白杨素增强rhsTRAIL对人胃癌SGC-7901细胞毒性

目的:研究白杨素(ChR)是否具有增强rhsTRAIL对人胃癌SGC-7901细胞毒性作用。方法:体外培养SGC-7901细胞。MTT比色法测定细胞毒性。碘化丙啶(PI)染色流式细胞术(FCM)分析细胞死亡率。结果:ChR、rhsTRAIL以及两者合用对SGC-7901细胞活性的半数抑制浓度(IC50值)分别为134μmol/L、402ng/mL和47ng/mL,合并用药效应的CI值是0.4676。ChR40μmol/L、rhsTRAIL100ng/mL以及两者合用的细胞死亡率分别为4.65%±0.58%、3.60%±0.16%和49.87%±4.27%。结论:亚细胞毒性浓度的白杨素具有增强rhsTRAIL对人胃癌SGC-7901细胞毒性作用。     

白杨素对5- 氟尿嘧啶诱导人肝癌Bel-7402 细胞凋亡的增敏机制研究

目的 探讨白杨素对5- 氟尿嘧啶（5-FU）诱导肝癌Bel-7402 细胞凋亡的增敏作用。方法 白 杨素单独或联合5-FU 作用Bel-7402 细胞,用MTT 法检测细胞活力;流式细胞术检测细胞凋亡与周期分 布变化;Western blotting 检测Bcl-2、Bax、Bad 及Cleaved Caspase-3 蛋白。结果 50 ～ 250μmol/L 白杨 素作用Bel-7402 细胞48 h 呈浓度依赖性的抑制细胞活力,且100μmol/L 白杨素组细胞存活率为（82.261± 7.793）%。100μmol/L 白杨素单独作用24 h 对Bel-7402 细胞活力和凋亡均无影响（P >0.05）。与空白对照 组比较,100μmol/L 白杨素作用24 h 后,不同浓度5-FU（12.5、25.0 及50.0μg/ml）作用48 h 对Bel-7402 细胞活力的抑制作用增强（P <0.05）;25.0μg/ml 5-FU 诱导Bel-7402 细胞凋亡率增加（P <0.05）;G2/M 期 细胞比例升高（P <0.05）;促凋亡蛋白Bax、Bad 及Cleaved Caspase-3 蛋白表达水平上调（P <0.05）;抗凋亡 蛋白Bcl-2 表达水平下调（P <0.05）。结论 白杨素增强5-FU 诱导人肝癌Bel-7402 细胞凋亡,其机制可能 与线粒体凋亡通路激活有关。     

白杨素敏化TRAIL诱导人胃癌SGC-7901细胞凋亡

目的:研究白杨素(ChR)是否具有增敏肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导人胃癌SGC-7901细胞凋亡作用.方法:体外培养SGC-7901细胞.碘化丙啶(PI)染色流式细胞术(FCM)分析细胞凋亡率,琼脂糖凝胶电泳检测DNA梯形条带.结果:ChR(40 μmol/L)、TRAIL(100 ng/mL)以及两...     

芹菜素增强TRAIL诱导人宫颈癌HeLa细胞凋亡

目的:研究芹菜素(API)与重组人可溶性肿瘤坏死因子相关凋亡诱导配体(TRAIL)合用对人宫颈癌HeLa细胞凋亡的影响。方法:体外培养人宫颈癌HeLa细胞。碘化丙啶(PI)染色流式细胞术(FCM)定量分析sub-G1百分率。DNA琼脂糖凝胶电泳观察细胞DNA梯形条带。结果:API(20μmol/L)和TRAIL(20ng/mL)以及两者合用作用48h的人宫颈癌HeLa细胞sub-G1百分率分别是3.56%±0.20%、6.69%±0.40%和59.8%±4.20%;DNA琼脂糖凝胶电泳显示:20μmol/LAPI与20ng/mL的TRAIL联合处理,展示出典型DNA梯形条带图谱。结论:芹菜素具有增强TRAIL诱导HeLa细胞凋亡作用。     

Bim介导白杨素诱导肝癌Hep 3B细胞凋亡

目的:研究白杨素诱导人肝癌Hep 3B细胞凋亡是否涉及上调促凋亡蛋白Bim.方法:体外培养Hep3B细胞.碘化丙啶(PI)染色流式细胞术(FCM)分析细胞凋亡率.细胞凋亡ELISA试剂盒检测细胞组蛋白/DNA碎片.Western blot和小干扰RNA(siRNA)转染用于探索其作用机制.结果:白杨素显著诱导Hep3B凋亡,并上调Bim表达.Bim特异性siRNA能有效拮抗白杨素诱导Hep 3B细胞凋亡作用.结论:白杨素具有诱导人肝癌Hep3B细胞凋亡作用,其作用机制涉及上调Bim表达.     

