伊马替尼联合P27基因克隆对慢性粒细胞白血病K562细胞的作用

目的:探讨伊马替尼和P27基因克隆联合作用对K562细胞的细胞增殖、细胞周期及凋亡等的调控作用。方法:RT-PCR扩增P27基因,胶纯化回收后连接到T载体测序,序列正确后构建P27-pcDNA3.1载体,将P27-pcDNA3.1与空载体分别以脂质体转染入P27基因缺失的K562细胞,经筛选得到G418抗性的K562细胞株,Westernblot证实转染后有P27蛋白表达,MTT法检测细胞生长指数和细胞存活率,流式细胞仪检测细胞周期和凋亡。结果:表达外源性P27蛋白的P27-pcDNA3.1-K562细胞株生长速度明显慢于对照K562细胞株,流式细胞仪显示G0/G1期细胞增多,S期细胞明显减少,P27-pcDNA3.1-K562细胞与伊马替尼联合应用后凋亡细胞比例明显上升,MTT法显示细胞存活率较P27-K562细胞组及伊马替尼-K562细胞组均明显下降(P<0.01vsP27-K562细胞组,P<0.05vs伊马替尼-K562细胞组)。结论:表达外源P27蛋白联合应用伊马替尼对抑制K562细胞增殖及促凋亡方面具有协同作用。     

乙肝相关性肝癌组织中p27、p53及增殖细胞核抗原的表达

目的探讨乙肝相关性肝细胞癌组织中p27、p53与增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）的表达及其临床病理学意义。方法通过免疫组化法检测40例乙肝相关性肝细胞肝癌组织及其相应的癌旁组织中p27、p53与PCNA的表达。结果（1）p53蛋白在肝癌组织中阳性表达率明显高于癌旁肝组织（P=0.047），低分化肝癌组织中p53阳性表达率明显高于高中分化者（P=0．017）。（2）肝癌组织中p27蛋白染色指数明显低于癌旁肝组织（P=0．045），在肿瘤直径〉5cm（P=0．031）与存在肝外转移的肝癌组织中进一步降低（P=0．028）。（3）肝癌组织中PCNA染色指数明显高于癌旁肝组织（P=0.029）。在肿瘤直径〉5cm（P=0．037）与包膜侵犯的肝癌组织中进一步增高（P=0．035）。（4）Cox比例风险模型多因素分析发现临床分期（P=0．028）为独立的预后因子。结论细胞周期调控因子p53、p27与PCNA的异常表达与乙肝相关性肝细胞癌的分化、浸润及转移有着密切的联系。     

甲状腺癌组织中p27和增殖细胞核抗原的表达及其相关性分析

目的探讨甲状腺癌组织中p27和增殖细胞核抗原(PCNA)的表达情况及其相关性。方法用免疫组织化学法检测86例甲状腺癌(甲状腺癌组)、54例甲状腺腺瘤(腺瘤组)和瘤旁正常甲状腺组织(正常组)中p27和PCNA的表达情况。结果 p27在甲状腺癌组阳性表达率分别为37.21%,高于腺瘤组和正常组,差异有统计学意义(P<0.01)。PCNA在甲状腺癌组阳性表达率为66.27%,高于腺瘤组和正常组,差异有统计学意义(χ2=22.16,P<0.01)。p27在甲状腺乳头状癌、滤泡癌、髓样癌和未分化癌中的阳性表达率分别为48.94%、31.25%、14.29%和11.11%,差异有统计学意义(χ2=8.85,P<0.05);PCNA在乳头状癌、滤泡癌、髓样癌和未分化癌中的阳性表达率分别为53.19%、75%、85.71%和88.89%,差异有统计学意义(χ2=8.57,P<0.05)。甲状腺癌p27和PCNA表达呈负相关(r=-0.51,P<0.01)。结论 p27和PCNA在甲状腺癌中的阳性表达明显减少,并且随着分化程度降低减少更为明显;而PCNA在甲状腺癌中的阳性表达明显增加,并且随分化程度降低增加更为明显;并且p27和PCNA表达呈负相关。     

