Genistein对bFGF诱导的EJ细胞增殖的抑制作用

目的：观察碱性成纤维细胞生长因子（bFGF）对人膀胱癌细胞株EJ细胞的增殖作用;以及酪氨酸蛋白激酶（TPK）抑制剂Genistein对bFGF诱导的EJ细胞增殖的抑制作用。方法：以不同浓度的bFGF刺激EJ细胞,再对由bFGF引起的细胞增殖施加不同浓度的Genistein;以MTT法检测细胞的增殖程度,原位凋亡细胞检测（TUNEL）和流式细胞仪（FCM）检测Genistein对EJ细胞凋亡和细胞周期的影响。结果：bFGF可以使EJ细胞增殖比明显上升,Genistein可引起细胞增殖比明显下降,其程度均随浓度增高而增强;Genistein可以促进EJ细胞的凋亡,使细胞阻滞于G2/M期。结论：bFGF可以诱导EJ细胞的增殖,Genistein可以诱导EJ细胞凋亡,对bFGF诱导的细胞增殖有抑制作用,可能为膀胱肿瘤的治疗提供新的方法。     

重组腺病毒载体CRT联合紫杉醇对乳腺癌细胞周期、增殖及凋亡的影响

目的：研究Ad-CRT联合紫杉醇对乳腺癌MDA-MB-231细胞周期、细胞增殖及凋亡的影响。方法：MTT法检测细胞增殖,相差显微镜观察细胞形态学变化;PI单染流式细胞术检测细胞周期和凋亡。结果：Ad-CRT联合紫杉醇对乳腺癌细胞MDA-MB-231增殖的抑制率明显高于单纯Ad-CRT组和单纯紫杉醇组,且抑制作用呈时间依赖关系（P〈0.05）,且细胞形态发生明显的改变。Ad-CRT联合紫杉醇将乳腺癌细胞MDA-MB-231的细胞周期明显的阻滞在G2/M期,并且促进细胞的凋亡。结论：Ad-CRT联合紫杉醇能增强乳腺癌细胞MDA-MB-231增殖抑制、阻滞细胞周期并且诱导细胞凋亡。     

Genistein对bFGF诱导的EJ细胞增殖的抑制作用

目的:观察碱性成纤维细胞生长因子(bFGF)对人膀胱癌细胞株EJ细胞的增殖作用;以及酪氨酸蛋白激酶(TPK)抑制剂Genistein对bFGF诱导的EJ细胞增殖的抑制作用。方法:以不同浓度的bFGF刺激EJ细胞,再对由bFGF引起的细胞增殖施加不同浓度的Genistein;以MTT法检测细胞的增殖程度,原位凋亡细胞检测(TUNEL)和流式细胞仪(FCM)检测Genistein对EJ细胞凋亡和细胞周期的影响。结果:bFGF可以使EJ细胞增殖比明显上升,Genistein可引起细胞增殖比明显下降,其程度均随浓度增高而增强;Genistein可以促进EJ细胞的凋亡,使细胞阻滞于G2/M期。结论:bFGF可以诱导EJ细胞的增殖,Genistein可以诱导EJ细胞凋亡,对bFGF诱导的细胞增殖有抑制作用,可能为膀胱肿瘤的治疗提供新的方法。     

