僧帽牡蛎天然活性多肽BPO-1抗人胃腺癌BGC-823细胞活性研究

运用细胞生物学、生物化学等手段 ,观察鉴定分离提取的牡蛎天然活性肽BPO 1对人胃腺癌细胞的生物学效应 .实验结果表明 ,BPO 1能有效抑制人胃腺癌BGC 82 3细胞增殖活动 ,细胞生长受到抑制 ,分裂指数下降 .细胞周期检测出现凋亡峰 .光镜观察显示经BPO 1处理后的BGC 82 3细胞失去原有的恶性表型 ,表现出明显不同于对照组细胞 .

新型TopoⅠ抑制剂CPUY013对胃腺癌细胞BGC823的体内外作用

探讨CPUY013在体内外对人胃腺癌BGC823细胞的抗肿瘤活性及其机制。采用MTT法、克隆原形成法检测CPUY013对BGC823细胞增殖的抑制作用;体外实验以pBR322DNA为底物,依据质粒的松弛和超螺旋形式在琼脂糖电泳上泳动度对药物抑制解旋的作用进行定性分析。用AO/EB荧光染色技术、DNA凝胶电泳及JC-1线粒体膜电位检测试剂盒检测细胞凋亡。口服给药CPUY013后观察其对裸鼠移植瘤的生长抑制作用;用流式细胞术观察CPUY013对BGC823细胞周期的影响;采用Western blotting分别检测CPUY013处理后的BGC823细胞中TopoⅠ及凋亡相关蛋白表达的改变。结果表明,CPUY013呈明显的剂量依赖性抑制BGC823细胞的增殖。随着剂量的增加,CPUY013对TopoI松弛活性的抑制趋势增强。100μmol·L-1CPUY013和拓扑替康(TPT)对TopoⅠ松弛活性的抑制呈现相似的变化趋势。CPUY013可使大部分BGC823细胞阻滞在S期,并诱导细胞凋亡;出现DNA片段化,亚G1峰显著增加,线粒体膜电位降低,荧光染色后出现凋亡细胞的特征性改变等。同时,CPUY013能明显抑制BGC823裸鼠移植瘤的生长,150mg·kg-1剂量组的瘤重抑制率为62.1%。CPUY013使BGC823细胞内TopoI和bcl-2蛋白表达均有所下调,p53和bax蛋白表达有明显上调趋势,bcl-2/bax比值明显降低,caspase-3蛋白表达增高。新型TopoⅠ抑制剂CPUY013在体内明显抑制移植瘤的生长,体外可诱导细胞凋亡从而抑制BGC823细胞增殖,其机制可能与抑制TopoI,从而下调bcl-2,上调bax和p53蛋白的表达有关。

大蒜素对人胃腺癌BGC-823细胞影响的研究

目的 :探讨大蒜素诱导胃癌BGC - 82 3细胞凋亡的作用及机制。 方法 :传代培养人胃腺癌BGC - 82 3细胞 ,使细胞浓度为 5× 10 5个 /ml。分别加入 10 -5g/L(大蒜素 1)和 10 -3 g/L(大蒜素 2 )的大蒜素 ,另设对照组 ,培养时间分别为 6h、2 4h、48h、72h。 结果 :与对照组相比 :①大蒜素可明显提高胃癌细胞凋亡率、降低PI值 (P <0 0 1或P <0 0 5 ) ,且表现出明显的细胞凋亡的形态学变化。②大蒜素可明显降低胃癌细胞CEA、GST -π的水平 (P <0 0 1或P <0 0 5 )。③大蒜素可明显提高胃癌细胞的cAMP水平 (P <0 0 1) ,降低PKC水平 (P <0 0 1或P <0 0 5 )。④大蒜素能够显著增加胃癌细胞fas基因及bax基因蛋白的表达 (P <0 0 5或P <0 0 1) ;降低bcl- 2基因蛋白的表达 (P <0 0 1)。 结论 :大蒜素可通过对第二信号系统cAMP、PKC的影响 ,增强细胞凋亡始动基因fas、凋亡促进基因bax的表达 ,抑制凋亡抑制基因bcl- 2的表达 ,增加细胞凋亡率 ,并使胃癌细胞调亡 ,并抑制其增殖活性 ;并可导致胃癌细胞CEA、GST -π的合成减少 ,从而降低其水平。

