大豆低聚糖对人胃癌细胞株BGC-823细胞的细胞凋亡和细胞周期的影响

目的通过观察大豆低聚糖对胃癌癌细胞株BGC-823细胞的细胞周期和细胞凋亡的影响,探索乳酸杆菌发酵滤液对胃癌细胞作用的可能机制。方法用光镜和流式细胞仪分析不同浓度大豆低聚糖对BGC-823细胞的凋亡诱导效果;用流式细胞仪分析不同浓度大豆低聚糖对BGC-823细胞细胞周期的影响。结果大豆低聚糖可以诱导BGC-823细胞的凋亡。形态学观察处理后的BGC-823细胞,可见细胞变形,细胞皱缩,体积变小,细胞间隙增大,细胞核固缩。流式细胞仪分析50 mg/ml和100 mg/ml大豆低聚糖作用48 h和72 h BGC-823细胞的凋亡比例,分别为6.76%和7.93%。50 mg/ml大豆低聚糖作用48 h,引起BGC-823细胞G1期阻滞,100 mg/ml大豆低聚糖作用48 h,引起BGC-823细胞出现S期阻滞。结论大豆低聚糖可诱导部分BGC-823细胞凋亡。大豆低聚糖对BGC-823细胞的生长抑制作用在低浓度时可能通过G1期阻滞实现,在高浓度时可能通过S期阻滞实现。     

无花果枝提取物体外诱导胃癌BGC-823细胞凋亡的研究

为探讨新鲜无花果枝提取物(FBE)对体外培养的人胃癌细胞株BGC-823增殖和凋亡的影响,用不同浓度FBE处理胃癌BGC-823细胞,采用细胞形态学观察,细胞活力测定(MTY法)及免疫组织化学法检测增殖细胞核抗原(PCNA)表达情况来评价FBE对BGC-823细胞体外增殖的影响,应用流式细胞术的膜联蛋白V/碘化丙啶技术(Annexin V/PI法)检测FBE诱导BGC-823细胞体外凋亡的情况.形态学观察发现中高剂量(＞0.5mg/mL)组处理4 h细胞出现明显凋亡,24 h检测到中高剂量(＞0.5 mg/mE)组细胞增殖活力和增殖细胞核抗原表达显著降低(P＜0.05),流式细胞仪检测到FBE处理4 h所有剂量组细胞平均凋亡率(9.76%,10.87%,14.29%,49.67%,71.37%)均高于对照组(6.1%)(P＜0.05或P＜0.01),以上作用效果呈现剂量依赖性.以上结果说明无花果枝提取物能够通过诱导胃癌BGC-823细胞凋亡从而抑制其体外的生长与增殖.     

蛹虫草菌丝生长研究

通过对 4种不同来源的蛹虫草菌种 ,在 5种不同配方培养基上菌丝生长情况进行了比较研究。结果表明 ,不同培养基配方对不同来源的蛹虫草菌种培养菌丝的长速和长势存在较大的差异。     

真菌激发子对提高蛹虫草虫草菌素的作用*

研究了不同真菌激发子菌株对提高蛹虫草生物量和虫草菌素含量的影响,其中一株疫霉(Phytophthora sp.)YL提高虫草菌素含量比对照高4倍。机械研磨和80℃高温细胞自溶相结合制备的真菌激发子诱导效果最佳。同一激发子不同浓度对虫草素的诱导实验表明, 80mg/ml浓度碳水化合物的真菌激发子诱导蛹虫草产生虫草菌素效果最明显。     

