三氧化二砷诱导卵巢癌OVCAR-3细胞周期阻滞及凋亡

目的探讨As2O3对卵巢癌细胞增殖的抑制作用及对细胞周期和凋亡的影响.方法采用MTT法及集落形成实验测定细胞的增殖活力,通过细胞形态学观察细胞分裂及凋亡,应用流式细胞仪进行细胞周期解析、凋亡及bcl-2蛋白表达的检测.结果As2O3呈剂量依赖性抑制OVCAR-3细胞增殖,抑制50%细胞生长的药物浓度(IC50)为2μmol/L.0.5～5μmol/L的As2O3作用7天,集落抑制率均在40%以上(P＜0.01).2μmol/L与5μmol/L的As2O3作用12 h后,细胞周期出现了G2/M期阻滞及亚二倍体凋亡峰,随着作用时间的延长,凋亡细胞随G2/M期细胞减少而逐渐增多.细胞形态学观察可见分裂期细胞及凋亡细胞明显增加.0.5～5μmol/L的As2O3均能下调bcl-2蛋白的表达.结论As2O3能够抑制卵巢癌OVCAR-3细胞的增殖,诱导M期阻滞及细胞凋亡.     

奥沙利铂联合替尼泊苷抑制胃癌BGC-823细胞生长和诱导凋亡的作用

目的:探讨奥沙利铂(oxaliplatin, OXA)联合替尼泊苷(teniposide, VM-26)抑制胃癌BGC-823细胞的体外生长及诱导细胞凋亡的作用.方法:应用MTT法检测不同浓度OXA和VM-26单药以及联合用药对胃癌BGC-823细胞生长的抑制作用,应用FCM检测细胞凋亡率,同时应用免疫细胞化学法检测OXA单药以及OXA联合VM-26用药对胃癌BGC-823细胞caspase-9和livin蛋白表达的影响.结果:OXA和VM-26单药均可抑制胃癌BGC-823细胞的生长,且在一定浓度范围内呈剂量依赖效应;联合用药组的细胞生长抑制率明显高于OXA和VM-26单药组,差异有统计学意义(P<0.05),联合指数(combination index, CI)为0.46.OXA(1.250 μg/mL)单药作用于胃癌BGC-823细胞12、24和48 h后,细胞凋亡率分别为6.13%、13.86%和21.48%;VM-26(0.625 μg/mL)单药组的细胞凋亡率分别为4.60%、10.72%和17.07%;联合用药组的细胞凋亡率分别为11.73%、24.14%和44.75%.联合用药组的细胞凋亡率明显高于OXA和VM-26单药组,差异有统计学意义(P<0.05).免疫细胞化学结果显示,OXA(1.250 μg/mL)单用以及联合VM-26(0.750 μg/mL)作用于胃癌BGC-823细胞48 h后,凋亡相关基因caspase-9蛋白的阳性表达率均增加,而livin蛋白的阳性表达率均下降;各用药组之间以及用药组与对照组(未用药)之间的差异均有统计学意义(P<0.05).结论:OXA联合VM-26用药在抑制胃癌BGC-823细胞的生长和诱导细胞凋亡方面具有协同作用.     

Coleusin factor exerts cytotoxic activity by inducing G0/G1 cell cycle arrest and apoptosis in human gastric cancer BGC-823 cells

Coleusin factor (CF), a kind of diterpenoids, is isolated and purified from the root of a Chinese tropical plant Coleus forskohlii by our laboratory. Our previous studies have demonstrated that CF significantly inhibits growth in some kinds of cancer cell lines. Here, we found that CF remarkably inhibited growth in human gastric cancer BGC-823 cells by decreasing cell proliferation, inducing G(0)/G(1) cell cycle arrest and apoptosis. CF also decreased the mitochondrial membrane potential in BGC-823 cells. Immunoblotting analysis revealed that CF significantly decreased the expressions of cyclinD1, Bcl-2, and Bcl-x(L), increased the expressions of cytosol cytochrome c, p53, p21, and Rb. In addition, CF significantly increased the expressions and activities of caspase-3 and -9. More importantly, CF potently inhibited the growth of BGC-823 cells xenografted in athymic nude mice with negligible body weight loss and damage towards the spleen. These results indicate that CF exerts a cytotoxic effect on BGC-823 cells by inducing cell cycle arrest and apoptosis.     

