运用产气袋法制造微缺氧培养人胃癌细胞株SGC-7901

目的：运用GasPaK产气袋法制造微缺氧条件培养人胃癌细胞株SGC-7901并观察该细胞在微缺氧条件下超微结构的变化。方法：运用产气袋法(GasPaK)制造微缺氧条件，血气分析监测1640培养液中PO2、PCO2和pH值变化。透射电镜观察微缺氧对SGC-7901细胞超微结构的改变。结果：(1）GasPaK产气袋法在0.5-48 h内PO2、PCO2和pH稳定，并能达到微缺氧细胞培养的条件；(2）微缺氧导致部分SGC-7901细胞超微结构发生改变。结论：通过GasPaK产气袋法制造的微缺氧肿瘤细胞培养环境，具有条件稳定、重复性好的持点。     

The effect of Fufangkushen on gastric cancer cell killing by lines SGC-7901 human γδT cell

Objective To explore the effect of Fufangkushen on gastric cancer cell killing by human γδT cells. Methods Isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells in vitro. Fufangkushen at various concentrations was used to induce γδT cells and gastric cancer cell lines SGC-7901 for 24 hours, MTr assays was used to detect inhibitory effect of Fufangkushen on these cell lines, LDH assays was used to measure the cytotoxic activity of γδT cells, and flow cytometry was used to detect apoptosis of γδT cells and SGC-7901 before and after the treatment. Results Ten days after cultivation, proliferation ra-tio of γδT cells increased from 4.21% to 70.35% and CD44 was up to 94.0%. Inhibitory rate of Fufangkush-en on SGC-7901 at various concentrations was significantly higher than that on γδT cells (22.3% vs-22.4%, P<0.05). The negative inhibitory ratio on γδT cells showed a dose-dependent manner with Fufangkushen's concentrations ranging from 1/5 to 1/400. γδT cells cytotoxic activity to SGC-7901 induced by Fufangkushen for 24 h was higher than control group, (83.6% vs 71.2%, P<0.05). Apoptotic rate was significantly lower in γδT cells than in SGC-7901 (4.64% vs49.23%, P<0.05). Conclusion Fufangkushen, within routine concentration ranges, can promote γδT cells' proliferation, inhibit tumor cell growth and enhance γδT cells' cytotoxic activity. This may be beneficial to tumor adoptive immunotherapy and provide evidence for the appli-cation of Fufangkushen in the treatment of tumors.     

维生素E琥珀酸酯诱导人胃癌细胞分化的作用

目的研究维生素E琥珀酸酯(VES)对胃癌细胞的诱导分化作用。方法5，10μg／ml维生素E琥珀酸酯处理人胃癌SGE-7901细胞，分别采用噻唑蓝(MTT)法检测细胞的生长；^3H-TdR掺入法测定DNA合成速率；流式细胞术检测细胞周期的分布；分光光度法检测分化标志酶的活性。结果VES可使细胞形态学改变；抑制SGC-7901细胞的生长。降低DNA的合成速率；使细胞出现G0／G1期停滞；分化标志酶碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)活性降低。结论VES可诱导SGC-7901细胞分化并抑制其恶性生长。     

巴豆生物碱对人胃癌细胞SGC-7901的诱导分化作用研究

以人胃癌细胞SGC-7901为研究材料,胃细胞分化标志酶碱性磷酸酶(ALP)和乳酸脱氢酶(LDH)为实验指标,观察人胃癌细胞SGC-7901在含有不同剂量巴豆生物碱培养液中的生长情况,结果发现经不同剂量巴豆生物碱处理的细胞,其碱性磷酸酶(ALP)和乳酸脱氢酶(LDH)的活力显著低于培养相同天数而未用巴豆生物碱处理的对照胃癌细胞,显示了巴豆生物碱对胃癌细胞具有诱导分化作用.     

缺氧条件下大豆异黄酮对胃癌细胞增殖和缺氧诱导因子-1α表达的影响

目的:研究缺氧条件下大豆异黄酮(soy bean isoflavone,SI)对人胃癌细胞株SGC-7901生长抑制作用及可能机制。方法:运用产气袋法(GasPaK)制造缺氧条件,实验分为正常对照、SI、缺氧对照和缺氧SI四组,用MTT法检测常氧和缺氧条件下药物效应。流式细胞仪分析各组细胞周期分布,透射电镜观察细胞超微形态的改变,免疫细胞化学法检测各组细胞缺氧诱导因子HIF-1α和FAS的表达。结果:不同浓度SI在常氧和缺氧条件下都对人胃癌细胞株SGC-7901生长有抑制作用,并呈剂量-效应关系,细胞均滞留于G2/M期,缺氧条件下SI组抑制作用增加,细胞凋亡率增加。透射电镜观察常氧下SI处理后细胞出现凋亡,缺氧条件下出现凋亡和坏死。SI在不同氧条件下均上调Fas表达,并能下调因缺氧表达增加的HIF-1。结论:SI对人胃癌细胞生长有抑制作用,其机制可能通过阻抑细胞周期进程,上调FAS蛋白表达,从而诱导胃癌细胞凋亡而实现的。缺氧条件下SI抑制作用增强,抑制HIF-1α的表达可能是其重要机制。     

灰树花多糖硫酸酯的制备及其抗肿瘤活性

利用碱提酸沉淀法从灰树花(Grifola frondosa)发酵菌丝体中获得的碱提水不溶性多糖GAP,经氯磺酸-吡啶法硫酸化修饰后,即制得能溶于水的硫酸化灰树花多糖.通过DEAE-Sepharose F.F.离子交换柱色谱分离得到一种相对分子量为2.8×104、硫酸基含量为16.4%的硫酸酯化灰树花多糖(S-GAP).S-GAP红外光谱分析表明,在1230 cm-1、810 cm-1有硫酸酯键的特征吸收峰,说明多糖已形成硫酸酯.在体外S-GAP对S-180肿瘤细胞无抑制作用,但对人胃癌细胞SGC-7901有直接杀伤作用.     