蜘蛛毒素对人胃腺癌细胞株BGC-823增殖的抑制作用

为比较不同蜘蛛毒素对肿瘤细胞的抑制作用,采用四甲基偶氮唑盐比色法(MTT法)测定了不同浓度的虎纹单柄蛛(Ornithocton huwen)、海南单柄蛛(Ornithoctonus hainana)、敬钊缨毛蛛(Chilobrachy jingzhao)和雷氏大疣蛛(Macrothele raveni)4种蜘蛛毒素对人胃腺癌细胞株BGC-823的抑制作用。结果表明:4种蜘蛛毒素对人胃腺癌细胞株BGC-823的细胞增殖均有不同的抑制作用;同一蜘蛛毒素浓度越高抑瘤作用越好,蜘蛛毒素作用时间越长抑瘤效果越强,量效关系和时效关系良好;不同蜘蛛毒素对BGC-823细胞增殖的抑制作用效果不同,以敬钊缨毛蛛毒素作用最强,IC50为0.109μg/μL。

姜黄素对人胃腺癌细胞株BGC-823生长抑制及诱导凋亡的作用

目的探讨姜黄素对人胃腺癌细胞BGC-823的生长抑制及诱导凋亡作用。方法 MTT法检测不同浓度姜黄素对人胃腺癌BGC-823细胞的生长抑制作用,计算半数抑制浓度(IC50);将1.2μmol/L姜黄素作用于BGC-823细胞24、48、72h,用DAPI染色检测细胞凋亡;用RT-PCR、Western blotting法分别检测姜黄素对Bax、Bcl-2基因以及Bax、Bcl-2、caspase-3蛋白的影响。结果不同浓度的姜黄素作用于BGC-823细胞0～72h后,细胞呈不同程度的抑制作用,随着药物浓度的增加,抑制率明显上升,作用48h的IC50为1.2μmol/L。DAPI染色可见典型的凋亡细胞形态学特征。RT-PCR、Western blotting结果显示1.2μmol/L姜黄素可以增加Bax表达、减少Bcl-2表达和激活caspase-3。结论姜黄素对人胃腺癌BGC-823细胞有生长抑制和诱导凋亡的作用,这可能与增加Bax表达、减少Bcl-2表达、激活caspase-3有关。

人胃腺癌BGC823细胞Midkine高表达前后基因表达谱芯片分析

目的:用基因芯片技术分析、筛选Midkine高表达前后胃腺癌相关基因谱的差异表达。方法:建立Midkine高表达BGC823胃腺癌细胞系,提取细胞总RNA,逆转录Cy3、Cy5标记cDNA探针,应用晶芯表达谱芯片,分析差异表达的基因。结果:发现Midkine高表达前后BGC823胃腺癌细胞有显著性表达差异的基因共有550个,其中上调基因407个,下调基因143个。对550条差异表达基因进行初步功能分类分析,结果表明这些基因与Midkine促进胃腺癌发病机制存在相关性。结论:Midkine高表达前后BGC823胃腺癌细胞部分功能基因存在显著的表达差异,多数差异表达的基因参与细胞黏附、细胞骨架形成、细胞周期以及细胞代谢等过程。

rh-bFGF抑制顺铂诱导的人胃腺癌BGC-823细胞凋亡

目的 :评价外源加入的rh -bFGF对抗肿瘤药物顺铂所致人胃腺癌细胞凋亡的影响 方法 :利用顺铂诱导人胃腺癌BGC - 82 3细胞产生的凋亡作模型 ,体外培养时在顺铂作用前或作用后加入不同浓度的rh -bFGF ,观察细胞形态的变化和细胞凋亡比率的变化 结果 :预培养时rh -bFGF质量浓度为 10ng/mL及 1ng/mL时抑制细胞凋亡的发生 ;共培养时则在质量浓度为 1ng/mL时产生抑制凋亡的作用 结论 :外源加入的rh -bFGF在一定的浓度范围内 ,不论预培养还是与顺铂的共培养 12h ,均可不同程度的抑制顺铂诱导的人胃腺癌BGC - 82 3细胞的凋亡 ,并且这种剂量效应与rh -bFGF影响BGC - 82