Genistein抑制人卵巢癌细胞系SKOV3增殖和诱导凋亡发生的研究

背景与目的:许多研究表明 Genistein对多种肿瘤细胞有抑制作用,但有关 Genistein对卵巢癌细胞作用的报道很少.本研究旨在通过观测 Genistein对人卵巢癌细胞系 SKOV3的抑制增殖和诱导凋亡作用,探讨其抗癌作用的机理.方法:应用四甲基偶氮唑盐( MTT)法检测不同浓度 Genistein对 SKOV3的生长抑制作用;吖啶橙 /溴乙锭( AO/EB)荧光染色法及电镜观察凋亡细胞及凋亡小体;流式细胞仪分析细胞周期及凋亡率;琼脂糖凝胶电泳检测凋亡特征性 DNA梯形带;免疫细胞化学法及 RT- PCR法分别检测细胞增殖凋亡调控相关蛋白及其 mRNA的表达。结果： Genistein对 SKOV3细胞的增殖抑制作用呈时间及浓度依赖性, 20 μ mol/L和 40 μ mol/L Genistein作用 72 h后,细胞的生长抑制率达 72. 07％和 74. 93％。 20 μ mol/L Genistein作用 SKOV3细胞 48 h后出现细胞周期的 G2/M期阻滞。荧光显微镜和电镜均观察到用药后凋亡细胞典型的形态学特征。细胞凋亡率以 Genistein 20 μ mol/L组最高,达 23. 7％。凝胶电泳观察到特征性 DNA梯形带。 20 μ mol/L Genistein作用 SKOV3 48 h后, bcl- 2基因表达水平降低,而 p21WAF1/CIP1和 bax基因表达增加（ P< 0. 05）; PCNA、Bcl- 2及 cyclin B1蛋白表达水平降低, Bax、p21WAF1/CIP1蛋白表达增加（ P< 0. 01）。结论： Genistein可抑制卵巢癌细胞系 SKOV3的增殖并诱导其发生凋亡, Genistein可能通过上调 p21WAF1/CIP1基因及蛋白水平、下调 cyclin B1及 PCNA蛋白表达水平抑制增殖;通过下调 bcl- 2基因及蛋白表达、上调 bax基因及蛋白表达诱导卵巢癌细胞凋亡.     

Genistein对耐药卵巢癌细胞SKOV-3增殖、凋亡和顺铂敏感性的影响

目的探讨Genistein对耐药卵巢癌细胞SKOV3增殖、凋亡和顺铂敏感性的影响。方法采用MTT法检测Genistein单用及合用顺铂对细胞增殖的影响,流式细胞仪分析Genistein及Genistein联合顺铂对细胞周期和凋亡的影响。结果Genistein对SKOV3细胞增殖表现出浓度依赖性的抑制作用,并显著提高了其对顺铂的敏感性(P<0.05);0.625～5μg/ml顺铂与2.5～10μg/mlGenistein合用基本表现为协同作用。5、10μg/mlGenistein均可将细胞阻滞于G2/M期并诱导一定程度的凋亡,10μg/mlGenistein还可进一步的干扰S期进程;当顺铂与Genistein合用时,两种药物在干扰SKOV3细胞周期和诱导凋亡方面表现为显著的协同效应。结论Genistein能够抑制耐药卵巢癌细胞SKOV3的增殖,并显著增强该细胞对顺铂的敏感性。     

马鞭草C部位诱导人绒毛膜癌JAR细胞凋亡分子机制研究

目的:探讨马鞭草C部位诱导人绒毛膜癌JAR细胞凋亡的分子机制.方法:流式细胞仪检测细胞周期;Western-blot及RT-PCR检测Fas/Fasl蛋白及基因的表达.结果:药物作用后细胞被不同程度阻滞于G2/M期,并伴有G0/G1期比例下降;Fasl蛋白表达降低,Fas在C部位作用前后均无表达;Fasl mRNA转录呈下降趋势.结论:马鞭草C部位将人绒毛膜癌JAR细胞阻滞在G2/M期,并诱导其凋亡,其作用机制可能与Fasl的表达下调有关.     

大蒜素对骨肉瘤MG-63细胞系增殖和凋亡的影响

目的观察大蒜素对人成骨肉瘤细胞株MG-63增殖和凋亡的影响。方法不同浓度的大蒜素作用于体外培养的MG-63细胞,倒置显微镜下观察MG-63细胞的形态学变化,采用CCK-8法检测大蒜素对MG-63细胞增殖抑制能力,吖啶橙/溴化乙锭双荧光染色及Annexin V-FITC标记流式细胞术检测细胞凋亡情况以及对MG-63细胞周期影响。结果大蒜素可抑制人成骨肉瘤细胞株MG-63细胞增殖,具有明显剂量和时间依赖性,10 μg/ml大蒜素可明显诱导MG-63细胞凋亡,并将细胞周期阻滞在G2/M期。结论大蒜素能够诱导MG-63细胞凋亡,阻滞细胞周期,抑制细胞增殖。     