稀土化合物氯化亚鈰对人肺癌细胞PG、人胃癌细胞BGC-823的作用

 以人肺癌细胞 Ｐ Ｇ 和人胃癌细胞 Ｂ Ｇ Ｃ ８２３ 作为研究对象,利用 Ｍ Ｔ Ｔ 测定、３ Ｈ Ｔｄ Ｒ 参入、流式细胞术、软琼脂培养、 Ｎｏｒｔｈｅｒｎ ｂｌｏｔ、 Ｗ ｓｔｅｒｎ ｂｌｏｔ 等实验方法,观察了稀土化合物氯化亚鈰（ Ｃｅ Ｃｌ３）抑癌作用．结果表明, Ｃｅ Ｃｌ３ 浓度为 ００５ ｍ ｍ ｏｌ／ Ｌ,０１ ｍ ｍ ｏｌ／ Ｌ,０５ ｍ ｍ ｏｌ／ Ｌ和 １ ｍ ｍ ｏｌ／ Ｌ可抑制 Ｐ Ｇ 细胞的增殖;浓度为 ０５ ｍ ｍ ｏｌ／ Ｌ和 １ ｍ ｍ ｏｌ／ Ｌ可抑制 Ｐ Ｇ 细胞 Ｄ Ｎ Ａ 的合成,其 Ｇ１ 期细胞比例增加而 Ｓ期细胞比例减少,在软琼脂中的生长能力降低,原癌基因 ｃ ｍ ｙｃ 和 ｃ ｒａｓ 表达降低,ｐ１６ 蛋白质表达降低．而同样浓度的 Ｃｅ Ｃｌ３ 对 Ｂ Ｇ Ｃ ８２３ 细胞和正常细胞 ２ Ｂ Ｓ未见影响．提示：稀土化合物抑制肺癌细胞 Ｐ Ｇ 的增殖以及降低其恶性度的作用机制可能与一些增殖相关的原癌基因的表达和细胞周期的调控有关,其确切的机理还需进一步的研究．     

靶向survivin基因的siRNA对姜黄素诱导胃癌细胞凋亡的影响

目的:研究靶向survivin基因的siRNA对胃癌细胞,survivin表达的影响,抑制survivin基因表达对姜黄素诱导胃癌细胞凋亡的影响。方法:通过脂质体将survivinsiRNA导入胃癌细胞株BGC-803,用Real-timePCR和Western-blotting检测转染后细胞内survivin基因表达水平,流式细胞仪和Hochest染色检测细胞凋亡的改变。结果:姜黄素可抑制BGC-803细胞的生长,其生长抑制率和药物浓度与作用时间呈依赖关系;姜黄素作用BGC-803细胞后,survivin蛋白和mRNA表达降低;通过转染survivinsiRNA抑制BGC-803细胞survivin基因的表达能促进姜黄素诱导BGC-803细胞凋亡的作用。结论:靶向抑制survivin基因表达后姜黄素诱导胃癌细胞BGC-803凋亡的作用增强。     

姜黄素对人食管癌EC9706细胞凋亡的诱导作用

目的:应用姜黄素处理人食管癌EC9706细胞,研究姜黄素对人食管癌EC9706细胞凋亡的诱导作用。方法:应用细胞计数、流式细胞仪、琼脂糖凝胶电泳、Hoechst染色、H.E染色和透射电镜检测经姜黄素诱导处理后人食管癌EC9706细胞的凋亡。结果:经姜黄素诱导处理后,人食管癌EC9706细胞生长抑制率达69.9%;细胞周期检测出现亚二倍体(亚G1期)细胞峰值,细胞凋亡率达23%;琼脂糖凝胶电泳显示出细胞凋亡典型的180-200 bp及其倍体的DNA"梯状"条带;Hoechst染色显示细胞核内出现浓染致密的固缩形态或颗粒状荧光;光镜和电镜下可见典型的细胞凋亡特征:细胞体积缩小,染色体凝集,可见有成群或单独存在的凋亡细胞,电镜下可见凋亡小体存在。结论:姜黄素能够有效诱导人食管癌EC9706细胞的凋亡,从而进一步为食管癌等恶性肿瘤疾病的治疗和凋亡机理的研究提供重要基础和科学依据.     

