Licochalcone A Induces Cholangiocarcinoma Cell Death Via Suppression of Nrf2 and NF-κB Signaling Pathways

Objective: To investigate the anti-tumor effect of licochalcone A (LCA) on proliferation and migration in cholangiocarcinoma (CCA) cells and to elucidate their underlying mechanisms. Methods: Human CCA cells, KKU-100, KKU-213, KKU-214, KKU-156, and KKU-452 were used to study effect of LCA on proliferation and migration by a cytotoxicity assay, wound healing assay. Reactive oxygen species levels were evaluated using DHE-fluorescent probes. Proteins associated with cancer survival and progression were analyzed by immune blotting assay. Results: LCA suppressed proliferation and induced cell death in CCA cells including KKU-100, KKU-213, KKU-214, KKU-156, and KKU-452. The CCAs cells were suppressed in association with LCA-induced accumulation of intracellular reactive oxygen species (ROS). Increased formation of ROS was causally related with suppression of Nrf2 and its down-stream antioxidant and cytoprotective enzymes. These effects may lead to the expression of Bax and release of cytochrome c and ensuring cell death.  Interestingly, LCA could also inhibit cell migration and cell cycle arrest at low concentrations. These effects were associated with down-regulation of NF-kB, STAT3 and their down-stream proteins, cyclin D1, VEGF, and ICAM-1. Conclusions: These results suggest that LCA has potential therapeutic activity in suppression of CCA cells.     

阿司匹林抑制肺癌细胞增殖的实验研究

 目的 观察阿司匹林在体外对肺腺癌细胞A549增殖的抑制作用。方法 采用噻唑蓝（MTT）法观察阿司匹林及与奥沙利铂联用抑制A549细胞的增殖；采用流式细胞仪（FCM）观察阿司匹林处理后A549细胞周期分布的变化；采用HE染色和DNA末端原位标记染色技术（TUNEL）观察阿司匹林处理后诱导A549细胞凋亡的作用。结果 MTT显示阿司匹林呈剂量依赖方式抑制A549细胞增殖。阿司匹林作用后，FCM显示G0／G1期细胞比例增加，S期和G／M期细胞比例降低，TUNEL显示细胞凋亡指数（AI）由3．67％±1．15％增加到26．33％±2．52％，呈一定剂量效应关系。光镜下可见典型的细胞凋亡形态学变化。阿司匹林（2．5mmol／L）与奥沙利铂（≥6．25ug／mL）联用可增强抑制A549增殖的作用，二者呈协同或相加作用。结论 阿司匹林可抑制肺腺癌细胞A549的增殖，其影响细胞周期分布、诱导细胞凋亡可能是其重要的机制。阿司匹林及与奥沙利铂联用有显著的协同抗增殖效应。     

下调SMAD1表达诱导胃癌细胞周期进展及促进细胞增殖抑制细胞凋亡的作用

目的:探讨下调SMAD1表达对胃癌细胞周期、增殖及凋亡的影响。方法:将胃癌BGC-823细胞分为三组,以转染SMAD1 siRNA的细胞为SMAD1 siRNA组,转染SMAD1 control细胞为SMAD1 NC组,对照组不做任何处理。采用Western blot检测细胞的转染效果及PTEN、Bcl-xL 和p21蛋白的表达情况,CCK-8法检测细胞的增殖情况,流式细胞仪检测细胞周期变化和细胞凋亡率。结果:转染48 h后,SMAD1 siRNA组细胞中SMAD1蛋白的相对表达量显著降低（P＜0.05）;细胞在G0/G1期所占比例显著降低,S期所占比例显著升高;SMAD1 siRNA组细胞的OD值、Bcl-xL蛋白的相对表达量显著升高,而细胞凋亡率及PTEN、p21蛋白的相对表达量显著降低,差异均有统计学意义（P＜0.05）。结论:下调SMAD1表达能够诱导BGC-823细胞从G0/G1期进入S期,促进细胞增殖并抑制细胞凋亡,其作用机制可能与下调PTEN、p21蛋白表达及上调Bcl-xL蛋白表达有关。     

