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1.
目的探讨siRNA沉默Fas死亡结构域相关蛋白(DAXX)基因对乳腺癌MCF-7细胞增殖和迁移行为的影响,为临床诊断提供参考依据。方法培养MCF-7细胞,观察组对细胞进行siRNA-DAXX转染,空白组不对细胞进行任何处理,对照组对细胞进行siRNA-NC转染,(1)使用RT-PCR和Western Blot法分别检测三组DAXX mRNA和蛋白表达水平,(2)使用MTT法检测三组细胞增殖能力,(3)使用Transwell小室实验检测三组细胞迁移能力。结果 (1)观察组DAXX表达水平与空白组及对照组对比差异有统计学意义(P<0. 05),siRNA能够显著抑制DAXX基因表达和蛋白合成,(2)Western blot显示DAXX基因被沉默后,观察组细胞增殖率仅为空白组或对照组的62. 93%,(3)RT-PCR显示DAXX基因被沉默后,观察组细胞增殖率仅为空白组或对照组的77. 24%;(4)Transwell小室实验结果显示,与空白组及对照组相比,观察组细胞穿过小室的几率显著降低,差异有统计学意义(P<0. 05)。结论 siRNA沉默DAXX基因表达后,乳腺癌MCF-7细胞的增殖能力和迁移能力显著减低,DAXX是一种肿瘤促进基因。  相似文献   

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目的:研究siRNA(small interfering RNA)抑制钙激活性中电导钾离子通道(intermediate-conductance Ca2+-activated K+channels,IKCa1)表达对宫颈癌HeLa细胞增殖的影响。方法:构建含有IKCa1基因的siRNA表达载体,脂质体法转染HeLa细胞,通过RT-PCR和Western Blot检测HeLa细胞IKCa1 mRNA和蛋白表达变化;WST-1检测细胞增殖能力;流式细胞仪分析HeLa细胞周期变化。结果:成功构建针对IKCa1基因的真核表达载体,所获表达载体转染HeLa细胞可使IKCa1mRNA和蛋白表达水平显著降低(P<0.05),并能有效抑制HeLa细胞增殖(P<0.05),使细胞周期停留在G0~G1期,S期细胞明显减少。结论:应用siRNA干扰技术能有效抑制IKCa1基因的表达,同时也可有效抑制宫颈癌HeLa细胞体外增殖活性。  相似文献   

3.
目的分析RNA干扰中性粒细胞相关载脂蛋白(NGAL)基因的表达对乳腺癌细胞增殖凋亡的影响及机制。方法利用蛋白质印迹(Western Blot)方法检测乳腺癌组织及人乳腺癌MCF-7、T47D和MDA-MB-231细胞株中NGAL的蛋白表达;小干扰RNA(siRNA)-NC及NGAL-siRNA转染人乳腺癌MCF-7细胞,未转染任何siRNA作为对照组,检测转染48 h后各组细胞中NGAL的蛋白表达情况;利用细胞计数试剂盒(CCK8)方法检测细胞的增殖情况;利用流式细胞仪检测细胞的凋亡;利用Western Blot方法检测活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、磷脂酰肌醇-3(PI3K)、丝氨酸苏氨酸激酶(AKT)及磷酸化的丝氨酸苏氨酸激酶(p-AKT)的蛋白表达情况。结果乳腺癌组织中NGAL的蛋白表达显著高于癌旁组织(P0.05),NGAL于乳腺癌MCF-7细胞中的蛋白表达最高,选择其作为后续研究;siRNA-NC组NGAL、Cleaved Caspase-3、PI3K及p-AKT蛋白表达和细胞存活率及凋亡率与相应的对照组比较差异无统计学意义(P0.05),而NGAL-siRNA组NGAL、PI3K和p-AKT蛋白表达及细胞存活率均显著低于相应的对照组(P0.05),NGAL-siRNA组的细胞凋亡率及Cleaved Caspase-3蛋白表达均显著高于相应的对照组(P0.05)。结论 NGAL基因在乳腺癌组织及细胞中高表达,抑制NGAL基因的表达会显著降低细胞增殖及促进细胞凋亡,其机制与PI3K/AKT信号通路的调控有关。  相似文献   