红车轴草提取物对胃癌BGC-823细胞凋亡的影响

探讨红车轴草(TrifoliumpratenseL)提取物对胃癌细胞株BGC-823的抑制增殖效应及诱导凋亡作用.我们采用不同浓度(50、100、250、500、1000mg/L)红车轴草提取物处理BGC-823细胞,采用四甲基偶氮唑盐(MTT)法检测药物对细胞的抑制作用,倒置显微镜观察细胞的形态学改变;AO/EB染色,荧光显微镜观察细胞凋亡形态;采用DNALadder观察DNA的降解;应用流式细胞仪观察细胞凋亡过程中的细胞周期的变化和细胞凋亡.结果显示在红车轴草提取物的作用下,BGC-823细胞呈凋亡改变,DNA琼脂糖凝胶电泳呈典型的凋亡特征.细胞凋亡的同时,细胞周期阻滞于G2/M期.实验结果表明红车轴草提取物能抑制胃癌BGC-823细胞增殖,能诱导胃癌细胞凋亡.     

蛹虫草对腺嘌呤致肾阳虚证小鼠影响的实验研究

目的研究人工栽培的蛹虫草对腺嘌呤致肾阳虚小鼠的防治作用。方法将小鼠随机分为正常对照组,肾阳虚模型组和蛹虫草低、中、高剂量组(1.25 g·kg-1、2.5 g·kg-1、5 g·kg-1),除正常对照组外,各组均用腺嘌呤(25 mg·kg-1)造模,给药组同时灌胃给予3个剂量的蛹虫草子实体细粉悬液,连续处理28 d。实验结束后,测定小鼠体质量、自主活动次数、低温游泳时间、脏器湿重、血清睾酮和皮质醇含量,并同时观察小鼠睾丸组织病理结构变化。结果与正常对照组小鼠比较,肾阳虚模型组小鼠体质量明显下降,自主活动次数减少,低温游泳时间变短,血清睾酮和皮质醇的含量降低,睾丸组织发生了退行性变化。与肾阳虚模型组小鼠相比,给予适当剂量的蛹虫草后,小鼠的体质量增加,自主活动次数增多,低温游泳时间延长,脏器湿重增加,血清睾酮和皮质醇含量显著升高,睾丸组织损伤减轻。结论蛹虫草对腺嘌呤致肾阳虚小鼠有一定的防治作用。     

靶向survivin的siRNA对胃癌BGC-823细胞增殖的影响

目的探讨干扰RNA沉默生存素（survivin）基因表达对人胃癌BGC-823细胞增殖和凋亡的影响。方法设计并合成3条靶向survivin的小分子干扰RNA（siRNA）,构建表达性干扰RNA质粒（shRNA）——shRNA-survivin-1、shRNA-survivin-2和shRNA-survivin-3,分别转染胃癌BGC-823细胞,实时定量PCR检测干扰RNA沉默survivin mRNA表达效果,Westernblot观察对胃癌BGC-823细胞survivin蛋白质表达的抑制,MTT（四甲基偶氮唑盐）比色法分析检测细胞生长抑制率,流式细胞计数检测各组细胞周期和凋亡率,探讨干扰RNA对胃癌BGC-823细胞生长的影响。结果在体外,shRNA-survivin-1有效沉默人胃癌BGC-823细胞survivin mRNA的表达,使sur-vivin mRNA相对水平明显降低（P〈0.05）,survivin蛋白质表达抑制,72h细胞生长抑制率达74.92%（P〈0.05）,shRNA-survivin-1使G2/M期细胞百分比明显增加,凋亡率显著增加（P〈0.05）。结论 shRNA-survivin-1可以沉默survivin基因的表达,可以显著抑制胃癌BGC-823细胞的增殖,在一定程度上诱导其自发凋亡。本研究为靶向sur-vivin的RNA干扰在胃癌的基因治疗提供了有力的理论依据和技术储备。     

