三氧化二砷诱导肝癌细胞(BEL-7402)凋亡研究

目的探寻三氧化二砷对人肝癌细胞株（BEL-7402）的增殖抑制作用及诱导其凋亡的作用效果.方法采用噻唑蓝（MTT）法判定As2O3对BEL-7402细胞的细胞毒作用;用AO/EB荧光染色观察细胞的形态学变化;采用单细胞电泳和流式细胞光度术研究细胞凋亡和细胞周期的变化.结果三氧化二砷能显著抑制BEL-7402细胞的生长,10,20μg/ml可出现较为典型的细胞凋亡的形态变化,细胞发生彗星样电泳图像,并且出现典型的亚二倍体凋亡峰,其作用表现为细胞周期特异性,主要作用于细胞周期的S期.结论三氧化二砷对肝癌BEL-7402细胞具有很强的细胞毒作用,诱导细胞凋亡是其主要作用机制之一.     

去蛋氨酸环境中多种化疗药对肝癌细胞株的杀伤作用

目的 探索肝癌细胞和肝细胞在缺乏蛋氨酸环境中的生长状况和去蛋氨酸状况与多种化疗药物联合应用是否可增加对肝癌细胞的杀伤作用。方法 将肝癌细胞株BEL-7402、肝细胞株L-02分别置于不含蛋氨酸含同型半胱氨酸培养基(Met^-Hcy^ )和含蛋氨酸不含同型半胱氨酸培养基(Met^ Hcy^-)中培养，MTT法检测两组细胞增值状态及在不同培养基中分别加入ADM、CDDP、5-FU、MMC和MTX化疗药物后，对各组细胞生长的抑制程度。结果 L-02在Met^-Hcy^ 和Met^ Hcy^-中生长无差异；Met^-Hcy^ 未增加5-FU、ADM、CDDP、MMC和MTX对L-02的毒性作用。与Met^ Hcy^-组相比，在Met^-Hcy^ 培养基中BEL-7402细胞生长受抑。Met^-Hcy^ 与5-FU相结合可显著提高对BEL-7402细胞的杀伤作用，与ADM、CDDP、MMC和MTX结合未提高对BEL-7402细胞的杀伤作用。结论 肝癌细胞BEL-7402细胞具有Met依赖性。Met^-Hey^ 与周期特异性化疗药物5-FU相结合可显著提高其对肝癌细胞BEL-7402的杀伤作用。     

苦参碱对肝癌细胞增殖抑制作用研究

目的在体外观察苦参碱对BEL-7402肝癌细胞增殖的影响。方法用含不同浓度（0～3.2mmol/L）苦参碱的培养液在体外培养BEL-7402细胞24h,通过MTT试验测定其对BEL-7402细胞增殖的影响作用。结果在上述使用浓度范围内,苦参碱对BEL-7402细胞增殖的抑制率为0～36.7%。结论苦参碱在体外可抑制BEL-7402细胞的增殖,且呈剂量依赖性。     

103Pd粒子联合加热41℃对人肝癌细胞7402细胞毒作用的实验研究

目的 研究^103Pd粒子与加热41℃对人肝癌细胞株(Bel-7402)的细胞毒作用，了解^103Pd粒子对热疗后残存的肝癌细胞的细胞毒作用。方法 以体外培养的人肝癌细胞-7402为研究对象，采用对照分组、水浴加温法、体外细胞毒试验(MTT)、凋亡实验(Hoechst33258荧光染色法)的方法以及方差分析，观察^103Pd粒子放疗联合加热41℃对Bel-7402细胞的生长抑制作用。结果 ^103Pd粒子对41℃加热后的人肝癌细胞-7402有明显的细胞抑制作用，强于单纯^103Pd粒子放疗。结论 人肝癌细胞-7402对^103Pd粒子放疗敏感，^103Pd粒子联合加热41℃有增效作用。提示：^103Pd粒子对热疗后残存的肝癌细胞有细胞毒作用。     

