TAT-凋亡素融合蛋白的表达及其抗肿瘤活性

凋亡素(apoptin)由鸡贫血病毒vp3基因编码,能特异地诱导肿瘤细胞凋亡而对正常细胞 没有毒性,为了获得可转导入细胞内部的凋亡素,将人工合成的编码TAT蛋白转导结构域的DNA片段与凋亡素编码基因克隆入质粒pET-28a内,构建出表达融合蛋白TAT-apoptin的原核表达载体pET-28a-TAT-apoptin.在大肠杆菌Rosetta（DE3）中表达融合蛋白,利用IDA -Ni2+ 亲和柱纯化,葡聚糖凝胶G 25除去尿素后得到可溶的变性蛋白.纯化后的TAT apoptin加入体外培养的人脐静脉内皮细胞(HUVECs)和人肺癌Anip973细胞,对照组加入TAT-麦芽糖结合蛋白（TAT-MBP）. 经免疫组化检测,转导1 h后TAT-MBP分布于以上两种细胞的胞浆和胞核,TAT-apoptin则主要分布于2种细胞的胞浆内,转导24 h后TAT-MBP的亚细胞定位没有变化,TAT-apoptin分别定位于HUVECs的胞浆和Anip973的胞核中.脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（TUNEL）显示转导48 h后,TAT-MBP处理过的 HUVECs和Anip973细胞、TAT-apoptin处理过的HUVECs没有明显改变,而TAT-apoptin处理过的Anip973细胞大量凋亡.以上结果表明TAT apoptin融合蛋白在肿瘤治疗上有潜在的应用价值.     

胶原样分子与肿瘤关系研究进展

细胞外基质不仅维持着体内细胞微环境的稳定,还在细胞的正常生长、增殖以及细胞之间的信号传导中起着重要作用。肿瘤发生时,基质中的分子组分发生了改变,这些改变朝着有利于肿瘤细胞生长侵袭的方向发展。在这个过程中,细胞外基质的主要成分在合成和分解上发生巨大变化,胶原分子便是其中之一,胶原分子作为细胞外基质中的主要成分,对细胞的黏附、运动、迁移等活动起着重要作用。随着研究的深入,发现越来越多的胶原分子参与了肿瘤的发生发展。基质中还存在着一些分子,它们在结构上和胶原蛋白一样含有三螺旋胶原结构域,在肿瘤的发生发展过程中同样发挥着重要作用。本文就包括胶原分子在内的含有胶原结构的分子在肿瘤中的作用做一综述。     

人卵巢浆液性囊腺癌永生化细胞系BUPH∶OVCA-3的建立及其生物学特性

介绍人卵巢浆液性囊腺癌永生化细胞系的建立 ,研究其生物学特性 .以卵巢浆液性乳头状囊腺癌的腹水细胞为材料 ,进行体外培养 .将永生化基因———SV4 0T抗原基因转染第 2代细胞 ,得到永生化细胞系 .通过光学显微镜、生长曲线测定、染色体分析、双层软琼脂培养、裸鼠接种、免疫组化等 ,研究其生物学特性 ,并与其来源细胞的生物学特性进行比较 .建立了一株人卵巢浆液性囊腺癌永生化细胞系 ,命名为BUPH∶OVCA 3,现已传至 6 0余代 .其生物学特性为 ,细胞生长旺盛 ;具有人体恶性细胞的核型特征 ;细胞恶性度较低 ,不具有集落形成能力及裸鼠接种致瘤性 ;除较未永生化细胞生长速率增快 ,饱和密度增加外 ,仍保留上皮细胞的分化表型 .结果表明 ,BUPH∶OVCA 3为一株恶性度较低的人卵巢浆液性囊腺癌永生化细胞系 ,保留其来源细胞的生物学特性 ,可作为研究恶性度较低的卵巢上皮癌的体外模型     

细胞周期抑制蛋白p27的研究进展

p27基因位于人类染色体12p13,其编码的蛋白对cyclinsCDKs具有广泛的抑制活性,是细胞周期调控的抑制蛋白。它以化学剂量的方式调节细胞周期的进程,参与细胞的生长、分化等过程。对p27基因的发现、基因结构、对细胞周期和细胞分化的调控机制以及与肿瘤的关系作一介绍。     

Mathematical modeling links Wnt signaling to emergent patterns of metabolism in colon cancer

Cell‐intrinsic metabolic reprogramming is a hallmark of cancer that provides anabolic support to cell proliferation. How reprogramming influences tumor heterogeneity or drug sensitivities is not well understood. Here, we report a self‐organizing spatial pattern of glycolysis in xenograft colon tumors where pyruvate dehydrogenase kinase (PDK1), a negative regulator of oxidative phosphorylation, is highly active in clusters of cells arranged in a spotted array. To understand this pattern, we developed a reaction–diffusion model that incorporates Wnt signaling, a pathway known to upregulate PDK1 and Warburg metabolism. Partial interference with Wnt alters the size and intensity of the spotted pattern in tumors and in the model. The model predicts that Wnt inhibition should trigger an increase in proteins that enhance the range of Wnt ligand diffusion. Not only was this prediction validated in xenograft tumors but similar patterns also emerge in radiochemotherapy‐treated colorectal cancer. The model also predicts that inhibitors that target glycolysis or Wnt signaling in combination should synergize and be more effective than each treatment individually. We validated this prediction in 3D colon tumor spheroids.     

