YAP1基因促进人甲状腺癌细胞生长与迁移

目的 构建带myc标签的人Yes相关转录调节蛋白1(YAP1)基因的真核表达载体,探讨YAP1对甲状腺癌BCPAP细胞增殖和迁移的影响.方法 采用聚合酶链反应(PCR)扩增YAP1的编码区序列,将其插入到pXJ40-myc载体中,构建pXJ40-myc-YAP1重组质粒并鉴定;质粒转染人甲状腺癌BCPAP细胞,Western印迹鉴定融合蛋白表达,细胞增殖实验(CCK-8法)、克隆形成实验、划痕实验检测细胞的生长、增殖和迁移能力的变化.结果 pXJ40-myc-YAP1质粒构建成功;转染BCPAP细胞后重组融合蛋白表达成功;细胞增殖实验、克隆形成及划痕实验证明YAP1可促进甲状腺癌细胞BCPAP的增殖和迁移.结论 带myc标签的人YAP1基因能在人甲状腺癌BCPAP细胞中表达,且可促进该细胞的增殖和迁移,为进一步研究YAP1在甲状腺肿瘤中的功能奠定了基础.     

溶血磷脂酸酰基转移酶β促进人肺成纤维细胞的增殖、迁移与黏附

目的分析溶血磷脂酸酰基转移酶p（1ysophosphatidic acid acyltransferase,LPAATp）在多种细胞系中的表达情况及其过表达对细胞体外增殖、迁移与黏附的影响。方法通过免疫印迹实验检测LPAATp在多种细胞中的表达水平;构建pcDNA3．1A（＋）／LPAATB真核表达载体,转染人肺成纤维细胞（humanlung fibroblast,HLF）并筛选稳定转染细胞株;通过MTY细胞增殖实验分析过表达LPAATB对HLF细胞体外增殖的作用,通过划痕修复实验分析稳定株细胞的体外迁移能力,利用细胞黏附实验分析稳定株细胞对细胞外基质成分Matrigel的黏附情况。结果LPAAT8蛋白在肿瘤细胞中表达水平存在差异。体外生物学实验显示,稳定转染LPAATB的HLF细胞增殖能力明显增强,细胞迁移能力显著提高,与细胞外基质成分Matrigel的黏附能力显著增强。结论LPAAT家族成员LPAAT8在人类多种细胞中的表达具有普遍性,其过表达明显促进HLF细胞的增殖、迁移及黏附能力,提示该基因在细胞体外生长过程中具有重要作用。     

抑制CHD6表达对A549细胞辐射敏感性的影响

目的通过RNA干扰技术建立染色质重构蛋白CHD6基因表达抑制的细胞模型,研究CHD6对人肺腺癌细胞A549细胞增殖及辐射敏感性的影响.方法利用质粒介导的siRNA技术建立CHD6基因表达抑制细胞模型,RT-PCR检测CHD6 mRNA的表达,细胞生长曲线和流式细胞技术分别检测A549细胞增殖及细胞周期的变化,荧光染色法检测细胞凋亡,细胞克隆形成率检测A549细胞辐射敏感性.结果通过本研究,成功构建了siRNA抑制CHD6表达的细胞模型;通过siRNA抑制CHD6的表达,A549细胞增殖能力明显增强,细胞对2 Gy以内γ射线照射有明显辐射抗性;大剂量照射后的细胞凋亡率无明显改变.结论抑制CHD6基因表达将提高细胞增殖能力和细胞的辐射抗性.     