舒林酸对胃癌细胞株SGC7901细胞周期调控及凋亡的影响

目的观察舒林酸影响胃癌细胞株SGC7901细胞周期分布,诱导细胞凋亡,探讨其作用机制方法采用MTT比色法观察舒林酸对胃癌SGC7901细胞增殖的影响,采用流式细胞仪检测舒林酸对细胞周期分布的影响,同时结合透射电子显微镜观察舒林酸诱导SGC901细胞凋亡的作用;免疫细胞化学方法观察舒林酸对胃癌细胞周期调控蛋白cyvclinE及P21WAF/CIPI的影响结果舒林酸可抑制胃癌SGC7901细胞的增殖,改变细胞周期分布,使G-0/G1期细胞比例增高,S期比例降低,并对SGC7901细胞有促凋亡作用.上述作用具有剂量和时间依赖性(P＜0.05).舒林酸还可上调P21WAF/CIPI蛋白表达.下调cyolinE蛋白表达.结论舒林酸可改变细胞周期分布,影响细胞周期调控蛋白表达,并可诱导SGC7901细胞凋亡,从而抑制细胞增殖.     

舒林酸抗胃癌细胞体外生长的实验研究

目的观察舒林酸对胃癌BGC-823细胞增殖、凋亡的影响,探讨其作用机制。方法将不同浓度的舒林酸体外作用于人胃癌BGC-823细胞,采用倒置显微镜观察BGC-823细胞形态改变,四甲基偶氮唑蓝（MTT）比色法检测胃癌细胞增殖,流式细胞仪检测胃癌细胞周期分布及凋亡,透射电镜观察凋亡,免疫组化检测ki-67、bcl-2及环氧合酶-2（COX-2）蛋白的表达。结果舒林酸作用后,细胞伪足回缩,变小、变圆,排列松散或聚集成团,出现悬浮现象,瘤巨细胞减少或不见瘤巨细胞。舒林酸可抑制BGC-823细胞增殖,使G0／G1期细胞比例增高,S期细胞比例降低。1．2mmol／L舒林酸作用48h,G0／G1期细胞增至93．8％,S期细胞比例降至3．4％。舒林酸作用后,细胞凋亡率显著上升,1．2mmol／L舒林酸作用48h,细胞凋亡率升至54．9％,而ki-67、bcl-2及COX-2蛋白表达阳性率显著降低;透射电镜可观察到细胞凋亡的形态特征及凋亡小体。上述作用均呈时间和剂量依赖性。结论舒林酸可抑制胃癌BGC--823细胞体外生长,其机制涉及影响细胞周期分布、诱导细胞凋亡及抑制ki-67、bcl-2及COX-2蛋白的表达。     

小檗胺对胃癌细胞增殖、凋亡的影响及其分子机制

目的:探讨小檗胺对胃癌细胞（AGS和SGC-7901）增殖、凋亡的影响并探讨其分子机制。方法:利用MTT 实验检测胃癌细胞的增殖,利用集落形成实验检测胃癌细胞的生长, 流式细胞技术检测胃癌细胞的凋亡率以及细胞周期, Western blot检测凋亡相关蛋白的表达水平。结果:小檗胺（0~64 μg/ml）能够剂量依赖性以及时间依赖性的抑制 AGS和SGC-7901细胞的增殖 （P＜0.05）。集落形成实验结果显示小檗胺(32 μg/ml)能够抑制AGS和SGC-7901细胞的生长 （P＜0.05）。流式细胞实验显示,小檗胺(32 μg/ml)同时能够促进AGS和SGC-7901细胞的凋亡 （P＜0.05）,增殖G0/G1期细胞比例,降低S期细胞比例 （P＜0.05）。Western blot实验结果显示小檗胺(32 μg/ml)能够增加AGS和SGC-7901细胞的cleaved caspase-3,cleaved caspase-9以及Bax的蛋白水平,同时降低Bcl-2的蛋白水平 （P＜0.05）。结论:小檗胺能够抑制胃癌细胞的增殖,其机制可能与改变细胞周期以及促进细胞凋亡有关。     