姜黄素对人胃癌BGC823细胞的生物学作用

20世纪80年代以来已陆续发现姜黄素具有抗氧化、抗炎、抗凝和降血脂作用，并发现姜黄素还具有显著的防癌和抑癌作用。因此，它已受到人们日益关注并被广泛研究。

Induction of apoptosis and G2/M cell cycle arrest by oridonin in human gastric cancer BGC-823 cells

Aim To investigate in vitro apoptosis-induction effects of oridonin on gastric tumor cells BGC-823 and its effects on cell cycle, mitochondrial membrane potential and intracellular Ca^2＋ to shed light on the mode of its anticancer action. Methods The MTT method was used to investigate the inhibitory effect of oridonin on BGC-823 cells. The apoptosis-induction effect was evaluated by confocal laser microscopy and flow cytometry. The change of mitochondrial membrane potential and the increase of intracellular Ca^2＋ were assessed by fluorescence probe rhodamine123 and Fluo 3-AM, respectively, with flow cytometry. The expression of apoptosis and cell cycle related proteins was studied using western blotting. Results Oridonin inhibited BGC-823 cells growth with IC50 of 22.21 p, mol.L^-1. It induced apoptosis in a dose-dependent manner. In addition, it decreased mitochondria membrane potential, increased intracellular Ca^2＋, and activated pro-caspase 3. BGC-823 cells were arrested in G2/M cell cycle phase with lower expression of cyclin A protein. The up-regulation of p53 was observed before apoptosis and cell cycle arrest occurred. Conclusion Oridonin inhibits the proliferation of BGC-823 cells through G2/M cell cycle arrest and apoptosis induction, which is mediated by influx of Ca^2＋, up-regulation of p53, activation of caspase-3, and down-regulation of cyclin A.

Anticancer effect and apoptosis induction of gambogic acid in human gastric cancer line BGC-823

AIM: To investigate the anticancer effect of a traditional Chinese medicine gambogic acid (GA) in human gastric cancer line BGC-823 and further study the mechanism of apoptosis induction of GA.METHODS: Low differential human gastric cancer line BGC-823 were treated with GA at different doses and different times, the inhibitory rates were detected by MTT assay. Apoptosis induced by GA in BGC-823 cells was observed by Annexin-V/PI doubling staining flow cytometry assay. And T/C (%) was chosen to detect the inhibition of GA on human gastric adenocarcinoma BGC-823 nude mice xenografts. Apoptosis on nude mice xenografts was observed by Annexin-V/PI doubling staining flow cytometry assay and DNA fragmentation assay. To further determine the molecular mechanism of apoptosis induced by GA, the changes on the expression of bcl-2 and bax genes were detected by RT-PCR.RESULTS: After incubation with GA, low differential human gastric cancer line BGC-823 was dramatically inhibited in a dose-dependent manner. After these cells were exposedto GA for 24, 48 and 72 h, the IC50 value was 1.02±0.05, 1.41±0.20 and 1.14±0.19 μmol/L, respectively. Apoptosis in BGC-823 cells induced by GA was observed by AnnexinV/PI doubling staining flow cytometry assay. The apoptotic population of BGC-823 cells was about 12.96% and 24.58%, respectively, when cells were incubated with 1.2 μmol/L GA for 48 and 72 h. T/C (%) of human gastric carcinoma adenocarcinoma BGC-823 nude mice xenografts was 44.3, when the nude mice were treated with GA (8 mg/kg). Meanwhile, apoptosis induced by GA was observed in human gastric carcinoma adenocarcinoma BGC-823 nude mice xenografts. The increase of bax gene and the decrease of bc1-2 gene expressions were found by RT-PCR.CONCLUSION: The inhibition of GA on human gastric cancer line BGC-823 was confirmed. This effect connects with the inducing apoptosis in BGC-823 cells and the molecular mechanism might be related to the reduction of expression of apoptosis-regulated gene bcl-2, and the improvement of the expression of apoptosis-regulated gene bax. The result was also confirmed in vivo.

