提高，MMP-9在原发瘤、转移瘤表达均减少（P＜0．05），细胞毒性T淋巴细胞（cyto-toxicity T lymphocyte，CTL）活性增强（P〈0．05），而且跟药物浓度呈正相关，P〈0．05。结论：抗癌防转汤有抑制肝癌淋巴道转移的作用，且其与MMP-9的表达水平降低，CTL细胞活性增强有关。     

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.     

Ginsenoside Rg3 attenuates tumor angiogenesis via inhibiting bioactivities of endothelial progenitor cells

Accumulating evidence suggests that Ginsenoside Rg3 appears to inhibit tumor growth including Lewis lung carcinoma, intestinal adenocarcinomas or B16 melanoma by inhibiting cell proliferation, tumor cell invasion and metastasis. Endothelial progenitor cells (EPCs) appear to play a key role in the growth of early tumors by intervening with the angiogenic switch promoting tumor neovessel formation by producing angiogenic cytokines during tumor progression. This paper reports a novel mechanism of Ginsenoside Rg3, a candidate anticancer bio-molecule, on tumor angiogenesis by inhibiting the multiple bioactivities of EPCs. When Ginsenoside Rg3 was applied to the ex vivo cultured outgrowth ECs, a type of EPCs, it inhibited the cell proliferation, cell migration and tubular formation of EPCs. Importantly, Ginsenoside Rg3 attenuated the phosphorylation cascade of the VEGF dependent p38/ERK signaling in vitro. The xenograft tumor model clearly showed that Ginsenoside Rg3 suppresses tumor growth and tumor angiogenesis by inhibiting the mobilization of EPCs from the bone marrow microenvironment to the peripheral circulation and modulates VEGF-dependent tumor angiogenesis. In conclusion, this study provides a potential therapeutic molecule, Ginsenoside Rg3, as an anticancer drug by inhibiting the EPC bioactivities.     

Effects of total-body hyperthermia on metastases from experimental mouse tumors

To study the effects of total-body hyperthermia (TBH) on metastases from malignant tumors, Lewis lung carcinoma (LLC)-bearing C57BL/6 mice and mouse ascites hepatoma 134-bearing C3H/He mice were immersed in a heated water bath. Rectal temperature was maintained for 30 min at 40 degrees C or 42 degrees C. After treatment, the incidence of lung metastasis was analyzed in LLC-inoculated mice, and the presence or absence of metastasis in affiliated lymph nodes was determined in mouse ascites hepatoma 134-inoculated mice. A significant inhibition in primary tumor growth in LLC- and mouse ascites hepatoma 134-bearing mice treated with 42 degrees C TBH was noted. The incidence of lung metastasis was increased from the control level of 1.6 +/- 0.63 (SD) to 2.4 +/- 0.98 in the 42 degrees C TBH (P less than 0.01) groups but not in the 40 degrees C TBH group. Metastasis to affiliated lymph nodes was similar for the controls and the 40 degrees C and 42 degrees C TBH groups. The increase in lung metastasis in LLC-treated mice subjected to 42 degrees C TBH could be prevented by the combined use of anticancer drugs such as cis-diamminedichloroplatinum(II) (1.0, 3.0 mg/kg) or mitomycin C (0.3, 1.0 mg/kg). Furthermore, the combined use of 42 degrees C TBH and anticancer drugs showed the inhibition of primary tumor growth to a greater degree than did 42 degrees C TBH alone or anticancer drugs alone. Since 42 degrees C TBH may induce tumor metastasis, especially hematogenous metastasis, it seems advisable to use anticancer drugs in combination with clinical thermal applications.     

顺铂联合人参皂甙Rg3对荷宫颈癌鼠血管生成的抑制作用

目的探讨人参皂甙Rg3(简称Rg3)与顺铂联合应用对人宫颈癌动物模型中肿瘤的生长及其对肿瘤血管生成的影响。方法采用人宫颈癌HeLa细胞株接种于28只雌性裸鼠,随机均分成4组：即（1）生理盐水对照组;（2）顺铂（DDP）组;（3）人参皂甙Rg3组;（4）人参皂甙Rg3与顺铂联合组。各组用药时间均为5周,35天后脱颈处死。分别检测肿瘤的重量,并计数肿瘤内微血管密度(MVD)。结果Rg3组、DDP组及联合组对原位种植肿瘤的生长有明显的抑制作用,联合治疗组抑制肿瘤生长的作用明显优于单纯组;且Rg3及联合治疗组MVD也明显低于DDP组及对照组。结论Rg3与DDP联合应用能抑制人移植性宫颈癌的生长,降低肿瘤内的MVD,抑制宫颈癌肿瘤血管生成。     

人参皂苷Rg3对人鼻咽癌HNE-1细胞增殖和血管生成的影响

 目的 研究人参皂苷Rg3（ginsenoside Rg3）对人鼻咽低分化鳞癌HNE-1细胞的增殖、迁移及体外血管生成的影响。方法 用不同浓度人参皂苷Rg3处理HNE-1细胞,MTT法检测HNE-1细胞增殖活性;创伤修复实验检测细胞迁移能力;观察HNE-1细胞能否在Matrigel上形成血管网状结构及其特点;体外管道形成抑制试验检测不同浓度人参皂苷Rg3对HNE-1细胞管道形成能力的影响。结果 不同浓度人参皂苷Rg3（50、100、200μg/ml）对HNE-1、CRL-2480细胞均有一定的增殖抑制作用,呈浓度依赖性趋势,无时间依赖性,差异均无统计学意义（P>0.05）;人参皂苷Rg3（25、50、100、200μg/ml）可以显著降低HNE-1细胞迁移速度,与对照组比较差异有统计学意义（P<0.001）,与浓度呈负相关(r=-0.964;P<0.001);HNE-1细胞在Matrigel上培养能形成血管网状样结构;人参皂苷Rg3能抑制HNE 1 细胞体外管道形成（P<0.01）,其管状结构数量与人参皂苷Rg3浓度呈负相关（r=-0.928;P<0.01）。结论 人参皂苷Rg3有一定的抗HNE-1细胞增殖作用;HNE 1细胞具有血管生成拟态;人参皂苷Rg3能够抑制HNE-1细胞的迁移和体外血管生成拟态的形成。     

Suppression of VEGFR-3 signaling inhibits lymph node metastasis in gastric cancer

In gastric cancer, lymph node metastasis is one of the major prognostic factors and forms the basis for surgical removal of local lymph nodes. Recently, several studies have demonstrated that overexpression of lymphangiogenic growth factor VEGF-C or VEGF-D induces tumor lymphangiogenesis and promotes lymphatic metastasis in mouse tumor models. We examined whether these processes could be inhibited in naturally metastatic tumors by blocking of their cognate receptor VEGFR-3 signaling pathway. Using a mouse orthotopic gastric cancer model which has a high frequency of lymph node metastasis, we estimated lymphatic vessels in gastric cancers by immunostaining for VEGFR-3 and other specific lymphatic markers, LYVE-1 and prox-1. Then we systemically administered anti-VEGFR-3 blocking antibodies. This treatment resulted in the inhibition of regional lymph node metastasis and reduction of lymphatic vessel density in the primary tumors. In addition, increased density of LYVE-1-positive lymphatic vessels of primary tumors was closely correlated with lymph node metastasis in human samples of gastric cancer. Antilymphangiogenesis by inhibiting VEGFR-3 signaling could provide a potential strategy for the prevention of lymph node metastasis in gastric cancer.