蛋白酶体抑制剂MG132联合X射线对人肺腺癌细胞生长迁移和周期的影响及作用机制

目的 研究蛋白酶体抑制剂MG132联合X射线对人肺腺癌A549细胞生长、迁移、侵袭、细胞周期分布的影响及机制。方法 MTT法检测不同MG132浓度处理后不同时间肺腺癌A549细胞的增殖;克隆形成实验检测A549细胞的存活能力;划痕实验测定A549细胞的迁移能力;Transwell小室测定其侵袭能力;流式细胞术测定细胞周期分布的改变;Western blot法测定蛋白表达水平。结果 MG132明显抑制人肺腺癌A549细胞的生长,并呈现剂量-效应和时间-效应关系。MG132联合X射线对A549细胞克隆存活能力有显著抑制作用（F=554.78、954.64,P<0.01）,且加MG132组克隆存活率均低于照射组（t=4.44、12.41、3.52、6.72,P<0.05）。MG132在无毒性剂量下联合X射线可显著抑制A549细胞的迁移及侵袭能力（t=12.79,P<0.01）,并明显增加G₁期细胞阻滞（t=4.29,P<0.05）;MG132联合X射线明显降低A549细胞中基质金属蛋白酶-2,-9和G₁期相关蛋白Cyclin D1的表达水平,同时增加细胞中P53的表达。结论 MG132可以显著抑制人肺腺癌A549细胞的生长,且在无毒性剂量下联合X射线可以明显降低其转移和侵袭能力,显著增加肿瘤细胞的G₁期阻滞。     

SC-58236与顺铂联合调控人肺腺癌A549细胞增殖及肺癌移植瘤生长的作用

目的：观察高选择性COX-2抑制剂SC-58236和顺铂联合对人肺腺癌A549细胞增殖及移植瘤生长的影响。方法：采用MTT法观察A549细胞增殖,同时用PTENsiRNA转染A549细胞,并将其接种裸鼠,观察SC-58236和／或顺铂对A549细胞增殖及移植瘤生长的影响。结果：SC-58236可增强顺铂对A549细胞增殖及其对移植瘤生长的抑制作用,并协同促进细胞及瘤组织内PTEN蛋白表达（P〈0．01）。PTENsiRNA转染可减弱顺铂与SC-58236对A549细胞增殖及肿瘤生长的抑制作用（P〈0．01）。结论：SC-58236可增强顺铂对A549细胞的细胞毒性,该作用与增加PTEN蛋白表达有关。     

Effect of HIP1 gene silencing on proliferation of PC-3 cells in human androgen-independent prostate tumor

目的 研究HIP1基因沉默对人前列腺癌PC-3细胞增殖的影响.方法 构建靶向HIP1的shRNA表达载体pSilence-shHIP1,应用脂质体转染到PC-3细胞,RT-PCR检测HIP1基因沉默效果,Western blotting确认有效作用靶点.通过细胞划痕实验和细胞生长曲线研究HIP1基因对PC-3细胞增殖的影响.结果 转染PC-3细胞后,沉默HIP1基因效率达83％(P＜0.01); Western blotting证实该基因片段为抑制HIP1的有效作用靶点.细胞划痕实验和生长曲线显示HIP1基因沉默可抑制PC-3细胞的增殖.结论 pSilence-shHIP1表达载体可特异性地抑制HIP1基因的表达,沉默HIP1基因可抑制PC-3细胞增殖和迁移.     

S100A4 siRNA对人胰腺癌BxPc-3细胞增殖、迁移及侵袭的影响

目的观察利用小干扰RNA（small interference RNA,siRNA）技术下调人胰腺癌BxPc-3细胞中钙结合蛋白S100A4表达后对细胞增殖、迁移和侵袭的影响。方法设计合成针对S100A4的特异性siRNA,转染至人胰腺癌BxPc-3细胞,以Western印迹检测转染前后S100A4蛋白表达变化;克隆形成率实验检测转染前后BxPc-3细胞增殖能力;Transwell小室模型检测转染前后BxPc-3细胞迁移和侵袭能力变化。结果 Western印迹结果显示BxPc-3细胞转染S100A4siRNA48h后,S100A4蛋白表达明显下调;Transwell小室验证转染S100A4siRNA后细胞的迁移和侵袭能力明显降低。结论本研究结果表明,S100A4表达下调后可抑制胰腺癌细胞的增殖、迁移和侵袭,提示S100A4可以作为一个潜在的肿瘤治疗靶标。     