Co-expression of heat shock protein 70 and glucose-regulated protein 94 in human gastric carcinoma cell line BGC-823

AIM:To investigate the co-expression and significance of heat shock protein 70 (HSP70) and glucose-regulated protein 94 (grp94) in human gastric carcinoma cell line BGC-823. METHODS: The expression and localization of HSP70 and grp94 in human gastric carcinoma cell line BGC-823 were determined by immunocytochemistry and indirect immunofluorescence cytochemical staining. Flow cytometry was used to analyze the correlation between expression of HSP70, grp94 and cell cycle in BGC-823 cell line. RESULTS: Gastric cancer cell line BGC-823 expressed high level of HSP70 and grp94. The positive rate of HSP70 and grp94 was 84.9±4.94% and 79.6±5.16%, respectively. Both of them were stained in cell plasma. There was a significant difference compared with control group (1.9±0.94%, P<0.01). During the cell cycle, HSP70 and grp94 were continuously expressed in BGC-823. CONCLUSION: HSP70 and grp94 are highly expressed in human gastric carcinoma BGC-823 cells through the whole cell cycle. There is no relationship between expression of HSP70, grp94 and cell cycle.

Effect of IL-12 on the proliferation and cytotoxicity of CIK cells to gastric adenocarcinoma cell BGC-823

Objective: The aim of the study was to evaluate the effect of interleukin-12(IL-12) on the proliferation and cytotoxicity of cytokine-induced killer(CIK) cells in vitro. Methods: Three different combinations of cytokines, IL-2, IL-12 + IL-2, and IL-12, were used to proliferate CIK cells, adding IFN-γ, IL-1 and CD3 McAb in each one. Phenotype of the cells was analyzed by flow cytometry. The cellular proliferation and cytotoxic activity were determined by cytometry and MTT assay. Results: CIK cells generated by the three methods showed high reproductive activity, and no obviously difference in inducing CD3+CD56+ cells was found among the three groups. The group of IL-2 + IL-12 evidently enhanced both the proliferation and the cytotoxicity of the CIK cells compared with the other two groups(P < 0.05). Conclusion: IL-12 could be used to induce the CIK cells as well as IL-2. CIK cells induced by combining IL-12 with IL-2 had stronger proliferative ability and higher cytotoxicity to tumor cells in vitro, which could be used as a potential anti-tumor adoptive immunotherapy in clinic.

穿心莲内酯体外对人胃腺癌BGC823细胞增殖凋亡的影响

目的探讨穿心莲内酯（andrographolide，AD）对人胃癌细胞株BGC-823细胞增殖、细胞周期以及细胞凋亡的影响。方法分别采用MTT法、流式细胞术和流式细胞仪AnnexinV／PI双染色法检测AD对BGC-823细胞体外增殖、细胞周期和细胞凋亡的影响：应用光镜和透射电镜观察不同浓度的AD作用后BGC．823细胞形态学改变。结果各浓度组AD均对人低分化胃癌细胞株BGC-823的增殖有抑制作用。并具有时间和浓度依赖关系（均P〈0．05）。浓度7．5μg／ml以下的AD抑制效果较弱，而15．0-60．0μg／ml抑制效果显著提高（P〈0．05），60．0μg／ml以上抑制率增高不显著（P〉0．05）。24、48和72h的IC50分别为（35．3±4．3）、（25．5±3．5）和（18．2±2．7）μg／ml。BGC-823细胞经AD作用后，G0／G1期细胞的比例增加，S期和G2/M期细胞的比例下降，细胞被阻滞在G0／G1期，呈浓度依赖关系。AD浓度为7．5、10．0和15．0μg／ml组作用24h后，早期凋亡率分别为（19．3±4．7）％、（29．4±4．1）％和（52．7±6．7）％，晚期凋亡率为（10．8±1．8）％、（10．9±4．7）％和（14．7±4．8）％，均显著高于阴性对照组的早期凋亡率[（3．4±1．0）％]和晚期凋亡率[（4．1±0．7）％]，差异有统计学意义（均P〈0．05），并呈浓度依赖关系。结论AD能抑制BGC-823细胞增殖、阻滞其细胞周期在G0／G1期和诱导其细胞凋亡．是潜在的胃癌抗肿瘤中药制剂成分。