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目的 探讨RNA干扰HMGA1基因表达对神经胶质瘤细胞增殖凋亡的影响及机制。 方法 RT-PCR及Western blot检测神经胶质瘤组织中HMGA1的mRNA及蛋白表达;siRNA-NC、HMGA1-siRNA1、HMGA1-siRNA2转染人神经胶质瘤U251细胞,未转染任何siRNA作为空白对照组,48 h后检测各组细胞中HMGA1的蛋白表达;CCK8实验检测细胞增殖;流式细胞仪检测细胞凋亡;Western blot检测Ki67、PCNA、cleaved caspase-3、β-catenin、cyclin D1蛋白表达。 结果 神经胶质瘤组织中HMGA1基因的mRNA及蛋白表达均显著高于瘤旁组织(P<0.01);RNA干扰HMGA1基因能显著抑制其表达,HMGA1-siRNA2组的抑制效果更明显,选择作为后续研究;与对照组及siRNA-NC组比较,HMGA1-siRNA组细胞存活率及Ki67、PCNA、β-catenin、cyclin D1蛋白表达显著降低,细胞凋亡率及cleaved caspase-3蛋白表达显著升高(P<0.01)。 结论 HMGA1基因在神经胶质瘤组织中高表达,抑制其表达可通过下调Wnt/β-catenin信号通路抑制人神经胶质瘤U251细胞的增殖及促进凋亡。  相似文献   

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目的探讨抑制人滋养层细胞表面抗原-2(TROP2)基因表达对宫颈癌细胞凋亡的影响。方法空白对照组(Control组)、阴性对照无意义小干扰RNA siRNA序列组(Control siRNA组)和转染TROP2的靶向siRNA序列组(TROP2 siRNA组)转染人宫颈癌Hela细胞,48 h后蛋白印迹检测TROP2、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase3)、β-连环蛋白(β-catenin)、细胞周期素D1(Cyclin D1)蛋白表达;流式细胞仪检测细胞凋亡。结果转染TROP2-siRNA后能显著降低TROP2的蛋白表达;TROP2-siRNA组细胞凋亡率及Cleaved caspase3蛋白表达显著高于Control组,β-catenin、Cyclin D1蛋白表达显著低于Control组(P0.01),Control-siRNA组细胞凋亡率及Cleaved caspase3、β-catenin、Cyclin D1蛋白表达与Control组差异无统计学意义(P0.05)。结论抑制宫颈癌细胞中TROP2基因表达可促进癌细胞的凋亡,其机制与上调Cleaved caspase3蛋白表达及下调Wnt/β-catenin信号通路有关。  相似文献   

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目的 应用小干扰RNA(siRNA)干扰胰腺癌PANC-1细胞的c-Src表达,为后续试验做准备.方法 将c-Src siRNA,经LipofectamineTM2000脂质体转染胰腺癌PANC-1细胞后,用逆转录-聚合酶链反应(RT-PCR)和免疫印迹技术(Western blot)分别检测c-Src基因mRNA和c-Src蛋白表达的变化.结果 转染后胰腺癌PANC-1细胞的c-Src的mRNA与蛋白表达明显减少.结论 运用RNA干扰技术,可以有效地干扰胰腺癌PANC-1细胞c-Src的表达.  相似文献   

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目的 观察载体介导的RNA干扰技术能否有效抑制人乳腺癌细胞株MCF-7的hTERT的表达及其对细胞周期及凋亡的影响.方法 构建2个针对hTERT基因表达短发夹状siRNA的真核表达载体(pshRNA-hTERT)并转染人乳腺癌细胞株MCF-7,通过RT-PCR检测该基因mRNA,Western blot蛋白质检测,端粒酶活性检测,流式细胞仪检测细胞周期及凋亡等方法检测RNAi效果,从中筛选出基因沉默效果好的靶位点.结果 转染乳腺癌细胞后,pshRNA - hTERT1,2质粒均能抑制hTERT基因表达,mRNA表达分别下调了54.6%,55.2% 蛋白表达分别下调46.6%,47.8%.端粒酶活性均明显下降.对乳腺癌细胞周期中S期细胞明显减少,G1/G0期细胞显著增加.结论 RNAi在体外明显抑制乳腺癌细胞中hTERT基因的表达和瘤细胞增殖.  相似文献   

