Effects of Genistein and Daidzein on the Proliferation, Invasion, Migration and Adhesion of Melanoma Cells

IntroductionGenistein and daidzein (Fig.1 ) ,two majorisoflavonoids in soybeans,were reported to playimportant roles in cancer prevention[1 ,2 ] .Previousstudies demonstrated that genistein and daidzeininhibited the growth of leukemia,breast,colonand prostate cancers[38] .Our earlier studies alsodemonstrated that daidzein enhanced the immunefunction in mice[9,1 0 ] . These functions madegenistein and daidzein promising candidates forcancer prevention. However,it is usually themetastatic disse…     

转录靶向性KDR启动子调控双自杀基因治疗肺癌的实验研究

从人肺癌细胞株中克隆KDR基因的启动子(kinasedomainreceptorpromotor,KDRp),构建KDR基因启动子调控的双自杀基因(CDglyTK)真核表达质粒pcDNA3-KDRp-CdglyTK,将其导入ECV304、L9981和NL9980细胞,建立相应的转基因细胞系,并应用不同的前药处理。体内、外实验结果显示:KDR启动子调控的双自杀基因在KDR高表达人肺癌细胞和人脐静脉内皮细胞靶向表达,而在KDR不表达的正常细胞或正常血管内皮细胞中未检测到双自杀基因表达;联合应用5-FC和GCV处理,对转双自杀基因细胞的杀伤作用显著高于单独应用5-FC或GCV,且二者显示了良好的药物协同作用。     

Establishment of the model of vascular endothelial cell membrane chromatography and its preliminary application

A model of vascular endothelial cell membrane chromatography was established by using an ECV304 cell membrane stationary phase (ECV304 CMSP) prepared by immobilizing the ECV304 cell membrane onto the surface of silica carrier. The surface and chromatographic characteristics of ECV304 CMSP were studied. The active component from Caulophyllum robustum was screened by using the model of vascular endothelial cell membrane chromatography. The interaction between the active component and membrane receptor was determined by using a replace experiments. The effect of the active component was tested by using tube formation of ECV304 cell. The results indicated that the model of ECV304 cell membrane chromatograph (ECV304 CMC) can stimulate the interaction between drug and receptor in vitro and the retention characteristics of taspine as active component was similar to that of model molecule in the model of ECV304 CMC. And therefore, taspine acted on VEGFR2 and inhibited the tube formation of ECV304 cell induced by VEGF. This model can be used to screen definite active component as a screening model.     

Effects of Genistein on Cell Cycle and Apoptosis of Two Murine Melanoma Cell Lines

The effects of genistein on several tumor cell lines were investigated to study the effects of gen- istein on cell growth, cell cycle, and apoptosis of two murine melanoma cell lines, B16 and K1735M2. These two closely related murine melanoma cell lines, however, have different responses to the genistein treat- ment. Genistein inhibits the growth of both the B16 and K1735M2 cell lines and arrests the growth at the G2/M phase. After treatment with 60 μmol/L genistein for 72 h, apoptosis and caspase activities were de- tected in B16 cells, while such effects were not found in K1735M2. Further tests showed that after genistein treatment the protein content and mRNA levels of p53 increased in B16, but remained the same in K1735M2. The protein content and mRNA levels of p21WAF1/CIP1 increased in both cell lines after treatment. The results show that genistein might induce apoptosis in B16 cells by damaging the DNA, inhibiting topoi- somerase II, increasing p53 expression, releasing cytochrome c from the mitochondria, and activating the caspases which will lead to apoptosis.     