表没食子儿茶素没食子酸酯诱导人胃癌BGC-823细胞凋亡的机制

目的:探讨表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对人胃癌BGC-823细胞增殖的影响及其可能的机制。方法:不同浓度EGCG单独或联合c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)抑制剂SP600125作用BGC-823细胞后,应用MTT法检测BGC-823细胞的增殖抑制率,显微镜下观察细胞的形态学改变,FCM检测细胞凋亡率,蛋白质印迹法检测细胞中p-JNK和p-c-jun蛋白的表达情况,免疫细胞化学法检测细胞中caspase-3的表达。结果:EGCG可抑制BGC-823细胞的增殖(P<0.05),呈时间和剂量依赖效应;EGCG可诱导BGC-823细胞凋亡,细胞凋亡率呈剂量依赖效应(P<0.05);EGCG可上调BGC-823细胞中p-JNK、p-c-jun和caspase-3的表达(P<0.05)。EGCG引起的BGC-823细胞增殖抑制和caspase-3表达水平的上调可被SP600125部分抑制。结论:EGCG可诱导人胃癌BGC-823细胞凋亡,其作用机制可能与激活JNK信号转导途径并上调caspase-3的表达有关。     

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。     

HIV-1Vpr体外诱导肿瘤细胞凋亡与细胞周期阻滞的研究

目的 研究HIV-1 Vpr诱导Hela、Lovo、HepG2细胞凋亡和G2期阻滞效果.方法 构建含有HIV-1 Vpr基因的pcDNA4-Vpr重组载体,体外转染Hela、Lovo、HepG2细胞,同时设pcDNA4-EGFP质粒转染对照组、FUGENE组和空白对照组.转染后24、48、72 h,通过甲臜化合物法检测...     

奥曲肽对人胃癌BGC-823细胞增殖和凋亡影响的研究

目的:研究生长抑素类似物奥曲肽(octreotide,OCT)对胃癌BGC-823细胞增殖、凋亡、细胞周期的影响及其可能的作用机制.方法:不同浓度的OCT作用于体外培养的人胃癌BGC-823细胞,以5-FU作为阳性对照,应用MTT法分析细胞增殖的抑制作用,流式细胞仪测定细胞周期分布和凋亡率,AO-EB染色观察细胞形态变化.结果:OCT对BGC-823细胞的生长、增殖产生抑制作用,该抑制作用具有量效、时效关系和饱和性;OCT作用后细胞呈典型的凋亡形态学改变,流式细胞仪检测的细胞凋亡率和AO-EB染色测得细胞凋亡率与对照组相比较差异有显著性(P<0.05);BGC-823细胞经OCT作用后,G1期细胞数明显增加,S细胞数明显减少,细胞被阻滞于G1/S 期.结论:10-5-10-3g/L 浓度的OCT能够抑制胃癌BGC-823细胞增殖,机制可能与其诱导肿瘤细胞的凋亡和出现G1/S期阻滞有关.     

克霉唑诱导人结肠癌细胞CCL229凋亡的实验研究

背景与目的：国外有研究表明克霉唑可通过持续耗竭细胞内钙库来抑制肿瘤细胞生长,本研究旨在研究克霉唑诱导人结肠癌细胞凋亡的作用。方法：将不同浓度的克霉唑作用于体外培养的人结肠癌CCL229细胞。利用荧光显微镜,电子显微镜观察细胞的形态变化,流式细胞仪检测细胞周期变化及亚二倍体比例,并利用TUNEL法进行凋亡的分子生物学检测。结果：25－35μmol/L的克霉唑均可诱导CCL229细胞凋亡,其诱导作用表现为时间和剂量依赖性,荧光显微镜及电镜下观察到凋亡小体形成等典型的形态学改变,DNA直方图上显示亚二倍体峰,亚二倍体比例随剂量及时间的增加而增加,相应地,G1期细胞比例下降,TUNEL法观察到棕褐色着染的凋亡阳性细胞。结论：克霉唑对人结肠癌细胞系CCL229有明显的凋亡诱导作用,并且能特异地诱导G1期细胞凋亡。     

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。     

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.     