鸦胆子油乳抗人胃腺癌增殖作用的初步研究

目的 研究鸦胆子油乳对人胃腺癌细胞的增殖抑制作用。方法 采用体外药物敏感试验和流式细胞仪技术检测鸦胆子油乳对SGC－7901细胞增殖周期影响及诱导凋亡作用。结果 鸦胆子油乳体外具有明显的抑制SGC－7901增殖的作用。与对照组相比,鸦胆子油乳2μg/ml分别孵育SGC-7901细胞24h和36h后,细胞周期被阻滞于DNA合成期（S期）。流式细胞仪检测未见明显的亚二倍体峰,鸦胆子油 同浓度、不同孵育时间诱导细胞凋亡的作用无显著差异（P＞0.05）。结论 鸦胆子油乳在体外能有效抑制SGC－7901细胞的增殖,初步研究表明其主要通过阻滞细胞周期于S期而不是通过诱导凋亡来抑制SGC－7901细胞的增殖。     

Vibrio vulnificus cytolysin induces apoptosis in HUVEC,SGC-7901 and SMMC-7721 cells via caspase-9/3-dependent pathway

Vibrio vulnificus cytolysin (VVC) is known to be a pore-forming toxin which shows cytotoxicity for mammalian cells in culture and induces apoptosis in endothelial cells. In order to determine whether VVC induces apoptosis in vascular endothelial cells and tumor cells, the cytotoxicity induced by recombinant VVC (rVVC) and its potential mechanism in HUVEC, SGC-7901 and SMMC-7721 cells were investigated. Our study demonstrated that rVVC induced the release of intracellular K⁺ from all the target cells, yet lactate dehydrogenase was not released by rVVC. It indicates that osmotic lysis might not contribute to the cytolysin-induced cytotoxicity. The study also demonstrated that rVVC induced apoptosis in HUVEC, SGC-7901 and SMMC-7721 cells in time- and dosage-dependent manners, which was associated with the activation of caspase-9 and -3, but not caspase-8. During the apoptotic process of the target cells, rVVC labeled with FITC was monitored to attach initially to the surface of the cells and entered the cytoplasma subsequently. These findings suggest that VVC may be not only a pore-forming toxin, but also a transmembrane toxin with powerful ability to induce apoptosis in human vascular endothelial cells and tumor cells.     

HMGA2在胃癌细胞SGC-7901上皮间质转化中的作用

目的 探讨TGF-β1调控人胃癌细胞SGC-7901由上皮向间质细胞转化过程中,HMGA2的表达及其作用。方法 用TGF-β1处理人胃癌细胞SGC-7901(0,3,6,24,48,72h),在显微镜下观察SGC-7901细胞形态的变化,蛋白质印记法检测HMGA2、E-cadherin和Vimentin蛋白的表达,Transwell小室实验检测细胞侵袭能力。结果 (1)随着TGF-β1作用时间的延长,SGC-7901细胞逐渐变长,变大,细胞融合;(2)在TGF-β1作用下,SGC-7901均不同程度地表达HMGA2、E-cadherin和Vimentin。HMGA2蛋白从0h开始表达,到6h表达最强,24h以后表达稍有下降。E-cad蛋白从0h开始表达,到3h表达最高,随后开始降低。Vimentin蛋白48h表达最高,72h有降低的趋势;(3)Transwell小室实验显示SGC-7901在TGF-β1处理后,3h时细胞穿膜最多,从6h起穿过的细胞开始降低,到72h最低。结论 在TGF-β1诱导人胃癌细胞SGC-7901细胞上皮间质转化的过程中,HMGA2蛋白起到重要作用,HMGA2可作为Snail的启动子,结合Smads,促进上皮间质转化HMGA2还可能通过调控Wnt/β-Catenin信号通路对肿瘤细胞的生长和凋亡发挥作用。     

Inhibitory effects of dobutamine on human gastric adenocarcinoma

AIM:To explore the inhibitory effects of dobutamine on gastric adenocarcinoma cells.METHODS:Dobutamine was used to treat gastric adenocarcinoma cells(SGC-7901)and cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.The effects of dobutamine combined with cisplatin on cell viability were also analyzed.Cell migration was studied using the wound healing assay,and cell proliferation was analyzed using the colony formation assay.A cell invasion assay was carried out using Transwell cell culture chambers.The cell cycle and cell apoptosis were analyzed by flow cytometry.Western blot and immunocytochemistry were performed to determine the expression of Yes-associated protein(YAP)in treated cells.RESULTS:Dobutamine significantly inhibited cell growth,migration,cell colony formation,and cell invasion into Matrigel.Dobutamine also arrested the cell cycle at G1/S phase,and increased the rate of apoptosis of gastric adenocarcinoma cells.The expression ofYAP was detected mainly in the nucleus in the absence of dobutamine.However,reduced expression of phosphorylated YAP was mainly found in the cytosol following treatment with dobutamine.CONCLUSION:Dobutamine has significant inhibitory effects on gastric adenocarcinoma cells and may be used in neoadjuvant therapy not only for gastric cancer,but also for other tumors.