转化生长因子β1和即刻早期基因及p27在中耳胆脂瘤上皮中的表达

目的　研究转化生长因子 β1(transforminggrowthfactor β1,TGF β1)、即刻早期基因 (c fos)、周期素依赖性激酶抑制蛋白 p2 7在中耳胆脂瘤上皮中的表达及相互之间的关系 ,探讨其表达对胆脂瘤侵袭能力的影响。方法　应用免疫组化链霉菌抗生物素蛋白 过氧化酶染色法检测TGF β1、c fos和 p2 7在 31例胆脂瘤上皮、11例胆脂瘤患者外耳道上皮和 10例正常外耳道上皮中的表达 ,应用计算机图像分析系统对其阳性表达进行定量分析。结果　TGF β1和c fos在胆脂瘤上皮中表达阳性率分别为 87 1%和 83 9% ,与胆脂瘤患者外耳道上皮和正常外耳道上皮相比表达差异有显著性。而p2 7在胆脂瘤上皮和外耳道上皮组上皮中均未见阳性表达。胆脂瘤上皮中c fos与TGF β1表达无相关性 (r =0 339,P =0 331) ;胆脂瘤侵袭能力与c fos表达有高度显著相关关系 (r =- 0 96 5 ,P <0 0 1) ,与TGF β1表达也有高度相关关系 (r =- 0 4 0 6 ,P <0 0 1)。 结论　即刻早期基因c fos在胆脂瘤中表达显著增强 ,且与胆脂瘤的侵袭能力有高度显著相关性 ,提示高c fos表达是胆脂瘤高增殖特征的因素之一。TGF β1在胆脂瘤上皮中高表达亦表明其在胆脂瘤的高增殖能力方面起一定作用     

Elucidation of the Mechanism of Host NMD Suppression by HTLV-1 Rex: Dissection of Rex to Identify the NMD Inhibitory Domain

The human retrovirus human T-cell leukemia virus type I (HTLV-1) infects human T cells by vertical transmission from mother to child through breast milk or horizontal transmission through blood transfusion or sexual contact. Approximately 5% of infected individuals develop adult T-cell leukemia/lymphoma (ATL) with a poor prognosis, while 95% of infected individuals remain asymptomatic for the rest of their lives, during which time the infected cells maintain a stable immortalized latent state in the body. It is not known why such a long latent state is maintained. We hypothesize that the role of functional proteins of HTLV-1 during early infection influences the phenotype of infected cells in latency. In eukaryotic cells, a mRNA quality control mechanism called nonsense-mediated mRNA decay (NMD) functions not only to eliminate abnormal mRNAs with nonsense codons but also to target virus-derived RNAs. We have reported that HTLV-1 genomic RNA is a potential target of NMD, and that Rex suppresses NMD and stabilizes viral RNA against it. In this study, we aimed to elucidate the molecular mechanism of NMD suppression by Rex using various Rex mutant proteins. We found that region X (aa20–57) of Rex, the function of which has not been clarified, is required for NMD repression. We showed that Rex binds to Upf1, which is the host key regulator to detect abnormal mRNA and initiate NMD, through this region. Rex also interacts with SMG5 and SMG7, which play essential roles for the completion of the NMD pathway. Moreover, Rex selectively binds to Upf3B, which is involved in the normal NMD complex, and replaces it with a less active form, Upf3A, to reduce NMD activity. These results revealed that Rex invades the NMD cascade from its initiation to completion and suppresses host NMD activity to protect the viral genomic mRNA.     

地塞米松抑制人卵巢癌细胞HO-8910增殖的分子机制:RhoB信号通路的作用

目的 观察地塞米松（Dex）对人卵巢癌细胞HO-8910增殖的影响,探讨小G蛋白RhoB信号通路在其中的作用.方法四甲基偶氮唑盐比色法（MTT法）和软琼脂实验检测细胞的增殖;RT-PCR法检测RhoB mRNA的表达;Western印迹检测RhoB及其上下游信号分子磷酸化Akt（p-Akt）、p21^cip1/waf1和p27蛋白质的表达;报告基因技术检测p21^cip1/waf1基因的转录.结果Dex可以明显抑制HO-8910细胞锚依赖性和非依赖性方式的增殖,100 nM Dex处理细胞6 d,抑制率为31%.100nM Dex处理细胞可以明显上调RhoB mRNA和蛋白质的表达（为对照的2.5倍）;同时伴有p-Akt蛋白质表达的降低（24 h时约为对照的50%）、p27蛋白质表达的增加（36 h时比对照增加了3.3倍）以及p21^cip1/waf1转录和蛋白质表达增多（24h时约为对照的1.7倍）.RhoB过表达可以降低细胞的生长速度,并能增强Dex的增殖抑制作用（100 nM Dex处理6 d,抑制率为44%）;而RhoB表达阻断后可逆转Dex的这一效应（抑制率约为13%）.结论Dex抑制HO-8910细胞增殖的机制可能与抑制PI3K/p-Akt信号,从而上调RhoB蛋白质,使RhoB下游信号分子p21^cip1/waf1和p27蛋白质表达增加有关.     