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目的探讨通过小干扰RNA(siRNA)抑制MICA基因表达对胃癌SGC-7901细胞生长的抑制作用。方法将靶向MICA的siRNA(MICA siRNA)通过脂质体转染法瞬时转染胃癌SGC-7901细胞48 h;Western blot检测siRNA转染前后SCG-7901细胞内MICA蛋白表达的变化;四甲基偶氮唑盐(MTT)法检测MICA siRNA转染对细胞增殖的影响,TUNEL检测siRNA转染对细胞凋亡的影响。结果 MICA siRNA转染干扰SCG-7901细胞内MICA表达,并诱导细胞凋亡和抑制细胞增殖。细胞转染后,MICA蛋白表达降低70%~85%,明显低于对照组,差异有统计学意义(P0.05);转染组细胞存活率明显低于对照组,差异有统计学意义(P0.05);转染组细胞凋亡率明显高于对照组,差异有统计学意义(P0.05)。结论靶向MICA抑制胃癌SGC-7901细胞增殖并促进细胞凋亡,MICA可用于胃癌基因治疗的靶点。  相似文献   

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目的:探讨RNA干扰技术对人卵巢癌细胞SKOV3中mTOR的表达及对细胞增殖与凋亡的影响。方法:培养SKOV3细胞系,设计合成mTOR siRNA实验分4组:正常培养组:未转染的正常培养SKOV3细胞;空白对照组:转染空脂质体的SKOV3细胞;转染组:转染mTOR siRNA的SKOV3细胞;无义对照组:转染无义siRNA的SKOV3细胞。采用Western blot法检测各组细胞中的mTOR蛋白表达水平;应用MTT法检测细胞增殖;流式细胞仪检测细胞凋亡。结果:mTOR siRNA转染组SK-OV3细胞mTOR蛋白的表达显著低于各对照组(P<0.05);mTOR siRNA转染组SKOV3细胞增殖低于各对照组(P<0.05);mTOR siRNA转染组细胞凋亡率高于各对照组(P<0.05)。结论:mTOR siRNA对SKOV3细胞中mTOR蛋白的表达有明显的抑制作用,特异性阻断mTOR基因表达可显著抑制SKOV3细胞的增殖并促进其凋亡。  相似文献   

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目的:探讨葡萄糖调节蛋白78(GRP78)在蛋白酶体抑制剂诱导卵巢癌细胞凋亡中的作用。方法:对3种卵巢癌细胞系SKOV3、A2870和CAOV3分别设空白对照组和不同蛋白酶体抑制剂处理组;对SKOV3细胞分别转染GRP78小干扰RNA(siRNA)和随机序列核酸siRNA组。利用蛋白酶体抑制剂MG132(5μmol/L)、PSI(20nmol/L)和EPOX(10nmol/L)作用3种细胞系,实时定量PCR和Western blot检测各组细胞GRP78 mRNA和蛋白表达。MTT和流式细胞术检测细胞活力和凋亡率。结果:各种蛋白酶体抑制剂均增加卵巢癌细胞中GRP78mRNA和蛋白表达水平。当GRP78siRNA靶向沉默GRP78表达后,蛋白酶体抑制剂作用后SKOV3细胞活力明显降低。结论:GRP78诱导表达干扰蛋白酶体抑制剂抗卵巢癌活性;沉默GRP78基因表达可提高蛋白酶体抑制剂对卵巢癌细胞的杀伤作用。  相似文献   

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Acute arsenic intoxication from environmental arsenic exposure   总被引:2,自引:0,他引:2  
Reports of acute arsenic poisoning arising from environmental exposure are rare. Two cases of acute arsenic intoxication resulting from ingestion of contaminated well water are described. These patients experienced a variety of problems: acute gastrointestinal symptoms, central and peripheral neurotoxicity, bone marrow suppression, hepatic toxicity, and mild mucous membrane and cutaneous changes. Although located adjacent to an abandoned mine, the well water had been tested for microorganisms only and was found to be "safe." Regulations for testing of water from private wells for fitness to drink are frequently nonexistent, or only mandate biologic tests for microorganisms. Well water, particularly in areas near mining activity, should be tested for metals.  相似文献   

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地方性砷中毒是一种严重危害病区居民健康的地方病.根据砷源不同,分为饮水型职业性砷中毒、燃煤型职业性砷中毒和职业性砷中毒.饮水型地方性砷中毒主要是由于长期暴露于饮用水中的砷而引起的慢性砷中毒.饮水中的砷还可以通过农田灌溉等引起粮食、土壤等砷浓度的升高,间接危害居民健康.在以往的研究中发现,饮水型砷中毒地区饮用水砷超标,不...  相似文献   