芹菜素敏化TRAIL诱导人卵巢癌CoC1细胞凋亡

目的:研究芹菜素(API)是否具有敏化肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导人卵巢癌CoC1细胞凋亡作用。方法:体外培养CoC1细胞。碘化丙啶(PI)染色流式细胞术(FCM)定量分析sub-G1细胞百分率。ELISA法测定细胞Caspase-3活性。结果:API(20μmol/L)和TRAIL(20ng/mL)以及两者合用48h的sub-G1细胞百分率分别是8.83%±2.33%、8.32%±2.80%和69.50%±4.65%;人卵巢癌CoC1细胞Caspase-3的活性分别是培养基组的1.3、1.4和6.5倍。结论:亚细胞毒性浓度的芹菜素具有增强TRAIL诱导人卵巢癌CoC1细胞凋亡作用。     

ERK1/2介导的bFGF对肝癌细胞系Bel-7402细胞增殖和凋亡的影响

目的:观察Ras-Raf-ERK1/2途径介导的碱性成纤维细胞生长因子(bFGF)对肝癌细胞系Bel-7402细胞增殖和凋亡的影响,探讨bFGF及其信号转导途径与肝癌发生发展的关系。方法:以bFGF和PD98059处理Bel-7402细胞,流式细胞术检测细胞周期与凋亡情况;MTT法检测细胞增殖情况;Western blot检测细胞外信号调节蛋白激酶(ERK1/2)的活化。结果:bFGF处理,诱导细胞进入S期[S期细胞比例(27.49±0.72)%→(42.45±1.06)%];细胞增殖比明显增加,且与bFGF水平成剂量依赖关系,bFGF浓度为25 ng/ml时细胞增殖比最高为129%;使无血清饥饿诱导的凋亡细胞比例下降[(28.89±3.13)%(→1.70±2.10)%];时、量效依赖性地诱导ERK1/2活性增高。ERK激活性蛋白激酶(MEK1)抑制剂PD98059可抑制bFGF的这些作用。结论:bFGF通过Ras-Raf-ERK1/2途径介导,加速肝癌Bel-7402细胞的细胞周期进程,促进细胞增殖,抵抗无血清饥饿诱导的凋亡。在肝癌发生发展过程中,bFGF信号传递发挥了重要的作用。     

薯蓣皂苷对肝癌细胞Bel-7402和正常人肝细胞LO2增殖和凋亡的影响

目的研究薯蓣皂苷(dioscin)对肝癌细胞Bel-7402及正常人肝细胞LO2增殖和凋亡的影响及其可能机制。方法将0.5~16μmol/L薯蓣皂苷干预Bel-7402肝癌细胞和LO2肝细胞24 h,使用MTT法检测细胞增殖抑制作用,倒置显微镜观察细胞形态变化,Hoechst33258染色法观察细胞凋亡的形态学变化,JC-1染色法检测线粒体膜电位水平变化;Western blot蛋白质印迹法观察薯蓣皂苷对两种细胞中Bcl-2、Bax蛋白表达的影响。结果与阴性对照组比较,薯蓣皂苷各剂量组可显著抑制Bel-7402肝癌细胞和LO2肝细胞增殖,并呈剂量依赖性,其作用于Bel-7402肝癌细胞的IC50为2.04μmol/L,作用于LO2肝细胞的IC50为2.76μmol/L;与阴性对照组比较,1μmol/L、2μmol/L薯蓣皂苷组Bel-7402肝癌细胞及LO2肝细胞分布密度降低,细胞出现变圆脱落死亡,细胞边界模糊,并诱导细胞发生凋亡。JC-1染色结果显示,薯蓣皂苷作用于两种细胞后线粒体膜电位均显著降低(P<0.01)。Western blot蛋白质印迹法检测结果显示,薯蓣皂苷均可抑制两种细胞中Bcl-2蛋白的表达,上调Bax蛋白的表达(P<0.05,P<0.01)。结论薯蓣皂苷能够抑制Bel-7402肝癌细胞和LO2肝细胞的体外增殖,其机制可能是降低线粒体膜电位,抑制Bcl-2蛋白表达,上调Bax蛋白的表达,诱导细胞发生凋亡。提示薯蓣皂苷在发挥抗肝癌作用的同时亦对肝细胞具有损伤作用。     