EVn-50抑制人宫颈癌Hela细胞增殖及诱导凋亡的研究

目的探讨EVn-50对体外培养的人宫颈癌Hela细胞增殖及凋亡的影响。方法体外培养人宫颈癌Hela细胞,台盼蓝拒染法检测EVn-50对人宫颈癌Hela细胞增殖的影响;平皿克隆形成法、软琼脂克隆形成法测定EV-n50对人宫颈癌Hela细胞锚定依赖性生长和锚定非依赖性生长作用的影响;AO／EB染色荧光显微镜观察EVn-50诱导Hela细胞凋亡的形态学改变;DNA凝胶电泳确证EVn-50诱导Hela细胞凋亡作用。结果EVn-50对体外培养人宫颈癌Hela细胞的增殖具有显著抑制作用,呈浓度依赖性。EVn-50可诱导Hela细胞凋亡,AO／EB染色可见典型凋亡小体;DNA凝胶电泳在EVn-50浓度100μg／mL作用48h出现典型“梯形”DNA条带。结论EVn-50具有抑制人宫颈癌Hela细胞增殖并诱导细胞凋亡作用。     

SIOC-AA-005的抗肿瘤作用机制研究

探讨SIOC-AA-005抗肿瘤的作用机制.采用测定HT-29 细胞线粒体膜流动性、线粒体ATP含量、线粒体ATP酶活性及NADH-CoQ氧化还原酶活性、细胞质膜NADH氧化酶活性的方法,对其作用机制进行了深入探讨;SIOC-AA-005在10 nmol/L剂量下可明显降低线粒体膜的流动性,并使线粒体膜电位显著升高;在此浓度下,SIOC-AA-005还可使HT-29细胞内的ATP耗竭,ATP酶活性受到抑制,线粒体中能量产生过程受阻.在胞外反应体系中,SIOC-AA-005对NADH-CoQ氧化还原酶有强烈的抑制作用,对细胞质膜NADH氧化酶也具有较强的抑制作用.SIOC-AA-005可能是通过抑制线粒体NADH-CoQ氧化还原酶、抑制细胞质膜NADH氧化酶以及线粒体ATP酶活性,从而使线粒体膜流动性下降,膜电位升高,细胞内ATP耗竭,最终使肿瘤细胞死亡。     

金雀异黄素对小鼠黑色素瘤B16BL6细胞增殖、迁移和黏附的影响

利用四甲基偶氮唑盐微量酶反应比色法、细胞划痕实验和细胞黏附实验等研究了金雀异黄素对小鼠黑色素瘤B16BL6细胞增殖、迁移以及与血管内皮细胞黏附的影响;利用免疫荧光实验观察了金雀异黄素对B16BL6细胞中微管和肌动蛋白分布的影响.结果表明,与对照组相比,金雀异黄素可以抑制B16BL6细胞的增殖、迁移和与血管内皮细胞的黏附,影响B16BL6细胞中肌动蛋白的组装,但不影响细胞中微管的组装.这些结果提示,金雀异黄素抑制B16BL6细胞的增殖、迁移和与血管内皮细胞的黏附可能是通过诱导细胞中肌动蛋白的重排来实现的.     

七叶皂苷钠对宫颈癌瘤株体内外增殖的抑制作用

为了考察七叶皂苷钠的抗肿瘤活性并探讨其机制,研究了七叶皂苷钠对小鼠宫颈癌U_（14）的体内增殖抑制作用和对人宫颈癌Hela细胞的体外增殖抑制作用．建立昆明种小鼠的U_（14）宫颈癌模型,腹腔注射不同剂量七叶皂苷钠,观察瘤重及胸腺和脾脏指数．用MTT法检测不同浓度七叶皂苷钠对人宫颈癌Hela细胞增殖的影响,AO／EB双染色和流式细胞术检测细胞凋亡,同时考察了七叶皂苷钠对Hela细胞的细胞周期调控作用．结果表明,2．8（mg／kg）·d的七叶皂苷钠可明显抑制小鼠宫颈癌U_（14）的体内增殖（P〈0．01）,七叶皂苷钠对Hela细胞体外生长的抑制率呈时间和浓度依赖关系,细胞周期发生G1／S期阻滞并出现细胞凋亡．七叶皂苷钠能够抑制宫颈癌瘤株的体内外增殖,其可能机制是改变细胞周期分布并诱导细胞凋亡．     