黄药子体外抗胃癌活性成分的筛选及分析

目的通过MTT体外实验观察黄药子4种不同提取物及从石油醚提取物中分离得到的黄独乙素和β-扶桑甾醇作用于MGC803、BGC823和SGC7901 3种人胃癌细胞株,探讨黄药子不同提取物及两个单体对3种胃癌细胞株的增殖情况的影响,从而筛选出黄药子有效抗胃癌活性成分。方法采用MTT法检测0.5、1、2、4、6、8、10、12 mg/mL的不同质量浓度的黄药子石油醚萃取物、乙酸乙酯萃取物、水层萃取物、醇提物和最终质量浓度为1×10-4、5×10-4、1×10-3、5×10-3、1×10-2、5×10-2、1×10-1、5×10-1mg/mL的黄独乙素和β-扶桑甾醇分别作用于MGC803、BGC823和SGC 7901细胞株24 h后对细胞增殖的影响。结果石油醚萃取物、醇提物、黄独乙素和β-扶桑甾醇均有不同程度的抑制肿瘤细胞生长的作用,而乙酸乙酯萃取物和水萃取物低剂量有不同程度的促进肿瘤细胞生长的作用;大剂量表现为抑制肿瘤细胞生长的作用。结论从石油醚提取物中分离得到的黄独乙素和β-扶桑甾醇对胃癌细胞均有抑制作用。

伊立替康联合去甲斑蝥素对人胃癌细胞作用的考察及协同作用机制研究

研究了伊立替康(CPT-11)联合去甲斑蝥素(NCTD)对人胃癌细胞BGC-823增殖的影响及协同作用机制.分别利用CPT-11 30、60、90、120、150μmol/L,NCTD 30、60、90、120、150μmol/L和两药按上述浓度1∶1联用,以及两药上述浓度完全交叉组合作用BGC-823细胞24、48和72 h,并且以不同序贯方式作用BGC-823细胞24 h.MTT法检测BGC-823细胞的增殖,采用中效原理评价两药联合作用;流式细胞术测定CPT-11 60μmol/L、NCTD 60μmol/L和联合用药(60∶60)μmol/L,以及不同序贯方式作用BGC-823细胞24 h后细胞周期及凋亡的情况;Western blot法检测CPT-11 30、60μmol/L,NCTD 30,60μmol/L和联合用药(30∶30,60∶60)μmol/L作用BGC-823细胞24 h后Pdcd4和p53蛋白表达水平.结果显示:CPT-11及NCTD联合用药比单独用药对细胞增殖的抑制作用增强,IC50明显减小(P<0.05),单独使用CPT-11或NCTD作用24、48和72 h时的IC50分别是联合使用时的2.83、3.15、2.19倍以及2.66、3.11、2.45倍,并且两药联合用药具有协同作用效应;两药合用24 h时先给CPT-11方案抑制作用优于先给NCTD方案,且优于同时给药方案(P<0.05);CPT-11 60μmol/L作用24 h引起BGC-823细胞S和G2-M期阻滞(P<0.01),NCTD 60μmol/L作用24 h引起细胞G2-M期阻滞(P<0.05),并诱导细胞凋亡(P<0.05),联合用药(60∶60)μmol/L作用24 h,主要引起细胞G2-M期阻滞(P<0.01),与先给NCTD 6h方案及同时给药相比,先给CPT-11 6h方案主要引起细胞S期阻滞增加(P<0.05)以及细胞凋亡增加;CPT-11 30、60μmol/L作用12 h后Pdcd4上调表达,p53下调表达(P<0.05),NCTD 30、60μmol/L作用12 h后Pdcd4下调表达,p53上调表达(P<0.05),联合用药(30∶30,60∶60)μmol/L作用24 h后Pdcd4及p53蛋白表达均上调(P<0.05).上述结果表明:CPT-11联合NCTD对人胃癌BGC-823细胞有协同抑制作用,其机制主要是诱导细胞G2-M期阻滞,并与抑癌基因蛋白Pdcd4及p53上调表达有关;两种药物联用时序贯次序对联合作用有影响,先给CPT-11方案要优于先给NCTD方案及同时给药方案,其机制主要是引起S期阻滞增加以及细胞凋亡增加;抑癌基因蛋白Pdcd4与p53之间可能存在负调控关系.

