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目的 探讨马钱子碱抗肝细胞癌的作用及其机制.方法 体外培养人肝癌SMMC-7721细胞,加入不同浓度的马钱子碱(2.5~400μg/ml),细胞培养72 h,MTT法测定细胞生长抑制率.Western blotting和荧光定量RT-PCR技术分别测定培养24、48、72 h肝癌细胞PCNA、Cyclin D1、FAS基因mRNA和蛋白表达.结果 随着马钱子碱用量逐渐增加,对人肝癌细胞SMMC-7721生长抑制作用增强,马钱子碱用量为320 μg/ml时对肝癌细胞生长抑制率接近100%.马钱子碱作用肝癌细胞24 h与作用48 h相比,Fas蛋白和mRNA表达差异无统计学意义(分别F=2.547,1.582,均P>0.05),作用72 h时差异有统计学意义(分别F=1.036,1.137,均P<0.05);PCNA和Cyclin D1的mRNA和蛋白表达各时间点差异无统计学意义(PCNA分别F=3.612,2.174,3.029;Cyclin D1分别F=2.361,2.915,1.976,均P>0.05).结论 马钱子碱抑制肝癌细胞生长,可能通过肝癌细胞FAS基因和蛋白表达增加,诱导肝癌细胞凋亡发挥抑制作用,而与PCNA和Cyclin D1作用无关.Abstract: Objective To study the effect of brucine on the growth of a hepatocellular carcinoma cell line in vitro. Methods Brucine was added into a liver cancer cell line of SMMC-7721 in vitro, at drug concentration of brucine from 2. 5 μg/ml to 400 μg/ml. The inhibition rate of cell growth was measured by MTT technique after the cells were cultured for 72 hours. The protein and mRNA expression of PCNA,cyclin D1 and FAS were respectively assayed with Western blotting and fluorescent quantitation RT-PCR techniques at 24, 48, 72 h. Results The inhibition rate of liver cancer cell was near 100% when the brucine concentration was at 320 μg/ml. The protein and mRNA expression of FAS were of no significant difference at 24 h vs 48 h ( seperately F = 2. 547,1. 582, all P > 0. 05 ), and significant difference existed at 24 h vs 72 h( seperately F = 1. 036, 1. 137, all P < 0. 05 ). The protein and mRNA expression of PCNA,Cyclin D1 were of no significant difference between various time period( seperately PCNA F = 3.612,2. 174,3.029;Cyclin D1 F=2.361,2.915,1.976,all P>0.05). Conclusions Brucine inhibits the growth of liver cancer cells, by inducing increased apoptosis of the cells probably through FAS overexpression. 相似文献
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Objective To study the expression of aquaporin 1 ( AQP1 ) in pancreas and its pathogenic role in rats with experimental acute necrotizing pancreatitis (ANP). Methods Forty-eight male Sprague-Dawley rats were randomly divided into two groups : control group ( n = 24 ) and ANP group ( n = 24). Six rats in each group were sacrificed at 3,6,12 and 18 h after induction of experimental models. Quantity of ascites and levels of serum amylases were measured at each time point. Serum AQPI was determined by ELISA. Pathological changes of pancreatic tissues were examined. Capillary permeability in pancreatic tissues was tested by Evans Blue (EB) extravasation method. AQPI expression in pancreatic tissues was detected by immunohistochemieal staining, Western blot and fluorescence quantitative polymerase chain reaction (FQ-PCR). Results (1) Serum amylase level at 3,6,12, and 18 h in control group was (1308±759) ,(1077±508), (1325±761),(1328±762) U/L,and that in ANP group was (9102± 