三氧化二砷对胃癌BGC-823细胞增殖及细胞周期的影响

[目的]探讨三氧化二砷（AS2O3）对胃癌BGC-823细胞增殖的抑制作用及对细胞周期的影响。[方法]应用MTT方法检测三氧化二砷对胃癌BGC-823细胞增殖的抑制作用，流式细胞仪进行细胞周期解析及凋亡检测．[结果]三氧化二砷对胃癌BGC823细胞具有杀伤作用，且呈时间-浓度依赖性抑制肿瘤细胞的增殖．72h抑制细胞50％增殖的药物浓度约为3．3μmol／L，与对照组比差异显著（P=0．0048）；5μmol／LAs2O3作用24h，细胞形态学可见典型的凋亡细胞及M期阻滞；流式细胞仪进行细胞周期解析结果提示，G2/M期细胞由正常对照的25％增加到46．3％，出现了明显的G2/M期阻滞，亚二倍体凋亡细胞由4％增加到18％。[结论]三氧化二砷抑制胃癌细胞的增殖．诱导细胞周期阻滞及凋亡。     

三氧化二砷对胃癌BGC-823细胞增殖及细胞周期的影响

[目的]探讨三氧化二砷（AS2O3）对胃癌BGC-823细胞增殖的抑制作用及对细胞周期的影响。[方法]应用MTT方法检测三氧化二砷对胃癌BGC-823细胞增殖的抑制作用，流式细胞仪进行细胞周期解析及凋亡检测．[结果]三氧化二砷对胃癌BGC823细胞具有杀伤作用，且呈时间-浓度依赖性抑制肿瘤细胞的增殖．72h抑制细胞50％增殖的药物浓度约为3．3μmol／L，与对照组比差异显著（P=0．0048）；5μmol／LAs2O3作用24h，细胞形态学可见典型的凋亡细胞及M期阻滞；流式细胞仪进行细胞周期解析结果提示，G2/M期细胞由正常对照的25％增加到46．3％，出现了明显的G2/M期阻滞，亚二倍体凋亡细胞由4％增加到18％。[结论]三氧化二砷抑制胃癌细胞的增殖．诱导细胞周期阻滞及凋亡。     

马鞭草C部位单体４′-甲醚-黄芩素对人绒毛膜癌细胞增殖的抑制作用

摘 要 目的: 探讨马鞭草有效成分所提取的单体4′-甲醚-黄芩素(4′methyletherscutellarein，4MS)对人绒毛膜癌JAR细胞的增殖抑制作用及其相关机制。方法：JAR细胞传代后加入不同浓度4MS作用一定时间，应用MTT法检测对细胞增殖的抑制作用,流式细胞术测定细胞凋亡率及细胞周期变化, AO/EB双染法区分早、晚期凋亡细胞和坏死细胞，RTPCR技术分析4MS对人绒毛膜癌JAR细胞凋亡相关基因Caspase 3、p38 MAPK及Survivin表达的影响, 并进一步用Western blotting测定Caspase 3、p38 MAPK及Survivin在用药前后蛋白水平的表达差异。结果：不同质量浓度(10、20、40 mg/L)的4MS对JAR细胞均有增殖抑制作用,并随药物浓度和作用时间的增加而不断增强(P<0.05，P<0.01); 流式细胞术显示,随着药物浓度的增加细胞凋亡率亦逐渐升高,G2/M期细胞所占比例增大 (P<0.05); AO/EB双染后发现，随着4MS剂量的增加，晚期凋亡细胞逐渐增多; 20和40 mg/L的4MS作用48 h后, JAR细胞中p38 MAPK及Caspase 3 mRNA表达下降，Survivin mRNA表达上升，与对照组相比均有统计学意义(P<0.05)；Western blotting证实，20 mg/L和40 mg/L 4MS作用48 h后Survivin蛋白表达量显著下降，p38磷酸化水平升高，Caspase 3被明显激活。 结论： 4MS能抑制人绒毛膜癌JAR细胞的增殖并诱导凋亡，可能与其阻滞细胞生长于G2/M期、抑制Survivin抗凋亡活性，直接激活p38 MAPK信号通路和Caspase 3活化有关。     