阿魏蘑菇提取物抗肿瘤作用的实验研究

目的系统测试阿魏蘑菇提取物抗肿瘤的效应,探讨其抗癌机制。方法应用MTT法检测提取物对泌尿生殖系统中4个肿瘤细胞株生长的抑制作用;分别检测提取物对S180小鼠实体瘤生长的抑制率和S180小鼠腹水瘤小鼠的生命延长率;以肿瘤细胞凋亡为切入点,研究提取物抗肿瘤机制。结果提取物高浓度具有明显抑制肿瘤细胞生长作用(P<0.01),极低浓度未见抑制肿瘤细胞生长作用。阿魏蘑菇提取物高中低剂量均有抑制S180实体瘤小鼠肿瘤生长的作用,均可明显延长S180腹水瘤小鼠的生存时间,且其延长小鼠生存时间呈剂量依赖关系。形态学观察发现提取物诱导了4种肿瘤细胞的凋亡;提取物诱导4种肿瘤细胞早期凋亡率明显高于未处理肿瘤细胞;提取物处理T24细胞后,baxc、aspase-3蛋白表达明显增强,bcl-2表达降低。结论阿魏蘑菇提取物有抗肿瘤作用,其机制可能与下调bcl-2表达和上调bax的表达,从而高表达caspase-3诱导肿瘤细胞凋亡有关。     

三氧化二砷对人肝癌多药耐药细胞Bel-7402作用相关机制的研究

目的研究As2O3诱导多药耐药人肝癌细胞Bel-7402/VCR凋亡及相关基因表达的情况。方法MTT法观察细胞生长和对VCR敏感性,应用形态学手段和流式细胞术检测细胞凋亡,应用免疫组织化学研究多药耐药相关基因表达的影响。结果As2O3对Bel-7402/VCR及Bel-7402细胞的生长抑制均呈明显的量效关系;Bel-7402/VCR及Bel-7402凋亡细胞百分比随着As2O3作用时间延长和作用浓度的增加明显增加;As2O3对Bel-7402/VCR及Bel-7402MDR1阳性表达率为81.6%和80.2%。结论As2O3在体外表现出较强的逆转肝癌多药耐药性作用。     

三氧化二砷对人肝癌多药耐药细胞Bel-7402作用相关机制的研究

目的研究As2O3诱导多药耐药人肝癌细胞Bel-7402／VCR凋亡及相关基因表达的情况。方法MTT法观察细胞生长和对VCR敏感性,应用形态学手段和流式细胞术检测细胞凋亡,应用免疫组织化学研究多药耐药相关基因表达的影响。结果As2O3对Bel-7402／VCR及Bel-7402细胞的生长抑制均呈明显的量效关系;Bel-7402／VCR及Bel-7402凋亡细胞百分比随着As2O3作用时间延长和作用浓度的增加明显增加;As2O3对Bel-7402／VCR及Bel-7402MDR1阳性表达率为81．6％和80．2％。结论As2O3在体外表现出较强的逆转肝癌多药耐药性作用。     

白毛藤对人肝癌Bel7402细胞增殖抑制及凋亡作用

目的 探讨白毛藤体外对人肝癌Bel7402细胞增殖与凋亡的影响及其作用机制.方法 采用四甲基偶氮噻唑蓝(MTT)法和荧光显微镜观察白毛藤对人肝癌Bel7402细胞的增殖抑制作用和其诱导Bel7402细胞凋亡情况;用反转录-聚合酶链式反应(RT-PCR)检测survivin、bax基因表达量的变化.结果 随白毛藤浓度增加、作用时间延长,对人肝癌Bel7402细胞的增殖抑制作用均明显增强(P＜0.01);4,8,16 mg/mL白毛藤组人肝癌Bel7402细胞凋亡率分别为10.98%、17.02%、26.78%,均高于对照组6.18%(P＜0.01);作用48 h后,4,8,16 mg/mL白毛藤组人肝癌Bel7402细胞中survivin基因灰度比值分别为(0.43±0.01)、(0.38±0.02)、(0.29±0.01).均低于对照组(0.65±0.05),差异均有统计学意义(P＜0.01);4,8,16 mg/mL,白毛藤组人肝癌Bel7402细胞中bax基因灰度比值分别为(0.47±0.05)、(0.56±0.04)、(0.73±0.06),均高于对照组(0.36±0.02),差异均有统计学意义(P＜0.01).结论 白毛藤可抑制人肝癌Bel7402细胞增殖、诱导其细胞凋亡,并能调控survivin及bax基因的表达.     