2199), (8799±1634), (9398±1473), (9484±862) U/L respectively. The concentration of EB in pancreatic tissues at each time point in control group was (205.61±32.99), (141.46±27.18 ), (96.94± 26.79), (61.43±24.82) mg/L, and that in ANP group was ( 273.59±23.47 ), ( 253.51±31.68 ), (221.15±73.68 ), (185.28±42.35) ,respectively. There was significandy difference between two groups. Serum AQP1 level in control group at each point was ( 74.08±11.80), (78.49±9.06 ), (75.77± 7.37 ), ( 72.75±13.87 ) mg/L, and that in AN P group was (73.29±9.61 ), ( 62.85±7.28 ), (62.07± 4.39 ), (46.33±11.91 ) mg/L, respectively ( P < 0.01 ). ( 2 ) Protein expression of AQPI in pancreatic tissues detected by immunohistochemical staining in control group at each time point was 114.13±7.92, 122.39±7.99,145.98±6.48,113.98±6.48, and that in ANP group was 80.07±14.89,110.54± 4.45,103.77±10.48,99.18±6.95;Protein expression of AQP1 in pancreatic tissues detected by Western blot in control group at each time point was 1.19±0.33,1.02±0.25,0.90±0.33,1.06±0.20,and that in ANP group was 0.83±0.11,0.96±0.21,0. 58±0.28,0.72±0.14, respectively ( P < 0.05 ). ( 3 ) The mRNA level of AQP1 gene expressed in pancreas in control group at each time point was 0.91±0.22,1.01±0.83,0.48±0.23,0.61±0.51,and that in ANP group was 2.13±0.63,2.02±1.40, 2.07±0.86,2.49±2.47,respectively (P<0.01).Condusion The expression of AQP1 wasdown-reg-ulated in pancreas in rats with ANP,which might could play an important role in the pathogenesis of capil-lary leak syndrome. 相似文献
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目的探讨胃肠道神经束膜瘤的临床病理特征。方法分析4例胃肠道神经束膜瘤的临床资料,总结其临床病理和免疫表型。结果 1例位于胃体大弯,1例位于结肠,2例位于直肠;瘤体直径0.3~1 cm,其特征是黏膜固有层内见温和的梭形细胞增生,表达神经束膜标志物,梭形细胞呈层状、束状排列,或围绕腺体漩涡状分布,周围扩张的隐窝常伴锯齿状结构。免疫表型:温和的梭形细胞不同程度表达EMA和CD34。随访4例患者均未见肿瘤复发和转移。结论胃肠道神经束膜瘤是一种良性的外周神经鞘膜肿瘤,可能被误诊为其他更常见的胃肠道梭形细胞肿瘤。免疫组化标记EMA和CD34阳性有助于鉴别诊断,患者预后良好。 相似文献
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目的探讨甲状腺癌(TC)发生发展的潜在生物学功能,筛选TC的关键基因和通路。方法从GEO数据库中下载3组基因芯片数据集,分析TC组织与正常组织差异表达基因(DEGs),利用多种在线分析软件对DEGs进行功能富集、通路分析、构建蛋白质互作网络、筛选关键基因,然后采用TCGA数据库对关键基因进行验证及生存分析。结果3组数据集分析得出410个共同DEGs,其中159个基因上调,251个基因下调。DEGs与癌症中的蛋白聚糖、P53信号通路、ECM-受体相互作用、细胞周期等信号通路有密切关系。筛选出14个关键基因,分别是CCNB2、FN1、MMP9、TIMP1、CXCL8、VCAN、EVA1A、LGALS1、KIF15、KIF20A、KIF4A、TOP2A、JUN和SDC2,其中MMP9、SDC2、KIF15和VCAN影响患者生存率,提示可以作为TC的潜在预后标志物。结论利用生物信息学方法筛选出14个关键基因和通路,可能有助于甲状腺癌的早期分子诊断与基因靶向治疗。 相似文献
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目的 探讨大鼠肝硬变和肝癌发生中肝组织病理和甲胎蛋白变化的意义.方法 选取雄性Wistar大鼠,分别采用DENA、四氧化碳和橄榄油诱导建立肝癌和肝硬变模型,于诱导前和诱导后2周、4周、8周、14周、18周和21周分别获取肝、脾组织和外周静脉血,HE染色进行肝脾组织病理学检查,ELISA法测定外周血清AFP水平.结果 大鼠肝硬变诱发过程中18周后出现肝假小叶,淤血性脾肿大;大鼠肝癌诱发过程中14周病理性核分裂,核仁变大、数量增多,肝癌结节形成,脾脏充血性改变.大鼠肝硬变诱发过程中外周血AFP在4周开始升高,14周与对照组比较差异显著(P<0.05);大鼠肝癌诱发过程中外周血AFP在2周开始升高,8周时与对照组比较差异显著(P<0.05);肝癌大鼠外周血AFP表达水平在8周时显著高于肝硬变大鼠(P<0.05).结论 不同诱导因素下大鼠肝脏病理变化出现时间和损害程度不尽相同,DENA对肝脏损害程度较四氯化碳对肝脏损害重;肝细胞受到病毒、细茵、毒素、化学毒物等损害时,动态监测外周血AFP水平,对评估肝细胞损伤程度和癌变具有双重生物学意义. 相似文献