Cranberry phytochemical extracts induce cell cycle arrest and apoptosis in human MCF-7 breast cancer cells

Breast cancer is the most commonly diagnosed cancer in women in the US and is one of the leading causes of death due to cancer. Epidemiological studies have consistently suggested the inverse association between cancer risk and intake of fruits and vegetables. These health benefits have been linked to the additive and synergistic combination of phytochemicals in fruits and vegetables. Cranberries have been shown to possess anti-carcinogenic activities such as inhibition of growth of several cancer cell lines, and inhibition of ornithine decarboxylase (ODC) activity in vitro. However, the molecular mechanisms of the anti-cancer properties of cranberry phytochemical extracts have not been completely understood. Our data showed that cranberry phytochemical extracts significantly inhibited human breast cancer MCF-7 cell proliferation at doses of 5 to 30mg/mL (P<0.05). Apoptotic induction in MCF-7 cells was observed in a dose-dependent manner after exposure to cranberry phytochemical extracts for 4h. Cranberry phytochemical extracts at a dose of 50mg/mL resulted in a 25% higher ratio of apoptotic cells to total cells as compared to the control groups (P<0.05). Cranberry phytochemical extracts at doses from 10 to 50mg/mL significantly arrested MCF-7 cells at G0/G1 phase (P<0.05). A constant increasing pattern of the G1/S index was observed in the cranberry extract treatment group while the G1/S ratio of the control group decreased concomitantly between 10 and 24h treatment. After 24-h exposure to cranberry extracts, the G1/S index of MCF-7 cells was approximately 6 times higher than that of the control group (P<0.05). These results suggest that cranberry phytochemical extracts possess the ability to suppress the proliferation of human breast cancer MCF-7 cells and this suppression is at least partly attributed to both the initiation of apoptosis and the G1 phase arrest.     

Ganoderma lucidum extract induces cell cycle arrest and apoptosis in MCF-7 human breast cancer cell

Although the pharmacology and clinical application of water extracts of Ganoderma lucidum have been extensively documented, little is known regarding its alcohol extract. In the present study, the anti-tumor effect of an alcohol extract of Ganoderma lucidum was investigated using MCF-7 cells. We found that the alcohol extract of Ganoderma lucidum inhibited cell proliferation in a dose- and time-dependent manner, which might be mediated through up-regulation of p21/Waf1 and down-regulation of cyclin D1. Furthermore, this compound can directly induce apoptosis in MCF-7 cells, which might be mediated through up-regulation of a pro-apoptotic Bax protein and not by the immune system. Our findings suggest that there are multiple mechanisms underlying the anti-tumor effects of Ganoderma lucidum.     

三氧化二砷对人肺癌细胞株增殖和凋亡的影响

目的 探讨三氧化二砷（Ａｓ２Ｏ３）对人肺癌的潜在性治疗作用及可能的机制。方法 选用人肺癌细胞株ＧＬＣ－８３,运用细胞培养法、ＭＴＴ法、流式细胞术（ＦＣＭ）检测细胞生长曲线、细胞增殖、细胞周期和细胞凋亡。结果 三氧化二砷可明显抑制ＧＬＣ－８２细胞的增殖,其抑制作用具有时－效和量－效关系。当４．０μｍｏｌ／Ｌ三氧化二砷处理ＧＬＣ－８２细胞９６小时,增殖抑制率达８１．０５％,ＦＣＭ检测肿瘤细胞ＤＮＡ含量,观察到三氧化二砷ＧＬＣ－８２细胞周期阻滞于Ｇ２／Ｍ期,细胞周期进程变慢,同时出现剂量依赖性Ｇ１峰。结论 三氧化二砷能有效地抑制人肺癌细胞株ＧＬＣ－８２的增殖,其可能的机制与三氧化二砷使细胞阻滞于Ｇ２／Ｍ期并诱导其调亡有关。     