吴茱萸碱联合射线对舌鳞状细胞癌Tca-8113细胞增殖、黏附和迁移的影响

目的:探讨吴茱萸碱(evodiamine,EVO)联合射线对舌鳞状细胞癌Tca-8113细胞放射增敏、黏附和迁移的影响。方法:采用MTT比色法检测EVO对Tca-8113细胞增殖及放射增敏的影响;克隆形成实验检测EVO联合射线对Tca-8113细胞克隆形成能力的影响;细胞黏附和划痕实验检测EVO联合射线作用后细胞黏附和迁移能力的改变。结果:Tca-8113细胞的存活率随EVO作用浓度及作用时间的增加而降低,而不同放射剂量和处理不同时间后EVO+放射组的细胞存活率均显著低于单纯放射组(P<0.01)。克隆形成实验结果显示,EVO的放射增敏比(sensitizing enhancement ratio,SER)Do=1.35,SERDq=1.75,SERSF2=1.67。同质黏附实验显示,20、40和60min时单纯放射组及EVO+放射组与对照组相比,黏附细胞数显著增加(P<0.01);且在40和60min时,EVO+放射组与单纯放射组相比,黏附细胞数亦显著增加(P<0.01)。与血管内皮细胞的黏附实验显示,在20、40和60min时,EVO+放射组与单纯放射组及对照组相比,黏附率均明显下降(P<0.05)。划痕实验结果显示,EVO联合放射组的细胞迁移率显著低于对照组和单纯放射组(P<0.01)。结论:EVO对Tca-8113细胞有明显的放射增敏作用,并能改变Tca-8113细胞的黏附和迁移能力。     

苦参碱对人类肝癌细胞HepG2增殖、细胞周期及凋亡的影响

 目的　探讨苦参碱（MT）对人类肝癌细胞株HepG2增殖、细胞周期及凋亡的影响。方法　用不同质量浓度的MT（0.0125、0.025、0.05、0.1、0.2、0.4、0.8、1.6 g／L）对培养的HepG2细胞作用不同时间（24、48、72 h）,用四氮噻唑蓝（MTT） 比色法检测MT对细胞增殖的影响;流式细胞术（FCM）检测MT对HepG2细胞周期及凋亡的影响。结果　质量浓度≥0.1 g／L的MT有抑制HepG2细胞增殖的作用,且作用随药物质量浓度的增加及时间的延长而加强（P＜0.01）。FCM检测分析显示,0.8 g／L MT作用48 h能将HepG2细胞阻滞在G1期[（75.3±6.5）%对（64.1±6.3）%]（P＜0.05）,1.6 g／L MT作用48 h能将HepG2细胞阻滞在G2期[（29.1±9.1）%对（11.6±2.1）%]（P＜0.01）。0.4、0.8、1.6 g／L MT作用12、24、48 h对HepG2细胞都有诱导凋亡作用,且作用随药物质量浓度的增加及作用时间的延长而加强（P＜0.01）。结论　MT能够抑制HepG2细胞的增殖、诱导其凋亡并影响其细胞周期,呈时间和剂量依赖性。MT抗肝癌的机制可能与影响肝癌细胞周期、抑制细胞增殖及诱导凋亡有关。     

Micromolar taxol, with or without hyperthermia, induces mitotic catastrophe and cell necrosis in HeLa cells

Purpose: Although the mode of action of taxol, when used in nanomolar or micromolar concentrations during long periods, is extensively studied, there are few data available on taxol-mediated cytotoxicity when the drug is applied for a short time alone or in combination with hyperthermia. We studied the effect of taxol and hyperthermia on cell cycle kinetics, proliferation, and mode of cell death in human cervical carcinoma HeLa cells, following a scheme which resembles the one currently used in regional chemotherapy. methods: Cells were incubated with micromolar doses of taxol for two h under normothermic or hyperthermic conditions and then cultured in drug-free medium for several days. Cell viability was assessed via an MTT assay. Necrotic and apoptotic cell death was determined using Trypan blue staining and TUNNEL assay, respectively. Flow cytometry was used for the analysis of cell cycle kinetics and the counting of apoptotic cells. Mitotic index, nuclear morphology and nuclear envelope organization were analyzed by fluorescence microscopy. results: Cells exposed to micromolar doses of taxol for 2 h and then transferred to a drug-free medium for 24 h were arrested at G2/M or M phase. When treated cells were cultured in normal media for longer periods, most of them remained in a tetraploid state, became multinucleated without properly completing cytokinesis and died mostly by necrosis. Hyperthermia alone exerted a cytotoxic effect, inhibited proliferation and caused minor changes in cell cycle kinetics. When combined with taxol treatment, hyperthermia modified the cell cycle-arresting effects of the drug, but did not alter significantly taxol-mediated cytotoxicity. conclusions: From these data we conclude that short time incubation of HeLa cells under normothermic or hyperthermic conditions with micromolar concentrations of taxol is sufficient enough to induce extended cell growth arrest and cell death by necrosis.     