p27蛋白、细胞周期素E和增殖细胞核抗原在上皮性卵巢肿瘤的表达及其临床意义

目的探讨p27、细胞周期素E(CyclinE)和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在上皮性卵巢肿瘤组织中的表达及其与上皮性卵巢癌的临床分期、分化程度和病理类型的关系。方法应用流式细胞技术检测64例上皮性卵巢肿瘤组织中p27、CyclinE和PCNA的表达,并进行对比分析。结果p27在卵巢癌组织中呈低表达(P<0.01),而CyclinE和PCNA在卵巢癌组织中呈高表达(P<0.01,P<0.05);在Ⅲ~Ⅳ期和低分化患者中p27表达下降,而CyclinE和PCNA表达升高。p27与CyclinE和PCNA表达呈负相关(r=-0.532,P<0.01,r=-0.564,P<0.01),CyclinE和PCNA表达呈正相关(r=0.556,P<0.05)。结论p27、CyclinE和PCNA的表达与上皮性卵巢肿瘤的临床分期和病理分级有关,而与病理类型无关。     

p27和p-ERK1/2在肝细胞癌中的表达及相关性研究

目的 探讨p27和p-ERK1/2在肝细胞癌(hepatocellular carcinoma,HCC)中的表达、意义及二者间可能的关系.方法 免疫组织化学(S-P)法检测肝癌50例、肝硬化15例及正常肝组织10例中p27和p-ERK1/2的表达.结果 肝癌组织中p27的阳性表达率(42%)明显低于肝硬化(78%)和正常肝组织(90%),表达水平与肿瘤恶性程度、大小、是否广泛转移等有关(P＜0.05);肝癌组织中p-ERK1/2的阳性表达率(36%)显著高于肝硬化(21.5%)和正常肝组织(11.6%),表达与恶性程度、转移等有关;肝癌组织中p27的表达与p-ERK1/2表达呈负相关(rs=-0.4812,P＜0.01).结论 p-ERK1/2基因的激活可以影响p27的表达,二者与肝癌的发生、发展有关.     

Richter’s syndrome of T-cell lineage (T rex lymphoma)

Abstract

By definition, Richter’s syndrome represents the transformation of low-grade B-cell lymphoma into high-grade B-cell lymphoma, usually refractory to treatment. Exceptional cases of transformation into very aggressive mature T-cell lymphomas have been described as an unusual manifestation of the syndrome in patients died after few months from the diagnosis, despite chemotherapy. The time is ripe to regroup these T lymphomas under a new pathological subset, through the unequivocal alternate naming of ‘T rex lymphoma’, by analogy with the aggressive behavior of the famous dinosaur (T. rex). In practice, it represents the transformation of low-grade B-cell lymphoma into high-grade T-cell lymphoma, burdened by a very poor prognosis, because of the underlying B-cell lymphoma, which negatively interferes with the immune response of the patient. Against this distinct lymphomatous T clone, the major therapeutic efforts should be addressed.     

Neurosensory and Sinus Evolution as Tyrannosauroid Dinosaurs Developed Giant Size: Insight from the Endocranial Anatomy of Bistahieversor sealeyi

Tyrannosaurus rex and other tyrannosaurid dinosaurs were apex predators during the latest Cretaceous, which combined giant size and advanced neurosensory systems. Computed tomography (CT) data have shown that tyrannosaurids had a trademark system of a large brain, large olfactory bulbs, elongate cochlear ducts, and expansive endocranial sinuses surrounding the brain and sense organs. Older, smaller tyrannosauroid relatives of tyrannosaurids developed some, but not all, of these features, raising the hypothesis that tyrannosaurid-style brains evolved before the enlarged tyrannosaurid-style sinuses, which might have developed only with large body size. This has been difficult to test, however, because little is known about the brains and sinuses of the first large-bodied tyrannosauroids, which evolved prior to Tyrannosauridae. We here present the first CT data for one of these species, Bistahieversor sealeyi from New Mexico. Bistahieversor had a nearly identical brain and sinus system as tyrannosaurids like Tyrannosaurus, including a large brain, large olfactory bulbs, reduced cerebral hemispheres, and optic lobes, a small tab-like flocculus, long and straight cochlear ducts, and voluminous sinuses that include a supraocciptal recess, subcondyar sinus, and an anterior tympanic recess that exits the braincase via a prootic fossa. When characters are plotted onto tyrannosauroid phylogeny, there is a two-stage sequence in which features of the tyrannosaurid-style brain evolved first (in smaller, nontyrannosaurid species like Timurlengia), followed by features of the tyrannosaurid-style sinuses (in the first large-bodied nontyrannosaurid tyrannosauroids like Bistahieversor). This suggests that the signature tyrannosaurid sinus system evolved in concert with large size, whereas the brain did not. Anat Rec, 303:1043–1059, 2020. © 2020 American Association for Anatomy