PI3K/AKT信号通路阻断药联合小檗碱对前列腺癌DU145细胞生长的抑制作用

目的 探讨采用磷脂酰肌醇3-激酶(PI3K)抑制药——LY294002抑制磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/AKT)信号通路联合小檗碱对前列腺癌DU145细胞生长的抑制作用.方法 采用MTT法检测细胞增殖情况;采用流式细胞仪检测细胞凋亡率和细胞周期分布情况;采用Western blot检测细胞中PI3K、p-AKT、cyclin D1、bcl-2、BAX蛋白表达情况.结果 随着小檗碱作用浓度的增加,DU145细胞的增殖率逐渐降低;不同浓度小檗碱与LY294002联合处理对DU145细胞增殖的抑制作用均较相同浓度小檗碱单独处理增强(P﹤0.05).与空白对照组相比,小檗碱组DU145细胞的凋亡率增高(P﹤0.05),G0/G1期细胞所占比例增高(P﹤0.05),PI3K、p-AKT、cyclin D1、bcl-2蛋白表达水平均下调(P﹤0.05),BAX蛋白表达水平上调(P﹤0.05).与小檗碱组和空白对照组相比,小檗碱+LY294002组DU145细胞的凋亡率增高(P﹤0.05),G0/G1期细胞所占比例增高(P﹤0.05),PI3K、p-AKT、cyclin D1、bcl-2蛋白表达水平均下调(P﹤0.05),BAX蛋白表达水平上调(P﹤0.05).结论 小檗碱能够抑制DU145细胞增殖,并促进细胞凋亡和细胞周期阻滞,采用LY294002阻断PI3K/AKT信号通路能够明显增强小檗碱对前列腺癌DU145细胞生长的抑制作用.     

A novel mechanism for aspirin-mediated growth inhibition of human colon cancer cells.

PURPOSE: The molecular mechanisms by which aspirin and other nonsteroidal anti-inflammatory drugs exert chemopreventative effects in colon cancer are unclear and complex. Current investigations focus on the chemopreventive properties of nonsteroidal anti-inflammatory drugs, independent of their ability to inhibit cyclooxygenase (COX) activity, and presumably, identification of non-COX pathways will suggest new targets for clinical use. It was demonstrated recently that aspirin results in reduced microsatellite instability in colorectal cancer cells. We hypothesized that aspirin treatment might alter expression of DNA mismatch repair (MMR) proteins, representing another potential non-COX mechanism for its action. EXPERIMENTAL DESIGN: In this study, we have examined the effects of aspirin on the cellular growth rates, MMR protein levels, cell cycle analysis and apoptosis in MMR-deficient (HCT116) and MMR-proficient (HCT116+chr3 and SW480) human colon cancer cell lines. RESULTS: We found that treatment with aspirin inhibited the growth of these three cancer cell lines. In HCT116+chr3 cells, treatment with 1 mM of aspirin increased expression of the hMLH1 and hPMS2 proteins by 2.5-fold and 2-fold, respectively, and increased expression of the hMSH2 and hMSH6 proteins by 2-3-fold. For SW480 cells, treatment with 1 and 5 mM of aspirin increased expression of the hMLH1 and hPMS2 proteins by 2-4-fold and 3-5-fold, respectively, and increased expression of the hMSH2 and hMSH6 proteins by 3-7-fold. For all three of the cell lines, treatment with 1 and 2.5 mM of aspirin induced apoptosis at 48 and 72 h. Aspirin induced G(0)/G(1) cell cycle arrest in HCT116 cells. CONCLUSIONS: We conclude that aspirin acts through COX-independent mechanisms by resulting in an increase in MMR protein expression and subsequent apoptosis, which might serve as an additional means of growth inhibition in aspirin-treated human colon cancer cells.     

Aspirin sensitizes cancer cells to TRAIL-induced apoptosis by reducing survivin levels.

