^131I-GMCSF诱导HL-60细胞凋亡与细胞周期阻滞的研究

目的观察和探讨131Ⅰ-GMCSF诱导HL-60细胞凋亡及其与细胞周期的关系.方法采用体外放射免疫治疗模型,通过MTT比色法、TUNEL来研究131Ⅰ-GMCSF辐射后HL-60细胞的存活率、凋亡率的改变;用流式细胞术及免疫细胞化学方法细胞周期的改变.结果放射性浓度≥18.5×108 Bq/L时HL-60细胞存活率显著下降.131Ⅰ-GMCSF能致HL-60细胞凋亡、细胞发生G2/M期阻滞.结论 GMCSF作为载体携带131Ⅰ能诱导HL-60细胞凋亡,凋亡与细胞发生G2/M期阻滞有关.     

131Ⅰ-GMCSF诱导HL-60细胞凋亡与细胞周期阻滞的研究

目的观察和探讨131Ⅰ-GMCSF诱导HL-60细胞凋亡及其与细胞周期的关系.方法采用体外放射免疫治疗模型,通过MTT比色法、TUNEL来研究131Ⅰ-GMCSF辐射后HL-60细胞的存活率、凋亡率的改变;用流式细胞术及免疫细胞化学方法细胞周期的改变.结果放射性浓度≥18.5×108 Bq/L时HL-60细胞存活率显著下降.131Ⅰ-GMCSF能致HL-60细胞凋亡、细胞发生G2/M期阻滞.结论 GMCSF作为载体携带131Ⅰ能诱导HL-60细胞凋亡,凋亡与细胞发生G2/M期阻滞有关.     

~(131)I-GMCSF诱导HL-60细胞凋亡与细胞周期阻滞的研究

目的　观察和探讨1 31 I-GMCSF诱导HL - 6 0细胞凋亡及其与细胞周期的关系。方法　采用体外放射免疫治疗模型,通过MTT比色法、TUNEL来研究1 31 I -GMCSF辐射后HL - 6 0细胞的存活率、凋亡率的改变;用流式细胞术及免疫细胞化学方法细胞周期的改变。结果　放射性浓度≥18.5×10 8Bq/L时HL - 6 0细胞存活率显著下降。1 31 I -GMCSF能致HL - 6 0细胞凋亡、细胞发生G2 /M期阻滞。结论　GMCSF作为载体携带1 31 I能诱导HL - 6 0细胞凋亡,凋亡与细胞发生G2 /M期阻滞有关。     

土贝母皂苷诱导人宫颈癌HeLa细胞周期阻滞和细胞凋亡

背景和目的：土贝母皂苷是由传统中药土贝母块茎中分离得到的,由土贝母苷甲（79%）和土贝母苷乙（21%）组成。本研究旨在探讨土贝母皂苷抗肿瘤作用的机制。方法：MTT法检测土贝母皂苷对肿瘤细胞生长的抑制作用,流式细胞仪分析细胞周期,荧光显微镜和电子显微镜观察细胞形态,琼脂糖凝胶电泳检测DNA电泳图谱的变化。结果：土贝母皂苷对人宫颈癌HeLa细胞的生长具有强制作用,且这种作用呈时间剂量依赖关系,处理24、48和72h的IC50值分别为20.0、18.8和8.8μmol/L.流式细胞术分析显示,15、30、35μmol/L土贝母皂苷处理5h,HeLa细胞G2/M期细胞数从9.80%分别增加到21.90%、25.92%和27.00%;处理12h,G2/M期细胞数增加更显著,从8.20%分别增加到21.40%、31.15%和34.55%。40μmol/L土贝母皂苷处理一定时间,形态学检查发现细胞核明显皱缩、核染色质凝集边聚等特征,流式细胞术检测发现凋亡峰,DNA琼脂糖凝胶电泳可见明显的梯形条带。结论：土贝母皂苷使细胞周期阻滞和诱导细胞凋亡可能在其抗肿瘤作用中具有重要意义。     

细胞色素C诱导HL-60细胞凋亡及其周期特异性分析

目的分析细胞色素C诱导HL-60细胞凋亡的时间效应及其细胞周期的特异性.方法 (1)利用透射电镜观察细胞色素C诱导HL-60细胞凋亡时凋亡小体的形成;(2)应用流式细胞仪技术检测细胞色素C诱导HL-60细胞的凋亡率并分析其凋亡的时-效关系以及凋亡与细胞周期的关系.结果 (1)细胞色素C作用HL-60细胞24小时发生凋亡,形成凋亡小体;(2)细胞色素C诱导HL-60细胞凋亡的作用体现一定的时间效应,作用时间不同,诱导凋亡率亦不相同,其中作用24小时凋亡率最高;(3)细胞色素C诱导HL-60凋亡的作用具有细胞周期特异性,随凋亡率升高,S期和G2期细胞比例随之下降,且S期细胞比例与凋亡率二者呈显著负性关系(r=0.718,P＜0.01).结论细胞色素C诱导HL-60细胞凋亡与细胞周期密切相关,其周期特异性具有一定的实际意义.     