稳定低表达CD147的人肺腺癌细胞系的建立及对细胞增殖的影响

目的构建CD147慢病毒干涉载体,建立稳定下调CD147表达的人肺腺癌A549细胞系,观察下调CD147表达后人肺腺癌细胞增殖能力的变化。方法将携带不同特异性干涉序列的3个DNA片段分别插入干涉质粒pSicoR中,构建pSicoR-siCD147慢病毒干涉载体(pSicoR-siCD147-1,pSicoR-siCD147-2,pSicoR-siCD147-3)。分别将构建好的载体转染入293FT细胞中,进行慢病毒包装。建立稳定下调CD147表达的细胞系。利用CCK-8法检测细胞增殖能力的变化。结果与结论构建了pSicoR-siCD147慢病毒干涉载体,RT-PCR和实时PCR结果显示pSicoR-siCD147-2的干涉效率最高。建立了稳定下调CD147表达的人肺腺癌A549细胞系(A549-siCD147)。CCK-8法检测发现A549-siCD147细胞的增殖能力显著性下降(P<0.05),下调CD147的表达可以使人肺腺癌细胞的增殖能力受到明显抑制。     

靶向生存素基因的RNAi对肺腺癌A549细胞放射敏感性的影响

目的 构建靶向生存素(survivin)基因的RNA干扰(RNAi)载体,观察对肺腺癌A549细胞放射敏感性的影响,并探讨其机制.方法 根据survivin的cDNA序列设计干扰序列,构建干扰survivin的重组干扰质粒pGenesil2-survivin.酶切、测序鉴定正确后,经脂质体介导转染肺腺癌A549细胞;应用反转录.聚合酶链式反应(RT-PCR)和Western blot法检测survivin的表达;流式细胞术检测细胞凋亡的变化;克隆形成实验检测细胞的放射敏感性.结果 酶切和测序结果显示,载体构建正确;将质粒pGenesil2-survivin转染肺腺癌A549细胞48 h,survivin蛋白水平及mRNA水平在正常组与pGenesil2组中无明显变化,5 Gy照射组增强,而转染pGenesil2-survivin后明显抑制;pGenesil2-survivin与5 Gy X射线照射都能诱导细胞凋亡增加(t1=10.63,P<0.001;t2=3.75,P<0.05),两者共同作用,凋亡增加更明显(t=4.83,P<0.05);克隆形成实验显示,pGenesil2与正常组D0、Dq无明显变化,而pGenesil2.survivin则明显降低,说明其可以提高A549细胞的放射敏感性.结论 靶向生存素基因的RNAi能够明显抑制survivin mRNA与蛋白的表达,促进凋亡,增强A549细胞的放射敏感性.     

18F-Fluoroestradiol

Estrogen receptor (ER) expression is an important determinant of breast cancer behavior and is critical for response to endocrine therapies such as tamoxifen and aromatase inhibitors. In current practice, ER expression is determined by assay of biopsy material. In more advanced disease, tissue assay may present practical difficulties and be associated with significant sampling error. This and other considerations motivated the development of ER imaging agents for positron emission tomography (PET), of which the most successful has been (18)F-16alpha-17beta-fluoroestradiol (FES). In this review, we highlight aspects of ER biology and the importance of the ER in breast cancer therapy; review the structure and synthesis of FES; describe its kinetics and safety/dosimetry data; and highlight validation studies. Also discussed are early results in patients using FES-PET to localize ER-expressing tumors and associated data pointing toward its accuracy as a predictive assay for breast cancer endocrine therapy. Finally, early data for tumors and sites other than breast cancer are mentioned. Preliminary data strongly point toward potential clinical utility for FES-PET, motivating further validation and future clinical trials with prospective endpoints tested under appropriate regulatory oversight.     