RASSF6介导凋亡与自噬提高奥沙利铂对胃癌细胞的化疗敏感性

目的探讨RASSF6基因对胃癌细胞增殖、自噬、凋亡及对奥沙利铂化疗敏感性的影响。方法体外培养胃癌BGC823细胞,依据慢病毒基因表达分为正常对照组(control组)、RASSF6过表达组(Oe组)、RASSF6干扰组及慢病毒对照组。分别检测各组细胞增殖、细胞周期分布、细胞迁移、自噬、凋亡变化及对奥沙利铂化疗的敏感性,电镜观察各组细胞自噬小体数,荧光定量PCR(qRT-PCR)和蛋白免疫印迹(Western blotting)检测各组细胞凋亡和自噬相关基因的表达。结果 RASSF6基因表达对胃癌BGC823细胞生物学行为的影响,与control组相比,Oe组细胞G0/G1期细胞百分比升高,G2期、S期细胞百分比降低,差异有统计学意义(P<0.05);细胞凋亡率明显增加,差异有统计学意义(P<0.05);细胞划痕面积划痕愈合率显著下降,差异均具有统计学意义(P<0.05)。RASSF6基因表达对奥沙利铂药物敏感性的影响,与药物组(L-OHP组)相比,Oe+L-OHP组细胞凋亡率升高,差异有统计学意义(P<0.05);Western blotting检测示Oe+L-OHP组细胞抗凋亡蛋白Bcl-2表达下降,促凋亡蛋白Bax、Caspase-3表达升高,细胞自噬小体增多,自噬相关蛋白Beclin-1、P62表达及LC3-Ⅱ/LC3-Ⅰ比值升高(P<0.05)。结论 RASSF6过表达可抑制胃癌细胞增殖,促进细胞自噬和凋亡并提高胃癌细胞对奥沙利铂化疗敏感性。RASSF6基因可作为胃癌治疗新的靶点。

冬凌草甲素注射液对人胃腺癌裸鼠移植瘤的抑制作用及其机制

目的观察冬凌草甲素注射液对BGC823人胃腺癌裸鼠移植瘤的生长抑制作用，并探讨其作用机制。方法建立裸鼠BGC823人胃腺癌移植瘤模型，并随机分为空白对照组、低剂量组和高剂量组，分别通过尾静脉注射生理盐水、冬凌草甲素注射液37．5mg·ks^-1·d^-1和冬凌草甲素注射液75nag·ks^-1·d^-1。10d后，剥出瘤块称移植瘤重量，光镜及电镜下观察移植瘤组织形态学及瘤细胞的超微结构变化，免疫组化SABC法检测移植瘤bcl-2、Bax、Fas和FasL蛋白的表达差异。结果冬凌草甲素注射液可抑制裸鼠BGC823人胃腺癌移植瘤的生长，低剂量组与高剂量组的抑瘤率分别为48．5％和70．7％。病理形态学显示，冬凌草甲素注射液作用后移植瘤出现细胞凋亡及组织坏死。免疫组化检测显示，冬凌草甲素注射液作用后瘤组织中bcl-2蛋白表达减少，Bax、Fas和FasL蛋白表达增加。结论冬凌草甲素注射液可显著抑制BGC823人胃腺癌裸鼠移植瘤的生长，其作用机制与诱导肿瘤细胞凋亡有关。