替尼泊甙和羟基喜树碱对胃癌BGC-823细胞增殖和凋亡的作用

背景与目的:研究显示,拓扑异构酶(topoisomerase,Topo)Ⅰ和Ⅱ抑制剂联合应用有一定的协同抗肿瘤作用,但两物联合应用治疗胃癌的报道较少.本研究探讨Topo Ⅰ抑制剂羟基喜树碱(hydroxycamptothecin,HCPT)和TopoⅡ抑制剂替尼泊甙(teniposide,VM-26)联合应用对胃癌细胞BGC-823的体外抑制作用和诱导细胞凋亡的作用.方法:利用噻唑蓝(MTT)法测定HCPT、VM-26单药及联合用药对BGC-823细胞的体外抑制作用,流式细胞仪测定细胞凋亡.结果:1.963～31.413 μmol/L VM-26对BGC-823细胞的抑制率为15.99%～80.83%;1.963 μmol/LVM-26处理细胞0、12、24、48 h,细胞凋亡率分别为3.90%、4.42%、7.60%、17.70%.8.577～137.227 μmol/L HCPT对细胞的抑制率为7.89%～70.32%,8.577 μmol/LHCPT处理0、12、24、48 h后,细胞凋亡率分别为2.80%、8.50%、10.50%、13.30%.联合用药时抑制率为21.32%～87.74%,凋亡率为2.80%、15.50%、15.70%、20.20%.联合用药指数为1.293.结论:HCPT与VM-26单药可抑制BGC-823细胞增殖,诱导细胞凋亡;HCPT与VM-26联合应用在胃癌细胞中产生拮抗作用.     

唑来膦酸对肺癌95D细胞周期阻滞和诱导凋亡的作用机制

目的 研究唑来膦酸对肺癌95D细胞的细胞周期阻滞及诱导凋亡作用.方法 采用二苯基溴化四氮唑蓝法检测唑来膦酸对肺癌95D细胞增殖的影响;采用流式细胞仪检测唑来瞵酸作用后肺癌95D细胞的细胞周期和凋亡的变化,并在光镜和电镜下观察细胞凋亡的形态变化;采用Western blot和逆转录聚合酶链反应检测唑来膦酸对肺癌95D细胞凋亡相关基因ERK1/ERK2、bcl-2、bax和survivin表达水平的影响.结果 唑来膦酸对肺癌95D细胞的增殖抑制作用呈时间和浓度依赖性,94.0% μmol/L唑来膦酸作用6 d时,抑制率达89.8%.随着唑来膦酸作用时间的延长,G1期细胞数量增多,G2期和S期细胞减少,凋亡细胞的数最逐渐增多.光镜和电镜均观察到细胞出现了相应的凋亡形态学变化.细胞凋亡相关基因ERK、bcl-2和survivin的表达水平下降,bax的表达水平增高.结论 唑来膦酸对肺癌95D细胞的细胞周期有阻滞作用,并能诱导其凋亡.     

奥沙利铂联合替尼泊甙对胃癌BGC-823细胞的生长抑制和诱导凋亡的协同作用(英文)

Objective:The aim of this study was to investigate the synergistic effects of oxaliplatin and teniposide on proliferation and apoptosis of gastric cancer cell line BGC-823.Methods:MTT assay was carried to examine the inhibition rate of oxaliplatin and teniposide on gastric cancer cell line BGC-823 with various concentrations separately and associatively.The apoptosis rate of BGC-823 cells under the treatment of oxaliplatin or/and teniposide was examined by flow cytometry.The expression level of livin, an ap...     