靶向survivin的siRNA对胃癌BGC-823细胞增殖和转移能力的影响

目的探讨沉默生存素（survivin）基因表达的干扰RNA对人胃癌BGC-823细胞增殖和成瘤能力的影响。方法应用已经在细胞上验证能够有效沉默survivin的小分子干扰RNA（shRNA-survivin-1）,并在体外实验的基础上,建立稳定表达干扰RNA细胞系,进一步探讨干扰RNA稳定表达对胃癌BGC-823细胞生长和裸鼠移植成瘤的影响。结果 shRNA-survivin-1有效沉默人胃癌BGC-823细胞survivin mRNA的表达,成功筛选shRNA-sur-vivin-1稳定表达细胞株BGC/siRNA-1细胞,实验表明,BGC/siRNA-1细胞的生长曲线缓慢上升,细胞增殖能力下降;BGC/siRNA-1细胞裸鼠移植成瘤体积与对照组相比,明显减小（P〈0.05）。结论 shRNA-survivin-1可以沉默survivin基因的表达,可以显著抑制胃癌BGC-823细胞的增殖,并降低胃癌BGC-823细胞的成瘤能力,本研究为靶向survivin的RNA干扰在胃癌的基因治疗提供了有力的理论依据和技术储备。 更多还原     

Inhibition of cell proliferation and induction of apoptosis by ExFABP gene targeting

Ex-FABP, an extracellular fatty acid binding lipocalin, is physiologically expressed by differentiating chicken chondrocytes and myoblasts. Its expression is enhanced after cell treatment with inflammatory stimuli and repressed by anti-inflammatory agents, behaving as an acute phase protein. Chicken liver fragments in culture show enhanced protein expression after bacterial endotoxin treatment. To investigate the biological role of Ex-FABP, we stably transfected proliferating chondrocytes with an expression vector carrying antisense oriented Ex-FABP cDNA. We observed a dramatic loss of cell viability and a strong inhibition of cell proliferation and differentiation. When chondrocytes were transfected with the antisense oriented Ex-FABP cDNA we observed that Ex-FABP down-modulation increased apoptotic cell number. Myoblasts transfected with the same expression vector showed extensive cell death and impaired myotube formation. We suggest that Ex-FABP acts as a constitutive survival protein and that its expression and activation are fundamental to protect chondrocytes from cell death.     

Retracted: Curcumin derivative L6H4 inhibits proliferation and invasion of gastric cancer cell line BGC-823

Curcumin and its chalcone derivatives have well-known, explicit biological antitumor properties, such as instance antiproliferative and apoptotic effects via multiple molecular targets. In this study, we investigated the anticancer activity of curcumin derivative L6H4 (curcumin L6H4) on gastric cancer cells. Inhibitory effects of curcumin L6H4 on gastric cancer cells (BGC-823) were studied by the diphenyltetrazolium (MTT) assay, and cell apoptosis was detected by Annexin-V/propidium iodide (PI) staining and then analyzed by flow cytometry. A mouse xenotransplant gastric tumor model was established to detect the role of curcumin L6H4 in vivo. The apoptosis-related proteins p53, p21, Bax, and Bcl-2 in BGC-823 cells and mouse xenotransplant models treated with curcumin L6H4 were determined by Western blot analysis. Curcumin L6H4 can significantly inhibit the proliferation and induce the apoptosis of BGC-823 cells, thus enhancing the expression levels of p53, p21, Bax, and Bcl-2 noticeably in vivo and in vitro. Meanwhile, curcumin L6H4 can remarkably suppress the growth of tumor cells in animal models. These results suggest that curcumin derivative L6H4 has potent of antitumor properties in vitro or in vivo.     

Inhibition of cell proliferation and induction of apoptosis by genistein in experimental hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the leading cause of cancer related deaths in the world, with increasing incidence in many developed countries. Epidemiological data suggest that consumption of soy products may be associated with a decreased risk of cancer. We investigate the effects of genistein on cell proliferation, apoptosis and caspase-3 in DEN induced (200 mg/kg body weight; by single intraperitoneal injection) and Phenobarbital promoted (0.05% through drinking water for 14 successive weeks) cancer-bearing rats. Immunohistochemistry was employed to detect cell proliferating markers proliferating cell nuclear antigen (PCNA), DNA fragmentation was determined by agarose gel electrophoresis and terminal deoxynucleatide transferase dUTP nick labeling (TUNEL) staining and caspase by enzyme-linked immunosorbent assay. We found inhibition of cell proliferation, induction of apoptosis and activation of caspase-3 in genistein treated animals. From these results, we conclude that genistein inhibit cell proliferation, induced apoptosis. This activation of caspsase-3 in genistein treated liver cancer bearing animals correlated well with its apoptosis inducing effect.     