PURPOSE: Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and agonistic antibodies targeting its receptors are promising cancer therapies because of their tumor selectivity, many tumors are resistant to TRAIL-based therapies. We examined whether the nonsteroidal anti-inflammatory drug aspirin sensitized cancer cells to TRAIL agonists in vitro and in vivo and investigated the underlying mechanism. EXPERIMENTAL DESIGN: The effects of aspirin on sensitivity to TRAIL agonists and expression of apoptosis regulators was determined in human breast cancer cell lines and xenograft tumors. The specific role of survivin depletion in the TRAIL-sensitizing effects of aspirin was determined by silencing survivin. RESULTS: Aspirin sensitized human breast cancer cells, but not untransformed human mammary epithelial cells, to TRAIL-induced caspase activation and apoptosis by a cyclooxygenase-2-independent mechanism. Aspirin also sensitized breast cancer cells to apoptosis induced by a human agonistic TRAIL receptor-2 monoclonal antibody (lexatumumab). Aspirin treatment led to G1 cell cycle arrest and a robust reduction in the levels of the antiapoptotic protein survivin by inducing its proteasomal degradation, but did not affect the levels of many other apoptosis regulators. Silencing survivin with small interfering RNAs sensitized breast cancer cells to TRAIL-induced apoptosis, underscoring the functional role of survivin depletion in the TRAIL-sensitizing actions of aspirin. Moreover, aspirin acted synergistically with TRAIL to promote apoptosis and reduce tumor burden in an orthotopic breast cancer xenograft model. CONCLUSIONS: Aspirin sensitizes transformed breast epithelial cells to TRAIL-based therapies in vitro and in vivo by a novel mechanism involving survivin depletion. These findings provide the first in vivo evidence for the therapeutic utility of this combination.     

环氧化酶-2抑制剂nimesulide对乳腺癌细胞株增生、凋亡的影响

目的 探讨环氧化酶-2抑制剂nimesulide(NIM)对雌激素受体(ER)阴性(MDA-MB．231)和阳性(MCF-7)人乳腺癌细胞株增生、凋亡的影响。方法 应用MTT比色法分析细胞生长抑制作用，流式细胞技术测定细胞周期分布和凋亡率，透射电镜观察细胞形态与超微结构，AnnexinV法检测细胞的凋亡。结果 NIM以时间、剂量依赖性方式抑制MDA-MB-231(COX-2阳性)、MCF-7(COX-2阴性)细胞生长，阻滞细胞周期于G0／G1期，诱导细胞的凋亡，MDA—MB-231细胞对NIM的作用更为敏感。NIM对COX-2表达阴性的MCF-7细胞同样具有抑制增生、诱导凋亡的作用。结论 NIM对ER阳性和ER阴性乳腺癌细胞均有抑制增生、诱导凋亡的作用。NIM的抗肿瘤作用存在环氧化酶．2依赖性与非依赖性两种途径。     

The effect of non-steroidal anti-inflammatory drugs on human colorectal cancer cells: evidence of different mechanisms of action

Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit proliferation and induce apoptosis in human colorectal cancer cells in vitro. It remains unclear whether individual NSAIDs act by cyclooxygenase-2 (COX-2) inhibition and how NSAIDs exert their anti-proliferative effects. We investigated the effects of NS-398 (a selective COX-2 inhibitor), indomethacin (a non-selective COX inhibitor) and aspirin on four human colorectal cancer cell lines (HT29.Fu, HCA-7, SW480 and HCT116). NS-398 completely inhibited proliferation, induced G1 arrest and promoted apoptosis in COX-2-expressing cells (HT29.Fu and HCA-7). However, indomethacin had similar effects on all cells, regardless of COX-2 expression. NS-398 also had anti-proliferative activity on COX-2-negative cell lines (SW480 and HCT116). Aspirin inhibited proliferation of all cell lines but did not induce apoptosis. Indomethacin decreased beta-catenin protein expression in all cells (unlike NS-398 or aspirin). NSAIDs act on human colorectal cancer cells via different mechanisms. Decreased beta-catenin protein expression may mediate the anti-proliferative effects of indomethacin.     