地花菌提取物Albaconol的抗肿瘤活性研究

目的:研究地花菌提取物albaconol的抗肿瘤活性。方法:以人白血病K 5 62、人乳腺癌Bcap 3 7、人胃腺癌BGC 82 3和人非小细胞肺癌A5 49为模型,采用MTT法测试albaconol对肿瘤细胞增殖的影响;以小鼠肉瘤S180和小鼠肝癌H2 2为移植性肿瘤模型,检测albaconol静脉给药对肿瘤生长的影响。结果:Albaconol对K5 62、Bcap 3 7、BGC 82 3、A5 49的IC50 分别为2 .42±1.77、1.88±1.41、1.0 4±0 .64、1.18±1.10 μg/ml ;Albaconol 0 .87、1.73、3 .46mg/kg剂量组对S180的抑瘤率分别为2 8.2 %、43 .2 %、47.4% ,对H2 2的抑瘤率分别为15 .6%、2 2 .4%、3 7.8%。结论:Albaconol具有较强的体内、外抗肿瘤活性。

小半夏加茯苓醇提物对HepG2、BGC823线粒体凋亡途径的影响

目的探讨小半夏加茯苓醇提物对人肝癌细胞HepG2和人胃腺癌细胞BGC823增殖抑制作用及线粒体凋亡途径的影响。方法运用CCK-8法检测HepG2、BGC823的增殖抑制情况,计算生长抑制率及半数抑制浓度;运用流式细胞术和实时荧光定量PCR检测HepG2、BGC823的线粒体跨膜电位和bax、cjun基因的表达情况。结果小半夏加茯苓醇提物对HepG2、BGC823均有显著的抑制作用(P<0.01),其半数抑制浓度分别为(781.50±13.00)μg/mL和(560.05±10.06)μg/mL;与空白对照组比,试验组HepG2的荧光强度升高(P<0.05),试验组BGC823的荧光强度显著升高(P<0.01);HepG2、BGC823试验组的bax、cjun基因的相对表达量与空白对照组比较,均显著升高(P<0.01)。结论小半夏加茯苓醇提物诱导HepG2、BGC823凋亡的机制可能与激活线粒体途径有关。

Gastric cancer cell lines induced by trichostatin A

AIM： To explore the effect of trichostatin A （TSA） on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS： The effect of TSA on growth inhibition and apoptosis was examined by MTT, fluorescence microscopy and PI single-labeled flow cytometry. The acetylated histone H3 level was detected by Western blot. RESULTS： TSA induced apoptosis in gastric cancer cell lines BGC-823 and SGC-7901 was in a dose and time-dependent manner. Apoptotic cells varied significantly between TSA treated groups （37.5 ng/mL 72 h for BGC-823 cell line and 75 ng/mL 72 h for SGC-7901 cell line） and control group （0.85 ± 0.14 vs 1.14 ± 0.07, P = 0.02; 0.94 ± 0.07 vs 1.15 ± 0.06, P = 0.02）. Morphologic changes of apoptosis, including nuclear chromatin condensation and fluorescence strength, were observed under fluorescence microscopy. TSA treatment in BGC-823 and SGC-7901 cell lines obviously induced cell apoptosis, which was demonstrated by the increased percentage of sub-G1 phase cells, the reduction of Gl-phase cells and the increase of apoptosis rates in flow cytometric analysis. The result of Western blot showed that the expression of acetylated histone H3 increased in BGC-823 and SGC-7901 TSA treatment groups as compared with the control group.CONCLUSION： TSA can induce cell apoptosis in BGC-823 and SGC-7901 cell lines. The expression of acetylated histone H3 might be correlated with apoptosis.