人参皂苷Rh2对人肝癌Bel-7402细胞增殖和凋亡的影响

目的观察人参皂苷Rh2对人肝癌Bel-7402细胞增殖和凋亡的影响,并初步研究其机制。方法将Bel-7402细胞与人参皂苷Rh(2200、100和50μg/mL)共同培养48 h。MTT法检测人参皂苷Rh2对Bel-7402细胞增殖的影响;流式细胞仪分析细胞周期中各时期的细胞百分数;TUNEL技术检测其对Bel-7402细胞凋亡的影响;Western blotting法检测Bel-7402细胞中Bcl-2、Bax的蛋白表达。结果 MTT结果显示,人参皂苷Rh2能抑制Bel-7402细胞的生长;流式细胞检测显示,人参皂苷Rh2能使细胞停滞于G1期;TUNEL结果显示,人参皂苷Rh2能有效诱导Bel-7402细胞凋亡;Western blotting结果显示,人参皂苷Rh2能上调Bel-7402细胞中Bax的表达,下调Bcl-2的表达。上述结果均在一定程度上呈量效关系。结论人参皂苷Rh2能抑制Bel-7402细胞生长并促进其凋亡,其作用机制可能为上调Bax和下调Bcl-2。     

联合索拉非尼与TRAIL对人结肠癌HT-29细胞凋亡的影响

目的:研究多激酶抑制剂甲苯磺酸索拉非尼(SOR)是否具有增强肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导人结肠癌HT-29细胞凋亡作用。方法:体外培养HT-29细胞。碘化丙啶(PI)染色流式细胞术(FCM)定量分析亚二倍体(sub-G1)细胞百分率。比色法测定细胞Caspase-3活性。AO/EB双染色荧光显微镜观察凋亡细胞形态。结果:SOR(5μmol/L)和TRAIL(100ng/mL)以及两者合用作用48小时HT-29细胞的sub-G1百分率分别是4.65%±2.33%、5.29%±2.80%和42.6.50%±4.65%;HT-29细胞的Caspase-3的活性分别是培养基组的1.24、1.37和9.51倍。两者合用展示出细胞核染色质固缩和碎裂的典型凋亡细胞形态。结论:亚细胞毒性浓度的SOR具有增强TRAIL诱导人结肠癌HT-29细胞凋亡作用。     

硒蛋氨酸对肝癌细胞株SMMC7402凋亡的诱导作用

目的:研究硒蛋氨酸对肝癌细胞株SMMC7402生长的影响及其作用机制。方法:采用MTT比色法、流式细胞术、DNA片段分析法,研究硒蛋氨酸对肝癌SMMC7402细胞生长、增殖及凋亡的影响。结果:硒蛋氨酸呈时间-剂量依赖性方式抑制SMMC7402细胞生长增殖;G1期前出现亚倍体凋亡峰,细胞阻滞于G0/G1期;DNA电泳出现特征性凋亡梯带。结论:硒蛋氨酸可抑制肝癌细胞株SMMC7402的生长增殖,诱导细胞凋亡。     

丹皮酚对人肝癌Bel-7404的抑瘤效应及其机制

目的：探讨丹皮酚（Paeonol，Pae）抑制人肝癌细胞株Bel-7404增殖作用及其分子机制。方法：应用光学显微镜、MTT法检测经不同浓度、不同时间Poe处理Bel-7404细胞后的增殖抑制作用；DNA凝胶电泳法、TUNEL法检测细胞凋亡；用RT—PCR法、免疫细胞化学ABC法检测抑癌基因PTEN和致癌基因Akt表达水平。结果：与空白对照组比，Pae能抑制人肝癌细胞Bel-7404的增殖，诱导其凋亡，光镜下可见明显的凋亡细胞；DNA凝胶电泳显示出典型的凋亡特征；RT—PCR和免疫细胞化学检测显示，Poe处理Bel-7404后可使PTEN表达升高，Akt表达下降。结论：Pae具有抗Bel-7404细胞增殖的作用，其机制可能是通过抑制P13K通路，增强诱导其凋亡有关。     

Anti-tumor effect and mechanism of Paeonol on the hepatocellular carcinoma cell line Bel-7404]

OBJECTIVE: To investigate the anti-tumor effect of Paeonol (Pae) on the hepatocellular carcinoma cell line Bel-7404 and its molecular mechanisms. METHODS: Hepatocellular carcinoma cell line Bel-7404 was treated by Pae in various concentrations and different time points respectively; and then the cell proliferation was assayed by light microscope, MTT method. DNA agarose gel electrophoresis and TUNEL were used to detect the apoptosis. The expression of PTEN and Akt were examined by RT-PCR and immunocytochemical ABC method. RESULTS: Compared with the control groups Pae obviously increased the inhibitory and apoptosis rate of hepatocellular carcinoma cell line Bel-7404. It also showed a typical apoptotic morphology and DNA depicted a ladder pattern characteristic of the apoptosis, indicating the presence of DNA fragmentation. RT-PCR and immunocytochemical ABC assay showed that Pae could increase the expression of PTEN and decrease the expression of Akt. CONCLUSION: Pae can increase the anti-hepatocellular carcinoma effect, and its mechanism may be the increase of apoptosis-inducing effect which is regulated by phosphatidylinositol-3-kinase.     