Homoharringtonine induces apoptosis of endothelium and down－regulates VEGF expression of K562 cells

Homoharringtonine (HHT) has currently been used successfully in the treatment of acute and chronic myeloid leukemias and has been shown to induce apoptosis of different types of leukemic cells in vitro. Emerging evidence suggests that angiogenesis may play an important role in hematological malignancies, such as leukemia. However, whether HHT can relieve leukemia by anti-angiogenesis is still unknown. We investigated the anti-angiogenesis potential of HHT with the human umbilical vein endothelial cell line (ECV304) and leukemic cell line (K562) in vitro. Cellular proliferation was determined by MTT assay and apoptosis was analyzed by flow cytometry, The mRNA expression of vascular endothelial growth factor (VEGF) was assessed by RT-PCR and VEGF protein production was detected by Western blot. Inhibition of cell proliferation and induction of apoptosis by HHT were discovered in ECV304 cells, and appeared in a dose- and time-dependent manner, Also, treatment with HHT caused down-regulation of VEGF mRNA expression in K562 cells in similar dose- and time-dependent manner and inhibition of VEGF protein production in K562 cells in response to the enhancing concentration of HHT. The results demonstrated that HHT could also induce apoptosis in endothelium and down-regulate VEGF expression in K562 cells. In conclusion, we believe HHT has anti-angiogenesis potential and speculate that HHT might exert its anti-leukemia effects via reduction of angiogenesis.     

磁场处理对酿酒酵母微观结构及细胞膜流动性的影响及作用机理.

摘要：以磁场为外加物理场,采用恒定磁场及循环磁处理方式对酿酒酵母进行磁处理,研究酿酒酵母微观结构及细胞膜流动性的变化。通过透射电子显微镜,观察不同磁感应强度恒定磁场处理后酿酒酵母的微观结构,发现恒定磁场可以促进酵母细胞的生长,群体中以具有中央大液泡为特征的成熟细胞所占比率增大,同时线粒体呈现适应性膨大、增生。以原子力显微镜观察循环磁处理下酵母细胞的三维结构,发现细胞壁表面出现皱缩、穿孔,胞质外流,证实循环磁处理具有脉冲磁场的特征,可产生瞬间高能作用于细胞,从而产生致死效果。用荧光偏振法对两种处理方式下细胞的膜脂流动性进行测定,发现不同强度恒定磁场处理后,膜脂各向异性下降,流动性增强,0.05T磁感应强度下各向异性值下降3.57%,而循环磁处理后各向异性值比对照提高了8.46%,膜脂流动减慢,证实细胞膜为磁场作用的一个靶点。     

人结肠癌中C-erbB-2,EGFR 的蛋白表达

目的探讨人结肠癌中 Ce-erbB2,EGFR 的蛋白表达.方法应用免疫组织化学方法对人结肠癌细胞系LST14,HT10进行检测.结果2株结肠癌细胞系中均有其表达.结论 CerbB-2,EGFR可能具有协同作用.     

4HPR诱导Hela细胞凋亡

目的：实验结果证明4HPR对Hela细胞具有较强的抑制作用,本文从细胞凋亡角度研究4HPR抑制Hela细胞生长的机理。方法：采用电子显微镜、细胞DNA琼脂糖凝胶电泳、流式细胞仪等方法,分析4HPR诱导Hela细胞凋亡的形态学、生化学及细胞生物学改变特征。结果：电子显微镜下：细胞固缩、核膜扭曲、核染色体聚集成块并靠近核膜;细胞DNA被降解,在1．5％琼脂糖凝胶电泳上出现典型的阶梯状图谱（DNALadder）;流式细胞仪检测出现大量亚二倍体细胞核型峰,二倍体细胞峰减少。结论：上述结果表明,4HPR可诱导Hela细胞凋亡。4HPR抑制Hela细胞生长的机理之一是诱导读细胞凋亡。     