野蔷薇果提取物对不同肿瘤细胞抑制作用

目的探讨野蔷薇果提取物抗肿瘤作用。方法应用四甲基偶氮噻唑蓝(MTT)法,检测提取物对4株肿瘤细胞株(hela、bcap、T24、ho8910)生长抑制作用;通过体内试验分别检测提取物对S180小鼠实体瘤肿瘤生长的抑制率和腹水瘤小鼠生命延长率,观察了提取物对肿瘤细胞凋亡影响。结果野蔷薇果提取物具有抑制S180实体瘤小鼠肿瘤生长的作用,高、中、低3个剂量组肿瘤生长抑制率分别为46.28%,40.45%和65.7%,腹水瘤小鼠生命分别延长44.7%,48.2%和8.2%;提取物诱导4种肿瘤细胞凋亡率在16.7%～72.0%,明显高于未处理细胞(P0.05),bax、caspase-3蛋白表达明显增强,bcl-2表达降低。结论野蔷薇果提取物对肿瘤细胞生长具有抑制作用,其作用机制可能与下调bcl-2、上调bax以及高表达caspase-3诱导肿瘤细胞凋亡有关。     

米氏凯伦藻甲醇氯仿提取物的细胞毒性分析

目的研究米氏凯伦藻提取物对3株肿瘤细胞的增殖抑制作用,分析米氏凯伦藻对人类健康的可能威胁。方法以体外培养的人宫颈癌细胞HeLa、人肝癌细胞HepG2和人肺癌细胞A549为研究对象,采用MTT比色法检测米氏凯伦藻中提取物对3株细胞增殖的影响;观察毒素在不同时间(12、24、36和48h)下对肿瘤细胞增殖的抑制情况;采用免疫荧光染色法分析比较3株细胞膜上GM1的相对含量。结果米氏凯伦藻提取物对3种肿瘤细胞的增殖均具有显著的抑制作用。与肝癌细胞HepG2相比,宫颈癌细胞HeLa和肺癌细胞A549对其更为敏感(P<0.01)。肝癌细胞HepG2、宫颈癌细胞HeLa和肺癌细胞A549细胞膜上GM1的相对含量差异不大(P>0.05)。结论米氏凯伦藻提取物具有明显的细胞毒性,对人类健康存在潜在威胁。米氏凯伦藻提取物在细胞膜上作用靶点比较复杂。     

新疆天然植物黑加仑对食管癌细胞增殖、凋亡的影响

目的:观察新疆天然植物黑加仑水提物对食管癌细胞的影响及其作用机制。方法:采用四甲基偶氮唑蓝(MTT)法测定不同浓度(10-1、10-2、10-3g/ml)黑加仑水提物作用不同时间(12、24、36h)对人食管癌细胞株Eca109的细胞存活量的影响,以大鼠正常肝细胞(BRL)为正常对照细胞株;用Bradford法测定肿瘤细胞蛋白质合成的变化;采用流式细胞技术(FCM)观察黑加仑作用后肿瘤细胞周期变化及凋亡情况;通过倒置显微镜观察细胞的形态变化。结果:黑加仑水提物10-1g/ml组对人食管癌细胞株Eca109作用24h后,癌细胞的生长和蛋白合成均有明显抑制作用(P<0.05),对正常肝细胞的存活量及蛋白合成均无影响;流式细胞仪检测二倍体峰前出现凋亡峰,细胞凋亡率为49.6%;并且癌细胞呈现典型的凋亡细胞形态。结论:黑加仑水提物可通过诱导细胞凋亡而抑制人食管癌细胞株Eca109细胞的生长。     

黑米皮提取物对PC-3细胞增殖的抑制作用

目的观察黑米皮提取物对人前列腺癌PC-3细胞增殖的抑制作用并探讨其机制。方法用不同浓度的黑米皮提取物(20,150和300μg/ml)处理PC-3细胞,通过MTT法、流式细胞仪、western blot法研究提取物对细胞的影响。结果处理组的细胞增殖率随着提取物作用浓度和时间的增加而降低(P<0.05);与对照组相比,处理组细胞凋亡率增加,差异有统计学意义(P<0.05);细胞周期中,G1期细胞百分比显著增多,S、G2-M期细胞显著减少,细胞被阻滞在G1-S期。与对照组相比,处理组细胞的p-p38、p-JNK的蛋白表达则随着提取物浓度的增加而增强。结论黑米皮提取物能够抑制人前列腺癌PC-3细胞的增殖,改变细胞周期的分布,激活JNK、p38通路,诱导细胞凋亡。     