Arginine deiminase enhances dexamethasone-induced cytotoxicity in human T-lymphoblastic leukemia CCRF-CEM cells

Since arginine deiminase (ADI; EC 3.5.3.6) inhibits cell proliferation by arresting cells in the G1 phase, we tested its synergistic effect on cell death induced by dexamethasone (DEX), which also induces apoptosis by G1 cell cycle arrest. ADI inhibited cell proliferation and induced apoptosis in human leukemic CEM cells in a dose-dependent manner. Simultaneous treatment with ADI and DEX showed synergistic effects on DNA fragmentation and LDH release. In addition, ADI exerted its anti-proliferative activity against DEX-resistant CEM cells. ADI suppressed expression of c-myc, a potential key regulator of cell proliferation and apoptosis, and increased expression of p27Kip1 cyclin-dependent kinase inhibitor. These results suggest that ADI efficiently increases the anti-cancer effect of DEX on human leukemic CEM cells through G1 cell cycle arrest involving downregulation of c-myc and upregulation of p27Kip1.     

Promising Anticancer Effect of Aurisin A Against the Human Lung Cancer A549 Cell Line

Objective: To investigate the anticancer effect of aurisin A and the underlying mechanisms of its action on the human lung cancer A549 cell line. Methods: Cell viability was determined by sulforhodamine B (SRB) assay, while cell cycle distribution and apoptosis were measured by flow cytometry. The molecular underlying mechanisms of anti-cancer properties of aurisin A was determined by western blot analysis. Results: Aurisin A exerts its anticancer effects by inhibiting cell growth (p<0.001), increasing the proportion of cells at the G0/G1 phase (p<0.001), and decreasing the expression of cyclin D (p<0.05) and cyclin-dependent kinase 4 (Cdk-4) (p<0.001). Nuclear morphological changes were observed in aurisin A-treated cells, demonstrated by a dose-dependent increase in the number of apoptosis cells (p<0.001). After aurisin A treatment, B-cell lymphoma 2 (Bcl-2) was down-regulated (p<0.05), cleaved caspase-3 was up-regulated (p<0.05). In addition, aurisin A inhibits migration of cancer cells in a dose-dependent manner (p<0.001) and decreases the expression of epidermal growth factor receptor (EGFR) (p<0.05) and phosphorylated p38 (pp38) (p<0.05). Conclusion: These results indicated that in-vitro treatment of aurisin A against this human lung cancer cell line inhibits cell proliferation and migration, and induces apoptosis and cell-cycle arrest.  Aurisin A is a promising anticancer agent for use against human lung cancer.     

Silibinin Inhibits Proliferation,Induces Apoptosis and Causes Cell Cycle Arrest in Human Gastric Cancer MGC803 Cells Via STAT3 Pathway Inhibition

Background: To investigate the effect of silibinin on proliferation and apoptosis in human gastric cancer cell line MGC803 and its possible mechanisms. Materials and Methods: Human gastric cancer cell line MGC803 cells were treated with various concentration of silibinin. Cellular viability was assessed by CCK-8 assayandapoptosis and cell cycle distribution by flow cytometry. Protein expression and mRNA of STAT3, and cell cycle and apoptosis regulated genes were detected by Western blotting and real-time polymerase chain reaction, respectively. Results: Silibinin inhibits growth of MGC803 cells in a dose- and time-dependent manner. Silibinin effectively induces apoptosis of MGC803 cells and arrests MGC803 cells in the G2/M phase of the cell cycle, while decreasing the protein expression of p-STAT3, and of STAT3 downstream target genes including Mcl-1, Bcl-xL, survivin at both protein and mRNA levels. In addition, silibinin caused an increase in caspase 3 and caspase 9 protein as well as mRNA levels. Silibinin caused G2/M phage arrest accompanied by a decrease inCDK1 and Cyclin B1 at protein and mRNA levels.. Conclusions: These results suggest that silibinin inhibits the proliferation of MGC803 cells, and it induces apoptosis and causes cell cycle arrest by down-regulating CDK1, cyclinB1, survivin, Bcl-xl, Mcl-1 and activating caspase 3 and caspase 9, potentially via the STAT3 pathway.     