鲎血细胞多肽诱导HL-60细胞凋亡

目的：研究鲎血细胞多肽（下称鲎血多肽）诱导HL－60细胞凋亡的作用。方法：MTT法测定鲎血多肽对HL－60细胞的毒性和HL－60细胞的相对存活率,荧光显微镜观察HL－60细胞形态的变化,流式细胞仪鉴定细胞凋亡和分析细胞周期,扫描电镜观察细胞膜的改变。结果：鲎鱼多肽对HL－60细胞有明显的细胞毒性,IC50值为24μg/ml;荧光显微镜观察到凋亡细胞主要表现为细胞体积缩小,核染色质固缩,荧光染色增强。50－100μg/ml的鲎血多肽处理6h,HL－60细胞主要表现为细胞凋亡;而鲎血多肽浓度为200μg/ml时的主要为细胞坏死。50μg/ml鲎血多肽处理HL－60细胞0－12h,流式细胞仪检测到凋亡细胞亚二倍体峰出现,细胞周期分析G1期细胞减少,G2期细胞增加。鲎血多肽浓度为25－100μg/ml时,细胞凋亡率随浓度增加而增加,鲎血多肽为200μg/ml时,则主要为细胞坏死,凋亡细胞减少。扫描电镜观察发现,HL－60细胞用鲎血多肽处理后,细胞膜损伤,出现空洞。结论：鲎血细胞多肽能诱导HL－60细胞凋亡;凋亡的发生较早,与细胞膜受损有一定的关系;G1期细胞对鲎血多肽更敏感。     

雷公藤多甙诱导HL-60细胞凋亡

目的：探讨雷公藤多甙对ＨＬ－６０细胞凋亡的作用。方法：应用细胞形态学检查,ＤＮＡ凝胶电泳及流式细胞仪分析检测。结果：ＨＬ－６０细胞加上雷公藤多甙１０ｍｇ／Ｌ孵育７２ｈ,流式细胞仪检测细胞凋亡率为２９．４％（与空白对照组比较,Ｐ〈０．０１）。电检查和光镜检查均可见细胞核固缩、碎裂等。ＤＮＡ电泳显示明显的梯状条带。结论：雷公藤多甙能诱导ＨＬ－６０细胞凋亡,提示雷公藤多甙可能具有抗白血病作用。     

宫颈癌细胞周期阻滞和凋亡与放射敏感性的关系

宫颈癌是我国女性最常见的恶性肿瘤之一。放射治疗是治疗宫颈癌的主要方法 ,但疗效仍不满意。如果在放射治疗前即可预测肿瘤的放射敏感性 ,制定个体化的治疗方案 ,则可能提高疗效。笔者搜集了 45例宫颈癌病例 ,对其癌组织标本细胞周期及凋亡在放射治疗过程中的变化与放射敏感性的关系进行了研究 ,现报道如下。一、材料和方法1.临床资料 :1999年 12月至 2 0 0 1年 5月在本院就诊、未经治疗的宫颈鳞癌患者 45例 ,诊断均经病理学检查证实。中位年龄 48岁 (2 9～ 80岁 )。国际妇产科协会 (ＦＩＧＯ)临床分期ＩＩＢ期 4例 (8.89% ) ,ⅢＢ期 41例 …     