人胃癌BGC-823细胞中去甲斑蝥素抑癌作用机理的蛋白质组学研究

去甲斑蝥素是我国自行研制的抗肿瘤药物,在临床上主要用于消化道肿瘤的治疗．实验表明,去甲斑蝥素可引起人胃癌BGC-823细胞发生 M期阻滞及细胞凋亡．进一步利用双向电泳和质谱技术,筛选出了去甲斑蝥素抑癌作用相关蛋白．研究显示,线粒体热休克蛋白CH60、线粒体ATP合酶d亚单位、内质网葡萄糖调节蛋白GRP78、线粒体Hsp70的辅助因子GRPE1、SH3L3以及染色质组装因子1小亚基RBBP4参与了去甲斑蝥素的抑癌作用．研究提示,去甲斑蝥素可能通过促进线粒体热休克蛋白及p53的表达进而激活caspase-3依赖的凋亡通路,并且去甲斑蝥素在引发内质网协迫之后,可通过抑制胞外信号调节激酶(extracellular signal regulated kinase, ERK)的活性促进肿瘤细胞的凋亡．进一步分析了去甲斑蝥素与线粒体ATP合酶抑制剂寡霉素A的联合用药对人胃癌细胞生长的影响,结果表明,联合用药的抑瘤效果比单独用药的抑瘤效果显著,提示去甲斑蝥素可能通过抑制线粒体ATP合酶功能抑制BGC-823生长．上述结果为优化去甲斑蝥素的联合用药方案提供了新线索．     

Dihydroartemisinin induces apoptosis in human gastric cancer cell line BGC-823 through activation of JNK1/2 and p38 MAPK signaling pathways

Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, is associated with a broad range of biological properties including antitumor activity. However, the effect of DHA on gastric cancer has not been clearly clarified. The aim of this study was to investigate the role and mechanism of DHA in human gastric cancer cell line BGC-823. Cell viability was assessed by MTT assay. Cell apoptosis was analyzed with flow cytometry. The expressions of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38 MAPK) and their phosphorylated forms as well as apoptosis related proteins were examined by western blot analysis. The results demonstrated that DHA inhibited cell viability of BGC-823 cells in a dose- and time-dependent manner. DHA treatment upregulated the expression of Bax, cleaved caspase-3 and -9, and degraded form of PARP, and downregulated the Bcl-2 expression and Bcl-2/Bax ratio. Meanwhile, DHA increased the phosphorylation of ERK1/2, JNK1/2 and p38 MAPK. Synthetic inhibitors of JNK1/2 or p38 MAPK kinase activity, but not inhibitor of ERK1/2, significantly abolished the DHA-induced activation of caspase-3 and -9. In vivo tumor-suppression assay further indicated that DHA displayed significant inhibitory effect on BGC-823 xenografts in tumor growth. These results indicate that DHA induces apoptosis of BGC-823 cells through JNK1/2 and p38 MAPK signaling pathways and DHA could serve as a potential additional chemotherapeutic agent for treatment of gastric cancer.     

Octa-substituted anionic porphyrins: topoisomerase I inhibition and tumor cell apoptosis induction

Beta-octabromo- meso-tetra(4-carboxyl)phenyl porphyrin 6 and beta-octaphenyl- meso-tetra(4-carboxyl)phenyl porphyrin 8 were synthesized and fully characterized by (1)H NMR, UV, and HRMS. Their cytotoxicities to tumor cells were tested using MTT assays. One kind of tumor cell apoptosis induced by these anionic porphyrins under irradiation was examined by flow cytometric analysis. The inhibition of Topo I (Topoisomerase I) indicates that Topo I preferentially binds to the synthesized compounds, thus blocking the interaction between Topo I and DNA. The results implied that compounds 4, 6, and 8 are potential inhibitors to Topo I, which might be one of the important factors inducing apoptosis of tumor cells.