两种槐糖脂抑制人宫颈癌Hela细胞活性比较研究

采用MTT比色法测定分别以豆油和葡萄糖为底物发酵合成的两种槐糖脂抑制人宫颈癌Hela细胞的抑瘤率,对两种槐糖脂的抗肿瘤活性进行了初步研究.研究结果表明,两种槐糖脂对人宫颈癌Hela细胞有一定的抑瘤活性,其中以葡萄糖为底物的槐糖脂较以豆油为底物发酵合成的槐糖脂抑瘤效果明显,这为今后槐糖脂的应用提供关键参数.     

Effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of bone marrow stromal cells and on the adipogenic trans-differentiation of osteoblasts

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), alkaline phosphatase (ALP) activity and oil red O assays were used to examine the effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of primary mouse bone marrow stromal cells (MSCs) and the adipogenic trans-differentiation of primary mouse osteoblasts. The results indicated that daidzein, genistein and glycitein at concentrations from 1×10^-8 mol/L to 1×10^-5 mol/L promoted the proliferation of MSCs and osteoblasts; genistein, daidzein and glycitein promoted osteogenic differentiation and inhibited adipogenic differentiation of MSCs, and inhibited adipocytic transdifferentiation of osteoblasts at appropriate concentrations as 17β-estradiol. It suggests that genistein, daidzein and glycitein regulate a dual differentiational process of MSCs into the osteogenic and adipogenic lineages, and trans-differentiational process of primary osteoblasts into the adipocyte lineages, causing a lineage shift toward osteoblast. Protective effects of them on bone may be mediated by a reversal of adipogenesis which may promote the proliferation, differentiation and mineralization of osteoblasts, and make adipocytes secrete less cytokines which may promote osteoclast formation and activation. In addition, the results also indicated that genistein, daidzein and glycitein may be helpful in preventing the development of steroid induced osteonecrosis.     

OA对人肝细胞HL-7702和肝癌细胞Bel-7402增殖的影响和对细胞凋亡的诱导作用研究

 以人肝细胞HL-7702和肝癌细胞Bel-7402为研究对象,通过MTT法、细胞外部形态观察、Annexin V.FITC&;PI双染法及TUNEL法等方法,研究了腹泻性贝毒的主要组分大田软海绵酸（Okadaic acid, OA）对其增殖的影响及对细胞凋亡的诱导作用。结果表明：OA对人两株细胞的增殖均有显著的抑制作用,24 h IC₅₀分别为65 ng/mL和60 ng/mL,且这种抑制作用呈剂量依赖性。OA导致两株细胞的形态均发生明显变化：细胞变圆,脱壁,细胞膜形成泡状突起,最终形成膜包裹的凋亡小体。Annexin V.FITC和PI双染法及TUNEL法检测均发现：OA能够诱导两株细胞发生不同程度的凋亡,且凋亡细胞的比例与OA的浓度相关。以上结果表明：OA能够通过抑制细胞增殖和诱导细胞凋亡而对人肝细胞和肝癌细胞造成影响,且这种影响的程度与OA的浓度和作用时间密切相关。     

新型植物来源生物碱——唐松草阿原碱对人体肿瘤细胞株的细胞毒性研究

研究从毛莨科唐松草属植物尖叶唐松草根部提取的新型醚键双生物碱－唐松草阿原碱对几种人体肿瘤细胞株的毒性作用。唐松草阿原碱以质量浓度从2．5μg/mL到100μg/mL作用于6种体外培养的肿瘤细胞株及2种正常细胞株,对其中4个肿瘤细胞株种表现了强烈的杀伤作用。IC50分别为：宫颈癌Hela37 μg/mL,肝癌LCC60μg/ml,胃癌MGC65μg/mL,非小细胞肺癌PLA－80182μg/mL,对2个正常细胞株的毒性分别为：肝细胞L－0265μg/mL,小鼠成纤维细胞NIH3T378μg/mL,而对另一株肝癌HepG2及乳腺癌细胞MCF－7无明显作用（IC50＞100μg/mL）。     