丹参酮ⅡA诱导HL-60细胞凋亡

目的：在以往对丹参酮ＩＡ诱导分化和抗癌作用研究的基础上，进一步研究丹参酮ＩＡ诱导ＨＬ－６０细胞凋亡，以探讨其抗癌作用机理。方法：应用体外细胞培养技术，以１～１０μｇ／ｍｌ丹参酮ＩＡ作用于培养的ＨＬ－６０细胞５天后，进行光镜、电镜、琼脂糖凝胶电泳及流式细胞仪的观察和分析。结果：丹参酮ⅡＡ作用后，ＨＬ－６０细胞的生长受到明显抑制，其半数抑制浓度ＩＣ５０约为１０μｇ／ｍｌ光镜、电镜观察到细胞呈现凋亡特征，琼脂糖凝胶电泳显示细胞ＤＮＡ呈梯状降解，流式细胞仪分析表明，１０μｇ／ｍｌ丹参酮ⅡＡ作用后，ＨＬ－６０细胞凋亡率达２８．８％，细胞被阻止于Ｇ０／Ｇ１期，Ｓ期细胞明显减少（Ｐ＜０．０１），其ｂｃｌ－２基因的表达显著降低，而ｐ５３基因的表达增强（Ｐ＜０．０１）。结论：丹参酮ⅡＡ可诱导ＨＬ－６０细胞凋亡，这可能为丹参酮ⅡＡ抗癌抑癌的重要机理之一，诱导分化与诱导凋亡之间的关系有待进一步探讨。     

羟基喜树碱诱导HL-60细胞凋亡的研究

目的 :探讨国产羟基喜树碱 (HCPT)对 HL - 6 0细胞的作用机制和规律。方法 :应用细胞形态学检查 ,DNA凝胶电泳及流式细胞仪分析检测。结果 :HCPT 1mg/ L和 5 mg/ L作为 HL - 6 0细胞 4h细胞凋亡率分别为 5 0 .6 %和 6 8.6 %。光镜检查见细胞核固缩、碎裂 ;电镜检查见染色质向核周边凝集。 DNA电泳显示典型的梯状条带。当 HCPT浓度超过 5 mg/ L或作用超过 4h,细胞凋亡率无明显增加 ,出现饱和现象。HL - 6 0细胞经 HCPT作用后 ,S期细胞显著减少 ,G0 / G1 期细胞增加。结论 :HCPT诱导 HL - 6 0细胞出现凋亡并具有饱和效应 ,HCPT为 S期特异性药物。临床上 HCPT应与细胞周期非特异性药物联合用药。     

Quercetin Induces Cell Cycle Arrest and Apoptosis in YD10B and YD38 Oral Squamous Cell Carcinoma Cells

Objective: Oral squamous cell carcinoma (OSCC) exhibits the highest lethality among head and neck cancers. Treatment for OSCC is limited due to diverse side effects. Quercetin is a natural flavonoid compound found in many kinds of plants and foods. Quercetin has been reported to be a modulator of proliferation and survival in various types of cancers due to its cytotoxic effects. We aimed to investigating chemopreventative roles of quercetin in YD10B and YD38 OSCC cells. Methods: For our study, two different types of OSCC cells were used. YD10B cells are tongue SCC cells with the p53 mutation and YD38 cells are lower gingiva SCC cells without the p53 mutation, respectively. The anticancer effects of quercetin were examined by cell viability, cell cycle, annexin-PI staining, and western blot. Result: Our results showed that quercetin decreased cell viability and induced G1 cell cycle arrest in YD10B and YD38 OSCC cells. Moreover, quercetin remarkably decreased the expression of cell cycle upregulating proteins and increased the expression of a CDK inhibitor. Quercetin also significantly increased the number of annexin-V-positive cells in a dose-dependent manner in both types of OSCC cells. This apoptotic potential of quercetin triggered cleavage of PARP followed by activation of p38 MAPK signaling pathway. Conclusion: In conclusion, this study demonstrates that quercetin shows different anti-cancer responses in OSCC with and without p53 mutation, respectively. Despite different p53 status in OSCC cells, quercetin led to apoptotic signals in both cells. Quercetin repressed cell proliferation with G1 cell cycle arrest and apoptosis by activating the p38 signaling pathway in two OSCC cells with different p53 status. These findings might provide new strategy for OSCC therapy by quercetin.     