Comparative uptake of 18F-FEN-DPAZn2, 18F-FECH, 18F-fluoride,and 18F-FDG in fibrosarcoma and aseptic inflammation

The aim of this study is to evaluate uptake of 2-¹⁸F-fluoroethyl-bis(zinc(II)-dipicolylamine) (¹⁸F-FEN-DPAZn2) as a promising cell death imaging agent, a choline analog ¹⁸F-fluoroethylcholine (¹⁸F-FECH), ¹⁸F-fluoride as a bone imaging agent, and a glucose analog 2−¹⁸F-fluoro-2-deoxy-d-glucose (¹⁸F-FDG) in the combined S180 fibrosarcoma and turpentine-induced inflammation mice models. The results showed that ¹⁸F-FDG had the highest tumor-to-blood uptake ratio and tumor-to-muscle ratio, and high inflammation-to-blood ratio and inflammation-to-muscle ratio. ¹⁸F –FECH showed moderate tumor-to-blood ratio and tumor-to-muscle ratio, and low inflammation-to-blood ratio and inflammation-to-muscle ratio. However, accumulation of ¹⁸F FEN-DPAZn2 in tumor was similar to that in normal muscle. Also, ¹⁸F-FEN-DPAZn2 and ¹⁸F-fluoride exhibited the best selectivity to inflammation. ¹⁸F-FECH positron emission tomography (PET) imaging demonstrates some advantages over ¹⁸F-FDG PET for the differentiation of tumor from inflammation. ¹⁸F FEN-DPAZn2 and ¹⁸F-fluoride can be used for PET imaging of aseptic inflammation.     

A simple 18O water target for 18F production

A simple low-volume (3 mL) target has been constructed for the production of 18F via the proton irradiation of 18O enriched water. The target is constructed of copper with the water cavity electroplated with nickel. This target routinely produces more than a curie of 18F (as [18F]fluoride) suitable for radiopharmaceutical syntheses.     

18例创伤性巨大膈疝的诊治

目的：总结创伤性巨大膈疝发病特点，提高膈疝早期诊断率，降低死亡率．方法：通过回顾性总结我院１０年来创伤性巨大膈疝的诊治经过，以实例分析对比的方法，总结出创伤性巨大膈疝发病特征、诊治规律及误诊原因．结果：本组１８例创伤性巨大膈疝，全部给予开胸探查、脏器复位后行膈肌修补术，１６例治愈；２例死亡．结论：创伤性巨大膈疝早期诊断、立即手术治疗，是提高治愈率、降低死亡率的关键     

Studies on 18F-labeled pyrimidines. Tumor uptakes of 18F-5-fluorouracil, 18F-5-fluorouridine, and 18F-5-fluorodeoxyuridine in animals

Three 18F-labeled pyrimidines, 18F-5-fluorouridine (18F-5-FUR), 18F-5-fluorouracil (18F-5-FU), and 18F-5-fluorodeoxyuridine (18F-5-FdUR), were examined regarding tissue distribution and tumor uptake in ascitic hepatoma AH109A-bearing rats. The differential absorption ratios of tumors of 18F-5-FUR, 18F-5-FU, and 18F-5-FdUR were 0.75 +/- 0.21, 0.92 +/- 0.15, and 0.96 +/- 0.24 at 30 min, and 0.37 +/- 0.09, 0.64 +/- 0.34, and 0.60 +/- 0.17 at 120 min, respectively. The tumor-to-organ ratios obtained with three radiopharmaceuticals, especially with blood, heart, lung, muscle, and brain were high and these ratios increased with time. The tumor-to-organ ratios obtained with 18F-5-FdUR were always 1.3-4 times higher than 18F-5-FU and 18F-5-FUR. We concluded that 18F-5-FdUR was a suitable radiopharmaceutical for tumor imaging. Positron emission tomography of a rabbit tumor located on the chest with 18F-5-FdUR clearly showed the tumor within 1 h.