蛴螬粗提物对人宫颈癌HeLa细胞诱导凋亡作用及其机制

 目的 研究蛴螬粗提物对人宫颈癌HeLa细胞诱导凋亡作用，并探讨其作用机制。方法 采用流式细胞仪检测HeLa细胞的细胞周期分布相和细胞凋亡率；末端脱氧核苷酸转移酶（TdT）介导的脱氧核苷酸缺口末端标记（TdT mediated dUTP nick end labeling）TUNEL法检测凋亡细胞；通过免疫细胞化学SP法测定凋亡相关基因产物p53、Fas、bcl 2、Bax蛋白的表达情况。结果 (1)蛴螬粗提物可以诱导HeLa细胞发生凋亡，加药组出现亚二倍体峰，并且G0/G1期细胞比例显著增加（P＜0.01），而S期和G2/M期细胞比例则明显下降,将细胞阻滞在G0/G1期；（2）蛴螬粗提物作用细胞后凋亡细胞数目随时间延长而增多，凋亡指数与药物处理呈时间依赖性和剂量依赖性；(3)蛴螬粗提物作用后bcl 2、p53蛋白随提取物浓度表达均下降，Bax、Fas蛋白表达上升，四种蛋白表达各组间比较差异均有具有统计学意义（P<0.01）。结论 (1) 蛴螬粗提物对人宫颈癌HeLa细胞具有诱导凋亡作用；(2)蛴螬粗提取物诱导凋亡作用机制可能与细胞周期发生G0/G1期阻滞有关；并且通过下调bcl 2、p53蛋白表达,上调Bax、Fas蛋白表达，经由细胞凋亡的死亡受体通路和线粒体通路完成凋亡的启动和执行。     

盐酸小檗碱对人胃癌细胞增殖和凋亡的影响

目的：研究盐酸小檗碱对人胃癌细胞及永生化胃上皮细胞增殖和凋亡的影响,初步探讨其中可能的分子机制。方法：利用不同浓度的小檗碱处理胃癌细胞系MKN-45和SGC-7901细胞,以及永生化胃上皮细胞系GES细胞,采用MTT法测定细胞活力,流式细胞术检测细胞周期和细胞凋亡（PI标记）,Western blot检测细胞周期和凋亡相关蛋白的表达。结果：MTT结果显示与空白对照组相比,小檗碱处理后,细胞增殖速度明显受到抑制（P〈0.05）,并呈浓度依赖性,其中50μg/ml处理组最大抑制率为81.3%,这种抑制作用对胃癌细胞更为明显。流式细胞仪检测表明,小檗碱在较低浓度（10μg/ml）时即可诱导胃癌细胞出现G0/G1阻滞,S期细胞百分比减少和细胞凋亡（P〈0.05）,这种作用呈时间和浓度依赖性。Western blot结果提示小檗碱处理后,胃癌细胞Bcl-2蛋白表达水平降低,Bax及p53蛋白表达水平明显上调。结论：小檗碱可抑制胃癌细胞增殖,诱导胃癌细胞凋亡,其机制与阻滞细胞周期进展以及调控凋亡相关蛋白Bcl-2、Bax和p53密切相关。     