土贝母苷甲对人HL-60髓性白血病细胞周期与凋亡的影响

目的:研究土贝母苷甲(下称苷甲)对人髓性白血病细胞HL-60的细胞周期及凋亡的影响。方法:采用MTT(四甲基偶氮唑蓝)法检测苷甲对HL-60细胞生长的影响;形态学方法(荧光显微镜和透射电镜)、流式细胞仪和DNA琼脂糖凝胶电泳观察和分析在苷甲作用下HL-60细胞形态、DNA含量的变化和DNA断裂的情况;蛋白印迹免疫法检测细胞凋亡及周期相关基因bcl-2、cyclinB1表达的变化。结果:苷甲显著抑制HL-60细胞的生长,其抑制效果与浓度及时间呈依赖关系。15μmol·L-1苷甲作用24小时可将HL-60细胞阻滞于G2/M期,进而诱导其凋亡,凋亡细胞具有典型的凋亡形态特征,琼脂糖凝胶电泳可见明显的梯状区带。bcl-2表达无明显变化,cyclinB1表达降低。结论:苷甲对细胞周期起阻滞作用,并诱导其凋亡,其作用机制可能与cyclinB1表达降低有关。     

Bracken-fern Extracts Induce Cell Cycle Arrest and Apoptosis in Certain Cancer Cell Lines

Bracken fern [Pteridium aquilinem (L.) kuhn (Dennstaedtiaceae)] is one of the most common species on the planet. It has been consumed by humans and animals for centuries. Use by some human groups is because theybelieve bracken fern is good for health as plant medicine. However, it is also one of the few known plants that can cause tumors in farm animals. Many interested groups have focused their attention on bracken fern because of these interesting features. In order to evaluate the biological effects of exposure to this plant in cellular level, human cancer cell lines were treated with the fern dichloromethane extracts and the genotoxic and cytotoxic effects were studied. Anti-proliferative/cytotoxic effects were evaluated by cell count, MTT assay and flow cytometry methods with three different cancer cell lines, TCC, NTERA2, and MCF-7, and two normal cells, HDF1 and HFF3. Pro-apoptotic effects of the extracts were determined by DAPI staining and comet assay, on TCC cancercells compared to the normal control cell lines. Cellular morphology was examined by light microscopy. Our present study showed that the extract caused DNA damage and apoptosis at high concentrations (200 μg/mL)and also it may induce cell cycle arrest (G2/M phase) at mild concentrations (50 and 30 μg/mL) depending on the cell type and tumor origin. These results indicate that bracken fern extract is a potent source of anticancercompounds that could be utilized pharmaceutically.     

Cytotoxicity,Cell Cycle Arrest,and Apoptosis Induction Activity of Ethyl-p-methoxycinnamate in Cholangiocarcinoma Cell

Objective: To investigate cytotoxic activity of ethyl-p-methoxycinnamate (EPMC) including its effect on p-glycoprotein (multidrug resistance-1: mdr-1 gene) in human cholangiocarcinoma cell. Methods: Cytotoxic activity of EPMC against human cholangiocarcinoma (CL-6), fibroblast (OUMS-36T-1F), and colon cancer (Caco-2) cell lines were assessed using MTT assay. Selectivity index (SI) was determined as the ratio of IC50 (concentration that inhibits cell growth by 50%) of EPMC in OUMS-36T-1F and that in CL-6 cell. Cell cycle arrest and apoptosis in CL-6 cells were investigated by flow cytometry and fluorescent microscopy. Effect of EPMC on mdr-1 gene expression in CL-6 and Caco-2 was determined by real-time PCR. Results: The median (95% CI) IC50 values of EPMC in CL-6, OUMS-36T-1F, and Caco-2 were 245.5 (243.1-266.7), 899.60 (855.8-966.3) and 347.0 (340.3-356.9) µg/ml, respectively. The SI value of the compound for the CL-6 cell was 3.70. EPMC at IC50 inhibited CL-6 cell division and induced apoptosis compared to untreated control. EPMC exposure did not induce mdr-1 gene expression in both CL-6 and Caco-2 cells. Conclusion: The results suggest the potential role of EPMC in cholangiocarcinoma with a low possibility of drug resistance induction.     

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.     

Xanthoxyletin,a Coumarin Induces S Phase Arrest and Apoptosis in Human Gastric Adenocarcinoma SGC-7901 Cells

This study was conducted to explore the novel anticancer compounds from Chinese herbs. During the processof screening, to evaluate the potential chemopreventive effect of natural compounds, Xanthoxyletin was isolatedfrom Erythrina variegata. It has been reported that Xanthoxyletin possesses antibacterial, fungicidal, and algicidalproperties. In this study, we examined the antiproliferative effects of Xanthoxyletin against SGC-7901 cells andits ability to induce apoptosis and cell cycle arrest for the first time. We observed that its inhibitory effects oncells were associated with the DNA damage, apoptosis through mitochondrial dysfunction, and cell cycle arrestat S phase in a dose-dependent manner. Additionally, Xanthoxyletin also increased the production of reactiveoxygen species in SGC-7901 cells. These results suggest that Xanthoxyletin may be promising anticancer agentand has worth for further mechanistic and therapeutic studies against gastric cancer.     