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长期吸入砷化合物可发生慢性砷中毒,发生早晚及程度与车间空气砷浓度大致呈平行关系。不脱离砷接触时尿砷可作为砷中毒诊断的重要指标,对尿砷超标者行驱砷治疗可降低砷中毒发病率。  相似文献   

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The large disparity between arsenic concentrations in drinking water and urine remains unexplained. This study aims to evaluate predictors of urinary arsenic in a population exposed to low concentrations (≤50?μg/l) of arsenic in drinking water. Urine and drinking water samples were collected from a subsample (n=343) of a population enrolled in a bladder cancer case-control study in southeastern Michigan. Total arsenic in water and arsenic species in urine were determined using ICP-MS: arsenobetaine (AsB), arsenite (As[III]), arsenate (As[V]), methylarsenic acid (MMA[V]), and dimethylarsenic acid (DMA[V]). The sum of As[III], As[V], MMA[V], and DMA[V] was denoted as SumAs. Dietary information was obtained through a self-reported food intake questionnaire. Log(10)-transformed drinking water arsenic concentration at home was a significant (P<0.0001) predictor of SumAs (R(2)=0.18). Associations improved (R(2)=0.29, P<0.0001) when individuals with less than 1?μg/l of arsenic in drinking water were removed and further improved when analyses were applied to individuals who consumed amounts of home drinking water above the median volume (R(2)=0.40, P<0.0001). A separate analysis indicated that AsB and DMA[V] were significantly correlated with fish and shellfish consumption, which may suggest that seafood intake influences DMA[V] excretion. The Spearman correlation between arsenic concentration in toenails and SumAs was 0.36 and between arsenic concentration in toenails and arsenic concentration in water was 0.42. Results show that arsenic exposure from drinking water consumption is an important determinant of urinary arsenic concentrations, even in a population exposed to relatively low levels of arsenic in drinking water, and suggest that seafood intake may influence urinary DMA[V] concentrations.  相似文献   

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The Binational Arsenic Exposure Survey (BAsES) was designed to evaluate probable arsenic exposures in selected areas of southern Arizona and northern Mexico, two regions with known elevated levels of arsenic in groundwater reserves. This paper describes the methodology of BAsES and the relationship between estimated arsenic intake from beverages and arsenic output in urine. Households from eight communities were selected for their varying groundwater arsenic concentrations in Arizona, USA and Sonora, Mexico. Adults responded to questionnaires and provided dietary information. A first morning urine void and water from all household drinking sources were collected. Associations between urinary arsenic concentration (total, organic, inorganic) and estimated level of arsenic consumed from water and other beverages were evaluated through crude associations and by random effects models. Median estimated total arsenic intake from beverages among participants from Arizona communities ranged from 1.7 to 14.1 μg/day compared to 0.6 to 3.4 μg/day among those from Mexico communities. In contrast, median urinary inorganic arsenic concentrations were greatest among participants from Hermosillo, Mexico (6.2 μg/L) whereas a high of 2.0 μg/L was found among participants from Ajo, Arizona. Estimated arsenic intake from drinking water was associated with urinary total arsenic concentration (p < 0.001), urinary inorganic arsenic concentration (p < 0.001), and urinary sum of species (p < 0.001). Urinary arsenic concentrations increased between 7% and 12% for each one percent increase in arsenic consumed from drinking water. Variability in arsenic intake from beverages and urinary arsenic output yielded counter intuitive results. Estimated intake of arsenic from all beverages was greatest among Arizonans yet participants in Mexico had higher urinary total and inorganic arsenic concentrations. Other contributors to urinary arsenic concentrations should be evaluated.  相似文献   

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尚琪  任修勤  李晋蓉 《卫生研究》2002,31(4):270-272
利用人群流行病学调查资料 ,通过估计人群累积呼吸量和大米食用量的方法 ,估算了污染区人群经呼吸和饮食的累积砷暴露量。并对如何评价人群环境污染物暴露水平的方法进行了探讨。不考虑非污染期经呼吸道的砷暴露时 ,所估计的污染区人群最大累积砷暴露量 ,经呼吸道途径 ,女性为 5 91 4mg ,男性为6 12 9mg。消化道途径为 34 88 74mg,合计女性为 40 80 14mg ,男性为 410 1 6 6mg。按污染时间折算成每日暴露量 :女性为 34 9 1μg (天·人 ) ,男性为 35 0 9μg (天·人 ) ,约为调查时所计算日均暴露量的 80 %左右  相似文献   

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