热疗联合紫杉醇对人舌鳞癌细胞系CAL-27增殖、凋亡和周期影响

目的:观察热疗联合紫杉醇对人舌鳞癌细胞系CAL-27增殖、凋亡和周期的影响并探讨其机制。方法:CCK-8法确定紫杉醇工作浓度,将体外培养CAL-27细胞分为对照组、紫杉醇组、42℃热疗组和联合治疗组。克隆形成实验检测细胞增殖能力,流式细胞术检测细胞周期及凋亡,蛋白印迹法检测AKT、p-AKT、Bcl-2、Bax蛋白表达...     

Multicompartment analysis of cell proliferation and cell migration in the Sezary syndrome

Serial radioautographic data obtained in two patients with Sezary syndrome following intravenous tritiated thymidine administration were analysed using multicompartment kinetic models. In both patients, the grain count halving time in the cutaneous Sezary cell compartment was too long to account for the grain count halving rate observed in the peripheral blood. This implies that some proliferating cell compartment other than the skin is primarily responsible for producing the Sezary cells that circulate in the peripheral blood. In both patients, the cell compartment that served as the source of circulating Sezary cells consisted of 5-6 X 10(11) cells (500-600 g of tumor) with an average cell cycle time of 3-4 days. By comparison, the cutaneous Sezary cell compartment was estimated to contain 2.2-4.6 X 10(12) cells (2.2-4.6 kg of tumour) with an average cell cycle time of 7-70 days. While this study does not permit direct anatomic localization of the primary site of Sezary cell production, the properties of this compartment that can be deduced from the available data suggest the lymph nodes as likely candidates. Thus, the Sezary syndrome may well be a true lymph node malignancy with prominent cutaneous manifestations.     

MT-3和MT-1E基因转染对食管癌细胞株增殖、细胞周期及凋亡的影响

背景与目的:金属硫蛋白(metallothionein,MT)-3具有细胞生长抑制活性,是目前已知唯一的具有独特生理功能的MT亚型.MT-1E是MT-1的1个亚型,有文献报道MT-1蛋白的表达量与食管癌的恶性程度呈正相关.本研究拟探讨MT-3和MT-lE基因转染对食管癌细胞株增殖、细胞周期及凋亡的影响.方法:采用阳离子脂质体法,将己构建好的MT-3,MT-1E及其空载体4种质粒转染食管癌Eca-109和TE13细胞株:荧光显微镜判断转染效率,RT-PCR检测目的基因的表达,四甲基偶氮唑蓝(MTT)法检测细胞增殖能力,流式细胞术检测细胞周期与细胞凋亡.结果:转染MT-3和MT-lE基因的食管癌细胞株均有外源目的基因相应mRNA的高表达.转染MT-3基因的Eca-109和TE13细胞株,其增殖能力受到明显抑制(P<0.05),Go/G,期细胞比例明显升高(PO.05).结论:上调MT-3基因表达能抑制食管癌细胞增殖,促进其凋亡,这可能是通过阻滞肿瘤细胞的生长周期来实现的.而MT-1 E基因可能在食管癌细胞的增殖过程中作用不明显.     

Carboxypeptidase E promotes cancer cell survival,but inhibits migration and invasion

Carboxypeptidase E (CPE), a prohormone processing enzyme is highly expressed and secreted from (neuro)endocrine tumors and gliomas, and has been implicated in cancer progression by promoting tumor growth. Our study demonstrates that secreted or exogenously applied CPE promotes survival of pheochromocytoma (PC12) and hepatocellular carcinoma (MHCC97H) cells under nutrient starvation and hypoxic conditions, but had no effect on their proliferation. CPE also reduced migration and invasion of fibrosarcoma (HT1080) cells. We show that CPE treatment mediates survival of MHCC97H cells during metabolic stress by up-regulating the expression of anti-apoptotic protein BCL-2, and other pro-survival genes, via activation of the ERK1/2 pathway.     