Effects of germinated brown rice extracts with enhanced levels of GABA on cancer cell proliferation and apoptosis

In the present work we investigated the effects of brown rice extracts on proliferation and apoptosis of cancer cells. Brown rice extracts were prepared using nongerminated brown rice versus germinated brown rices. Mouse leukemia L1210 cells, human acute lymphoblastic leukemia Molt4 cells, and human cervical cancer HeLa cells were treated with either nongerminated brown rice extract (N ex), water-germinated extract (W ex), chitosan-germinated extract (C ex), glutamic acid-germinated brown rice extract (G ex), or chitosan/glutamic acid-germinated brown rice extract (CG ex). The concentrations of gamma-aminobutyric acid (GABA) in the G ex and CG ex were three and 3.3 times higher than the GABA concentration in the N ex, respectively. The G ex and CG ex retarded significantly the proliferation rates of L1210 and Molt4 cells, and the highest retardation rate was with CG ex. In addition, the G ex and CG ex enhanced significantly apoptosis of the cultured L1210 cells, but no significant apoptosis was seen with the other extracts, which have lower concentrations of GABA than G ex and CG ex. These results show that brown rice extracts with enhanced levels of GABA have an inhibitory action on leukemia cell proliferation and have a stimulatory action on the cancer cell apoptosis.     

吲哚-3-甲醇抑制人肝癌细胞株SMMC-7721生长作用及机理探讨

目的探讨吲哚-3-甲醇(I3C)抑制人肝癌细胞株SMMC-7721生长作用及机理。方法采用人肝癌细胞株SMMC-7721制备荷瘤裸鼠;通过测量肿瘤大小和病理学检测,观察I3C对肿瘤细胞的抑制作用;采用流式细胞仪检测肿瘤细胞周期、线粒体膜电位;采用Western blot法检测相关蛋白的表达。结果经I3C作用后,SMMC-7721细胞生长变慢,细胞周期、线粒体膜电位、相关蛋白的表达改变。结论 I3C能够抑制SMMC-7721细胞生长,细胞周期和细胞凋亡相关信号分子可能在其中发挥了一定作用。     

A heat-stable extract from Mucuna stimulates the differentiation of bone marrow cells into dendritic cells and induces apoptosis in cancer cells

Pine cone extract is known to induce differentiation of human mononuclear cells into dendritic cells (DCs) and also to induce apoptosis in human cancer cells. In the present study, we screened edible plants that contain components with biological activities similar to or more potent than those of pine cone extract. We found that Mucuna (Mucuna pruviens var. utilis) contains a DC differentiation/maturation-inducing activity and a component that induces apoptosis in human cancer cell lines. Mucuna extract specifically stimulated differentiation of BM cells to immature DCs. Marked production of IL-6 was observed by sequential treatment with at least 10 μg/mL of Mucuna extract followed by LPS. The sequential treatment with Mucuna extract followed by LPS produced a much higher ratio of IL-12 to IL-6 and a lower ratio of TNF-α to IL-6 than that obtained by sequential treatment with a medicinal mushroom Phellinus linteus extract and then LPS. The DC differentiation/maturation activity and the component inducing apoptosis in cancer cells were separable by column chromatography.     