PPARγ活化诱导细胞凋亡和周期阻滞来抑制结肠癌细胞生长

目的探讨过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptorγ,PPARγ)在结肠癌细胞株HT-29中的表达及其活化后对结肠癌细胞生长的影响。方法通过RT-PCR和Western blot检测HT-29中PPARγmRNA及蛋白质的表达,四甲基偶氮唑盐(MTT)微量酶反应比色法检测PPARγ配体罗格列酮(rosiglitazone,Rosi)和15d-PGJ2对HT-29细胞生长的影响,荧光显微镜、TUNEL法观察Rosi和15d-PGJ2激活PPARγ后诱导HT-29细胞凋亡形态和生化改变,流式细胞仪(FCM)以碘化丙啶(PI)单染法检测细胞周期。结果RT-PCR及Western blot检测结果表明结肠癌细胞HT-29中存在PPARγmRNA及蛋白质的表达。MTT结果显示Rosi和15d-PGJ2可抑制HT-29细胞生长,且具有时间、剂量依赖效应。荧光显微镜、TUNEL检测显示PPARγ活化后HT-29细胞出现典型的凋亡现象,10μmol/L Rosi或15d-PGJ2作用48h后的细胞凋亡率分别为(17.3±1.9)%及(20.8±2.9)%,与对照组(3.86±0.49)%相比差异均具有统计学意义(P<0.05)。FCM结果显示Rosi和15d-PGJ2激活PPARγ后诱导细胞周期阻滞于G0/G1期,与对照组相比差异有统计学意义(P<0.05)。结论结肠癌细胞HT-29表达PPARγ,其活化可通过诱导细胞凋亡及细胞周期阻滞,抑制结肠癌细胞增长。因此,PPARγ可能为结肠癌治疗的一个新靶点。     

Exogenous morphine inhibits human gastric cancer MGC- 803 cell growth by cell cycle arrest and apoptosis induction

Morphine is not only an analgesic treating pain for patients with cancer but also a potential anticancer drug inhibiting tumor growth and proliferation. To gain better insight into the involvement of morphine in the biological characteristics of gastric cancer, we investigated effects on progression of gastric carcinoma cells and the expression of some apoptosis-related genes including caspase-9, caspase-3, survivin and NF-κB using the MGC-803 human gastric cancer cell line. The viability of cells was assessed by MTT assay, proliferation by colony formation assay, cell cycle progression and apoptosis by flow cytometry and ultrastructural alteration by transmission electron microscopy. The influences of morphine on caspase-9, caspase-3, survivin and NF-κB were evaluated by semi-quantitative RT-PCR and Western blot. Our data showed that morphine could significantly inhibit cell growth and proliferation and cause cell cycle arrest in the G2/M phase. MGC-803 cells which were incubated with morphine also had a higher apoptotic rate than control cells. Morphine also led to morphological changes of gastric cancer cells. The mechanism of morphine inhibiting gastric cancer progression in vitro might be associated with activation of caspase-9 and caspase-3 and inhibition of survivin and NF-κB.     

LINC01060对人胃癌BGC-823细胞系生物学行为的作用研究

目的:探索LINC01060对人胃癌BGC-823细胞系生物学行为的作用。方法:生信数据分析LINC01060在胃癌中的表达情况,建立LINC01060过表达和沉默的稳转BGC-823细胞株,Real time PCR验证LINC01060的表达,CCK-8检测细胞增殖,流式细胞术检测细胞凋亡,划痕实验检测细胞迁移,Transwell检测细胞侵袭,Western blot检测EMT相关蛋白E-cadherin、Vimentin、N-cadherin和Snail蛋白的表达。结果:GEO数据库发现,LINC01060在胃癌中低表达（P＜0.05）;Real time PCR结果显示,沉默组LINC01060的表达显著下调（P＜0.05）,过表达组LINC01060的表达显著上调（P＜0.05）;CCK-8显示,过表达组细胞增殖能力显著下降（P＜0.05）;流式细胞术显示,过表达组细胞凋亡比例显著上升（P＜0.05）;Transwell结果显示,过表达组细胞侵袭能力显著下降（P＜0.05）;细胞划痕显示,过表达组细胞迁移能力显著下降（P＜0.05）;Western blot显示,过表达组E-cadherin蛋白表达水平显著上调（P＜0.05）,Vimentin、N-cadherin、Snail蛋白表达水平显著下调（P＜0.05）;以上实验沉默组均相反。结论:LINC01060在人胃癌BGC-823细胞中发挥促进凋亡,抑制增殖、侵袭、迁移和EMT的作用。