仙芦抗癌胶囊抗肿瘤作用及其作用机制研究

通过观察仙芦抗癌胶囊对S180A和H22荷瘤小鼠瘤重和生存时间的影响,观察其体内抗肿瘤作用.采用荧光探针DPH法,观察仙芦抗癌胶囊对S180A和H22荷瘤小鼠红细胞膜流动性的影响.实验结果发现仙芦抗癌胶囊(1g/kg,0 5g/kg,0 25g/kg)对S180A小鼠的瘤体生长有显著的抑制作用.仙芦抗癌胶囊(1g/kg,0 5g/kg)对H22小鼠的生存时间有显著的延长作用.仙芦抗癌胶囊高、中、低剂量(1g/kg,0 5g/kg,0 25g/kg)均能够升高S180小鼠红细胞膜流动性.仙芦抗癌胶囊高、中剂量(1g/kg,0 5g/kg)均能够降低H22小鼠红细胞膜流动性,而低剂量组无明显作用.因此仙芦抗癌胶囊对S180A和H22都有显著的抑制作用,抗肿瘤作用机制与其恢复荷瘤小鼠红细胞膜流动性有关.     

As4S4诱导宫颈癌Hela细胞凋亡及相关分子机制研究

研究AS4S4对宫颈癌Hela细胞生长抑制厦诱导凋亡作用，并探讨可能的分子机制。以不同浓度的AS4S4分不同时间段处理Hela细胞，用四甲基偶氮唑蓝（MTT）法测定细胞增殖抑制率；采用流式细胞术测定细胞凋亡；Western blotting测凋亡相关蛋白Bcl-2，Caspase3表达的变化。结果表明，经AS4S4处理的Hela细胞，增殖受到抑制，作用呈明显的时效和量效关系，细胞凋亡率明显增高并呈浓度依赖性；Western blotting示Bcl-2表达下降，Caspase3表达增加。因此，AS4S4对Hela细胞具有增殖抑制和诱导凋亡作用，其机制与下调Bcl-2蛋白、上调Caspase3蛋白表达有关。     

沙眼衣原体抑制宿主细胞凋亡的研究

目的探讨沙眼衣原体D血清型感染对宿主细胞凋亡的影响。方法沙眼衣原体D血清型感染的Hela229细胞经凋亡诱导剂依托泊苷（etoposide）作用后,Hoechst33．258染色、荧光显微镜观察核浓缩和凋亡小体,流式细胞仪检测凋亡率。结果经依托泊苷作用后,未感染的Hela229细胞有凋亡形态学特征,沙眼衣原体感染的Hela229细胞则无明显凋亡形态学改变,两者的凋亡率比较有统计学意义（P〈0．05）。结论沙眼衣原体感染后能抑制诱导剂诱导的宿主细胞凋亡。     

PPARγ罗格列酮对HT-29的生长抑制作用

PPARγ配体在抗肿瘤中发挥着重要的作用,这些配体具有抗肿瘤、诱导分化和抗血管生成等效果.目的主要是评估PPARγ激动剂罗格列酮对结肠癌细胞株HT-29生长的抑制作用.结果显示,罗格列酮可以有效地抑制体外培养的结肠癌细胞株HT-29的生长及克隆形成,并能抑制HT-29裸鼠移植瘤的生长.与此同时,罗格列酮能够升高PPARγ, p- PPARγ的表达水平.以上结果初步证明罗格列酮在体内外均可抑制结肠癌细胞株HT-29的成长,提示罗格列酮有可能发展成为治疗结肠癌的有效药物.