胃肠道间质瘤组织中MMP-13与CD147的表达及其意义

目的探讨基质金属蛋白酶-13（Matrix metalloproteinases-13,MMP-13）和CD147的表达与胃肠道间质瘤（Gastrointestinals stromal tumors,GISTs）临床病理特征的关系。方法采用免疫组织化学检测69例GISTs组织中MMP-13及CD147的表达,分析MMP-13和CD147与各病理参数的关系。结果GISTs组织中MMP-13和CD147的表达率分别为95.7%和97.1%,MMP-13阳性表达程度与GISTs的生物学行为、CD147表达及肿瘤的大小呈正相关（P〈0.05）;CD147阳性表达程度与肿瘤的大小呈正相关（P〈0.05）。结论MMP-13表达与GISTs生物学行为关系密切,可作为GISTs生物学行为的潜在评价指标。     

基于碳酸酐酶IX的肿瘤成像和治疗研究进展

碳酸酐酶IX (carbonic anhydrase IX, CAIX)是一种在乏氧肿瘤细胞表面特异性过表达的跨膜蛋白,具有调节肿瘤细胞内外酸碱度的功能,与肿瘤增殖、侵袭和转移息息相关。因此,CAIX是一个很有潜力的肿瘤成像和治疗靶点。本文详细阐述了基于CAIX的肿瘤成像、治疗和诊疗一体化的研究进展,并对CAIX作为抗肿瘤靶点的应用前景进行了展望。     

lncRNA与miRNA相互调控作用及其与肿瘤的关系研究

长链非编码RNA (long non-coding RNA,lncRNA)是一类转录本长度大于200 bp的非编码RNA,可作为人类基因组中一类重要的调控分子通过多种方式发挥其生物学功能.近年来的研究表明,lncRNA也可以作为一种竞争性内源性RNA (competing endogenous RNA, ceRNA) 与miRNA相互作用,参与靶基因的表达调控,并在肿瘤的发生发展中发挥重要的作用.本综述在简要介绍lncRNA功能研究现状和主要研究方法的基础上,进一步分析了lncRNA与miRNA之间的互相调控关系及其在肿瘤发生发展中的作用,以便为后续的研究提供新的思路.     

改良法建立妊娠期高血压疾病大鼠模型

目的寻找理想的妊娠期高血压疾病动物模型。方法60只雌性Wistar大鼠分为六组,分别为正常妊娠组（A组）、左旋亚硝酸精氨酸甲酯（L-NAME）腹腔注射组（B组）、生理盐水注射组（C组）、假手术组（D组）、双侧子宫动脉结扎术组（E组）以及L-NAME腹腔注射联合腹主动脉缩窄术组（F组）,对大鼠血压、尿蛋白值以及胎鼠、胎盘、胎鼠头的重量进行监测并观察肾脏、胎盘的病理改变。结果F组孕鼠于孕13d即出现了蛋白尿、血压升高,较B组及E组出现时间早（P〈0.05）,其胎鼠体重、胎头重量下降较各组更明显（P〈0.05）,肾小球小动脉管壁增厚、管腔狭窄;胎盘出现血管间膜增厚、纤维蛋白沉积等妊娠高血压疾病的典型病理改变。结论L-NAME腹腔注射联合腹主动脉缩窄术是更为理想的建立妊娠高血压疾病动物模型的方法。     

Three-dimensional in vitro tumor models for cancer research and drug evaluation

Cancer occurs when cells acquire genomic instability and inflammation, produce abnormal levels of epigenetic factors/proteins and tumor suppressors, reprogram the energy metabolism and evade immune destruction, leading to the disruption of cell cycle/normal growth. An early event in carcinogenesis is loss of polarity and detachment from the natural basement membrane, allowing cells to form distinct three-dimensional (3D) structures that interact with each other and with the surrounding microenvironment. Although valuable information has been accumulated from traditional in vitro studies in which cells are grown on flat and hard plastic surfaces (2D culture), this culture condition does not reflect the essential features of tumor tissues. Further, fundamental understanding of cancer metastasis cannot be obtained readily from 2D studies because they lack the complex and dynamic cell–cell communications and cell–matrix interactions that occur during cancer metastasis. These shortcomings, along with lack of spatial depth and cell connectivity, limit the applicability of 2D cultures to accurate testing of pharmacologically active compounds, free or sequestered in nanoparticles. To recapitulate features of native tumor microenvironments, various biomimetic 3D tumor models have been developed to incorporate cancer and stromal cells, relevant matrix components, and biochemical and biophysical cues, into one spatially and temporally integrated system. In this article, we review recent advances in creating 3D tumor models employing tissue engineering principles. We then evaluate the utilities of these novel models for the testing of anticancer drugs and their delivery systems. We highlight the profound differences in responses from 3D in vitro tumors and conventional monolayer cultures. Overall, strategic integration of biological principles and engineering approaches will both improve understanding of tumor progression and invasion and support discovery of more personalized first line treatments for cancer patients.