下调miR-21表达抑制鼻咽癌CNE2细胞增殖和侵袭

背景与目的：miR-21在人类多种肿瘤中异常高表达。本研究探讨干扰miR-21表达对鼻咽癌CNE2细胞增殖、迁移和侵袭的影响。方法：使用脂质体将miR-21 inhibitor转染CNE2细胞,以无关序列(NC inhibitor)作为阴性对照。采用qRT-PCR技术验证miR-21 inhibitor转染的CNE2细胞中miR-21的表达水平;通过MTS法、细胞划痕、Transwell侵袭实验观察下调miR-21表达对CNE2细胞增殖、迁移和侵袭的影响。结果：转染miR-21 inhibitor的CNE2细胞中miR-21的表达明显下调,并且呈浓度依赖性。表明转染miR-21 inhibitor能有效抑制CNE2细胞中miR-21的表达。转染miR-21 inhibitor的CNE2细胞与对照细胞相比,增殖速度明显减慢,差异有统计学意义(P<0.05)。细胞划痕实验显示,下调miR-21表达抑制CNE2细胞迁移(P<0.05)。Transwell侵袭实验结果显示,下调miR-21表达抑制CNE2细胞侵袭(P<0.05)。结论：miR-21能促进鼻咽癌细胞增殖、迁移和侵袭,其可能在鼻咽癌的发生、发展中发挥重要作用。     

OTX008, a selective small-molecule inhibitor of galectin-1, downregulates cancer cell proliferation,invasion and tumour angiogenesis

BackgroundGalectin-1 (Gal1), a carbohydrate-binding protein is implicated in cancer cell proliferation, invasion and tumour angiogenesis. Several Gal1-targeting compounds have recently emerged. OTX008 is a calixarene derivative designed to bind the Gal1 amphipathic β-sheet conformation. Our study contributes to the current understanding of the role of Gal1 in cancer progression, providing mechanistic insights into the anti-tumoural activity of a novel small molecule Gal1-inhibitor.MethodsWe evaluated in vitro OTX008 effects in a panel of human cancer cell lines. For in vivo studies, an ovarian xenograft model was employed to analyse the antitumour activity. Finally, combination studies were performed to analyse potential synergistic effects of OTX008.ResultsIn cultured cancer cells, OTX008 inhibited proliferation and invasion at micromolar concentrations. Antiproliferative effects correlated with Gal1 expression across a large panel of cell lines. Furthermore, cell lines expressing epithelial differentiation markers were more sensitive than mesenchymal cells to OTX008. In SQ20B and A2780-1A9 cells, OTX008 inhibited Gal1 expression and ERK1/2 and AKT-dependent survival pathways, and induced G2/M cell cycle arrest through CDK1. OTX008 enhanced the antiproliferative effects of Semaphorin-3A (Sema3A) in SQ20B cells and reversed invasion induced by exogenous Gal1. In vivo, OTX008 inhibited growth of A2780-1A9 xenografts. OTX008 treatment was associated with downregulation of Gal1 and Ki67 in treated tumours, as well as decreased microvessel density and VEGFR2 expression. Finally, combination studies showed OTX008 synergy with several cytotoxic and targeted therapies, principally when OTX008 was administered first.ConclusionThis study provides insights into the role of Gal1 in cancer progression as well as OTX008 mechanism of action, and supports its further development as an anticancer agent.     

苦参碱对K562细胞株端粒酶活性和细胞周期的影响

目的为寻找新的肿瘤细胞诱导分化剂,探讨了苦参碱对Ｋ５６２细胞的诱导分化作用及其机制。方法采用ＰＣＲＥｌｉｓａ方法检测苦参碱作用前后端粒酶活性的改变,以流示细胞仪分析细胞周期的改变。结果苦参碱作用后的Ｋ５６２细胞,其端粒酶活性明显受抑,且细胞周期明显改变,Ｓ期细胞数显著降低。结论苦参碱对Ｋ５６２细胞有诱导分化作用,其机制可能与抑制端粒酶活性、阻止细胞周期的进程有关。