G-CSF在人实体肿瘤细胞株中的表达及其对细胞增殖的影响

目的:探讨G-CSF在人实体肿瘤细胞株中的表达及外源性rhG-CSF对肿瘤细胞增殖的影响及机制。方法:人G-CSF ELISA试剂盒检测肿瘤细胞G-CSF的分泌水平,细胞计数法和MTT法检测rhG-CSF对肿瘤细胞增殖的影响,流式细胞仪分析rhG-CSF对KB细胞周期和细胞凋亡的影响,免疫细胞化学检测G-CSF受体在肿瘤细胞中的表达。结果:T-24细胞分泌大量G-CSF;rhG-CSF呈剂量依赖性促KB细胞增殖,并在质量浓度为100 ng/mL达最大值,与对照相比,差异有统计学意义(P<0.05);KB细胞表达G-CSF受体,rhG-CSF拮抗KB细胞的G0/G1期阻滞,降低KB细胞凋亡率。结论:T-24细胞表达G-CSF;rhG-CSF通过作用于KB细胞表面的G-CSF受体拮抗G0/G1期阻滞、抑制凋亡而发挥促增殖作用。     

HIF-1α抑制剂YC-1对人骨肉瘤细胞Saos2和U2-OS增殖和凋亡的影响

目的:应用缺氧诱导因子-1α（hypoxia inducible factor-1α,HIF-1α）抑制剂YC-1处理人骨肉瘤细胞Saos2和U2-OS，观察不同药物浓度对人骨肉瘤细胞系Saos2和U2-OS细胞增殖、细胞周期以及凋亡的影响及其机制。方法:人骨肉瘤细胞系Saos2和U2-OS分对照组和实验组。对照组细胞1%氧气浓度低氧培养，实验组细胞施加HIF-1α抑制剂YC-1干预，设浓度梯度。CCK-8法检测不同药物浓度对Saos2和U2-OS细胞活力及增殖的影响；流式细胞术检测YC-1对Saos2和U2-OS细胞周期和凋亡的影响；Western blot对FXR1、G3BP1、APAF1、Cleaved Caspase-3蛋白的表达进行相对定量分析。结果:CCK-8检测结果显示，YC-1对人骨肉瘤Saos2和U2-OS细胞有显著的增殖抑制作用，且呈明显的剂量依赖效应。不同浓度的YC-1处理细胞后，均可使各处理组G1期细胞增多，S期细胞减少，且呈剂量依赖效应(P＜0.05)。不同浓度的YC-1处理细胞后，与对照组相比，各实验组FXR1、G3BP1、APAF1蛋白表达显著下降(P＜0.05)，而Cleaved Caspase-3蛋白表达显著增高(P＜0.05)。结论:YC-1可以将人骨肉瘤细胞株Saos2和U2-OS细胞阻滞于G1期，影响细胞周期进程，抑制细胞增殖，促进细胞凋亡。HIF-1α对人骨肉瘤细胞增殖与凋亡的影响可能是通过上调FXR1、G3BP1、APAF1的表达而实现，而YC-1抑制了HIF-1α的表达从而使Cleaved Caspase-3的表达增高诱导细胞凋亡。HIF-1α及FXR1、G3BP1、APAF1、Cleaved Caspase-3有望成为骨肉瘤治疗的新靶标。     

Histone deacetylase inhibitors such as sodium butyrate and trichostatin A induce apoptosis through an increase of the bcl-2-related protein Bad

The effects of sodium butyrate (SB) and trichostatin A (TSA) on cell proliferation and apoptosis against human glioma T98G, U251MG, and U87MG cells were investigated. Upon exposure to either SB or TSA, cell proliferation was reduced, and apoptosis detected by DNA fragmentation analysis and the cleavage of CPP32 was induced. Previously, we reported that SB increased the expression levels of p21 (WAF-1) and inhibited G1-S transition of the cell cycle. In this study, we showed that TSA also increased p21 expression, suggesting that histone deacetylase (HDAC) inhibitors may up-regulate p21 protein in common and thus arrest proliferation in the G1 phase of the cell cycle. To further determine the underlying molecular mechanisms of apoptosis with either SB or TSA treatment, we studied the expression levels of apoptosisrelated proteins in human glioma cells. SB increased the expression of the Bad protein, although the expression of Bcl-2, Bcl-xL, Bax, and Fas was not changed by the addition of SB. TSA treatment also up-regulated the expression of Bad protein. The results suggest that HDAC inhibitors such as SB and TSA induce apoptosis through an increase in Bad protein in human glioma cells in vitro.