Coordinate Involvement of Cell Cycle Arrest and Apoptosis Strengthen the Effect of FTY720

A novel reagent, FTY720 (2-amino-2-[2-(4-octylphenyl)ethyl]-l,3-propanediol hydrochloride), has been shown to induce a significant decrease of lymphocytes and lymphoma cells and is expected to be a potent immunosuppressant and anti-tumor drug. The decrease in lymphocytes and lymphoma cells is mainly the result of FTY720-induced apoptosis. FTY720 directly affects mitochondria and induces cell death. Moreover, FTY720 activates protein phosphatase (PP) 2A and affects anti-apo-ptotic intracellular signal transduction proteins to attenuate the anti-apoptotic effect. In this study, we examined the relationship between FTY720-induced apoptosis and cell cycle regulation. FTY720 induced apoptosis significantly at the GO/G1 phase and caused GO/G1 cell cycle arrest of the human lymphoma cell lines HL-60RG and Jurkat. Simultaneously, retinoblastoma protein (pRB) was dephosphorylated, suggesting that dephosphorylation of pRB was related to FTY720-induced GO/G1 cell cycle arrest. Because this dephosphorylation was completely blocked by a specific PP1/ 2A inhibitor, okadaic acid, it appears that FTY720-activated PP2A is essential for FTY720-induced cell cycle arrest. FTY720-induced apoptosis was inhibited by Bcl-2 overexpression in Jurkat cells, but this did not prevent FTY720-induced cell cycle arrest, suggesting that the mechanism of FTY720-induced cell cycle arrest is independent of the mechanism of FTY720-induced apoptosis. These two independent pathways strengthen the effect of FTY720.     

Iso-suillin Isolated from Suillus luteus,Induces G1 Phase Arrest and Apoptosis in Human Hepatoma SMMC-7721 Cells

Iso-suillin, a natural product isolated from Suillus luteus, has been shown to inhibit the growth of somecancer cell lines. However, the molecular mechanisms of action of this compound are poorly understood. Thepurpose of this study was to investigate how iso-suillin inhibits proliferation and induces apoptosis in a humanhepatoma cell line (SMMC-7721). We demonstrated the effects of iso-suillin on cell proliferation and apoptosisin SMMC-7721 cells, with no apparent toxicity in normal human lymphocytes, using colony formation assaysand 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Western blotting was usedto examine the expression of G1 phase-regulated and apoptosis-associated protein levels in iso-suillin treatedSMMC-7721 cells. The results indicated that iso-suillin significantly decreased viability, induced G1 phase arrestand triggered apoptosis in SMMC-7721cells. Taken together, these results suggest the potential of iso-suillin asa candidate for liver cancer treatment.     

新合成吩嗪衍生物体外诱导癌细胞周期阻滞和凋亡的实验

目的 探讨新合成吩嗪衍生物抗癌活性及其作用机制。方法 MTT法检测细胞增殖,吖啶橙/溴乙锭（AO/EB）染色荧光显微镜观察细胞形态,流式细胞仪检测细胞周期,Annexin V-FITC/7-AAD双染检测凋亡,Western blot检测p53和caspase-3蛋白的表达。结果 pn18、pn23、pc27和pc28四种化合物在体外抑制癌细胞增殖,尤其对人结直肠癌细胞HCT116作用明显,且呈时间和剂量依赖性。pc28在四种化合物中作用最强,48 h对HepG2、HCT116和A549细胞半数抑制浓度（IC₅₀）分别为（6.62±2.69）、（10.83±1.41）和（22.39±4.31）μmol。20 μmol化合物作用于HCT116细胞24 h后,细胞密度降低,形态变圆,细胞内明亮绿色荧光与染色质浓缩相关,死亡细胞呈橙黄色和红色与细胞膜通透性增加有关;pc28诱导的细胞晚期凋亡比例由0.345%升高至19.7%。20 μmol pn18和pc27诱导G₀/G₁期细胞分别升高17.5%和25.0%,pn23和pc28诱导S期细胞分别升高18.4%和11.0%。pn23和pc28诱导p53表达升高,pc28也能上调并激活caspase-3。结论 四种合成的新型吩嗪衍生物在体外能有效抑制多种癌细胞增殖,其作用机制可能与诱导细胞周期阻滞和凋亡相关。