天然红心蛋中类胡萝卜素提纯物对人肝癌细胞增殖和凋亡的影响

目的:探讨天然红心蛋中类胡萝卜素提纯物(carotenoidsextractsfromnaturalredyolk,CENRY)对人肝癌细胞QGY-7703细胞增殖和凋亡的影响。方法:采用体外培养,细胞增殖实验,激光扫描共聚焦显微镜(LSCM)观察细胞形态及钙离子浓度变化,采用流式细胞术检测细胞周期等。结果:CENRY1.0、0.5、0.25μmol/L三个剂量能显著抑制人肝癌QGY-7703细胞增殖,细胞凋亡指数随剂量增加及时间延长而上升;CENRY对QGY-7703细胞存在G2/M期阻滞作用;处理组细胞内Ca2+浓度极显著地升高。结论:CENRY能抑制QGY-7703增殖、诱导凋亡。     

转染人内皮抑素对卵巢癌细胞A2780体内外生长的影响

目的:探讨人内皮抑素(hum an endostatin,hES)对人卵巢癌细胞株A2780体内、外生长的影响及其可能机制。方法:高保真PCR从含有人内皮抑素基因的人肝脏组织中扩增出人内皮抑素编码区克隆入pcDNA3.1载体中,构建pcDNA3.1-ES重组质粒;将此重组质粒转染人卵巢癌细胞A2780中,观察A2780细胞生长。建立人卵巢癌裸鼠模型,随机将其分为两组:hES转染细胞组(A组,10只),对照组(B组,10只)。1月后,测量瘤体积、计算抑瘤率;标本行HE染色分析组织学形态;同时用W estern b lot检测hES、凋亡抑制蛋白bc l-2的表达;流式细胞仪检测转染对裸鼠移植瘤细胞凋亡率的影响。结果:hES转染A2780细胞后,细胞生长速率减慢。转染hES的裸鼠肿瘤生长较对照组明显减慢(P<0.001),抑瘤率为74%;HE染色及W estern b lot提示:转染后(A组)的组织学形态与转染前(B组)有显著差异;A组hES蛋白水平明显高于B组,而bc l-2蛋白水平明显低于B组;流式结果显示转染使肿瘤细胞的凋亡显著增加(P<0.05)。结论:转染hES体内、外均可抑制卵巢癌A2780细胞的生长,其机制与其促进了凋亡有关。     

Loquat (Eriobotrya japonica) extracts suppress the adhesion, migration and invasion of human breast cancer cell line

We examined the inhibitory effects of loquat methanol extract on the adhesion, migration, invasion and matrix metalloproteinase (MMP) activities of MDA-MB-231 human breast cancer cell line. Cells were cultured with DMSO or with 10, 25, or 50 µg/ml of loquat methanol extract. Both leaf and seed extracts significantly inhibited growth of MDA-MB-231 cells in a dose-dependent manner, although leaf extract was more effective. Adhesion and migration were significantly inhibited by loquat extracts in a dose-dependent manner. Loquat extract also inhibited the invasion of breast cancer cells in a dose-dependent manner and leaf extract was more effective than seed extract. MMP-2 and MMP-9 activities were also inhibited by loquat extract. Our results indicate that methanol extracts of loquat inhibit the adhesion, migration and invasion of human breast cancer cells partially through the inhibition of MMP activity and leaf extract has more anti-metastatic effects in cell based assay than seed extract. Clinical application of loquat extract as a potent chemopreventive agent may be helpful in limiting breast cancer invasion and metastasis.     

Anthocyanin-rich extract from Aronia meloncarpa E induces a cell cycle block in colon cancer but not normal colonic cells

Anthocyanin-rich extracts, potent antioxidants and commercially available food coloring agents, have been reported to inhibit growth of various cancer cell lines. We investigated the effect of semipurified anthocyanin-rich extract from fruits of Aronia meloncarpa, on normal colon and colon cancer cell lines. A 24-h exposure to 50 mg monomeric anthocyanin/ml of Aronia extract resulted in 60% growth inhibition of human HT-29 colon cancer cells. The treated cells showed a blockage at G1/G0 and G2/M phases of the cell cycle. The cell cycle arrest coincided with an increased expression of the p21WAF1 and p27KIP1 genes and decreased expression of cyclin A and B genes. Prolonged exposure to the extract resulted in no further change in the cell number, indicating a cytostatic inhibition of cell growth. NCM460 normal colon cells demonstrated <10% growth inhibition at the highest concentration of 50 mg/ml extract. A 35% decrease in the cyclooxygenase-2 gene expression was observed within 24 h of exposure of HT-29 cells but did not translate into decreased protein levels or protein activity. These results support the need for further research to identify the specific component(s) in this extract that suppress cancer cell growth and the genes affected by these natural compounds.