芹菜素增敏TRAIL诱导卵巢癌CoC1细胞凋亡

目的研究芹菜素（API）抑制NF-κB活性,增强肿瘤坏死因子相关凋亡诱导配体（TRAIL）诱导人卵巢癌CoC1细胞凋亡的作用。方法体外培养CoC1细胞。碘化丙啶（PI）染色流式细胞术（FCM）定量分析细胞凋亡率（亚二倍体DNA含量细胞百分率）。DNA琼脂糖凝胶电泳观察细胞DNA梯形条带。Western blot检测细胞蛋白的表达。结果 API（20μmol/L）和TRAIL（20 ng/mL）以及两者合用48 h,CoC1细胞凋亡率分别是8.83%±2.33%、8.32%±2.80%和69.5%±4.65%;API（20μmol/L）预孵育4 h后,TRAIL（20 ng/mL）处理CoC1细胞44h,展示出典型DNA梯形条带图谱。API（20μmol/L）以时间依赖的方式降低CoC1细胞IκBα蛋白磷酸化水平和抑制NF-κB（p65）蛋白表达。结论亚细胞毒性浓度的API具有增强TRAIL诱导人卵巢癌CoC1细胞凋亡作用,其作用机制与抑制NF-κB活性有关。     

Growth inhibition and gene induction in human hepatocellular carcinoma cell exposed to sodium 4-phenylbutanoate

Background Sodium 4-phenylbutanoate （NaPB） can induce cellular differentiation and cell cycle arrest. However, its potential anticancer properties in hepatocellular carcinoma and influence on normal liver cell are still unclear. We observed the effects of NaPB on growth inhibition, including differentiation and phase growth arrest in normal liver cell line L-02 and hepatocellular carcinoma cell line Bel-7402. Furthermore, we investigated its mechanism in Bel-7402. Methods Hepatocellular carcinoma cells Bel-7402 and normal liver cell line L-02 were treated with NaPB at different concentrations. Light microscopy was used to find morphological change in cells. Cell cycle was detected by flow cytometry. Expression of acetylating histone H4 and of histones deacetylase 4 （HDAC4） were determined by Western blot. The expression of P21WAF1/CIP1 and E-cadherin were observed through immunocytochemistry. Results NaPB treatment led to time dependent growth inhibition in hepatocellular carcinoma cells Bel-7402. NaPB treatment caused a significant decline in the fraction of S phase cells and a significant increase in Go/G1 cells. NaPB increased the expression of P21wAFVCIP1 and E-cadherin in Bel-7402 and significantly decreased the level of HDAC4 in Bel-7402. NaPB significantly improved the level of acetylating histone H4. The normal liver cell line L-02 showed no distinct changes under treatment with NaPB. Conclusions NaPB inhibited the growth of hepatocellular carcinoma cells Bel-7402 and induced partial differentiation through enhancing the acetylating histones. In Bel-7402, the expressions of P21WAF1/CIP1 and E-cadherin may be related to level of acetylating histones and inhibition of cellular growth. NaPB showed no significant effect on normal liver cells.     

Growth Inhibition and Apoptosis Induction in Human Hepatoma Cells by Tanshinone Ⅱ_A

Summary:In order to study the effect of tanshinone Ⅱ_A on growth and apoptosis in human hepatomacell line BEL-7402 in vitro,the human hepatoma cell line BEL-7402 was treated with tanshinone Ⅱ_Aat various concentrations for 72 h.Growth suppression was evaluated by MTT assay;apoptosis-relat-ed alterations in morphology and biochemistry were ascertained under cytochemical staining(Hoechst33258),transmission electron microscopy(TEM),and DNA agarose gel electrophoresis.Apoptoticrate was quantified by flow cytometry(FCM).The results showed thst Tanshinone Ⅱ_A could inhibitthe growth of hepatoma cells in a dose-dependent manner,with IC_(50) value being 6.28μg/ml.Aftertreatment with 1—10 μg/ml tanshinone Ⅱ_A for 72 h,BEL-7402 cells apoptosis with nuclear chro-matin condensation and fragmentation as well as cell shrinkage and the formation of apoptotic bodieswere observed.DNA ladder could be demonstrated on DNA electrophoresis.FCM analysis showedhypodiploid peaks on histogram,and the apoptotic rates at 5