HINT2在甲状腺乳头状癌中的表达和功能研究

HINT2与多种肿瘤发生发展密切相关,在甲状腺癌中的功能还不清楚.该文使用组织芯片检测甲状腺乳头状癌组织中HINT2的表达,发现12例癌组织中HINT2蛋白表达水平显著高于正常组织(86％,P＜0.05).该文构建高表达和低表达HINT2的甲状腺乳头状癌细胞系K1稳定克隆后,使用CCK8和克隆形成实验检测HINT2对细...     

HPV16L1抗原表达肿瘤模型的建立

应用基因重组技术,构建pcDNA-L1真核表达载体,经限制性内切酶和序列分析,用脂质体转染技术将其转入B16细胞,G418稳定筛选后IFA法检测其表达,RT-PCR法检测HPV16L1 mRNA 的生成,并将转染细胞接种小鼠皮下,观察成瘤情况及RT-PCR法检测HPv16L1 mRNA的生成。结果,酶切鉴定证实重组质粒中插入的目的基因片段及载体大小、方向和插入位点均正确,在转染的 B16细胞中可见绿色荧光并检测到HPV16L1 mRNA的生成,接种的转染细胞在小鼠皮下可成瘤, 接种后1月在肿瘤组织中可检测到HPV16L1 mRNA的生成,B16细胞转染L1后其致瘤性与转染空载体组和野生型B16细胞组无明显差异。     

COX-2和IL-1在甲状腺乳头状癌组织中的表达

目的探讨COX-2和IL-1在甲状腺乳头状癌组织中的表达及其在肿瘤的发生和发展中的作用。方法收集武汉大学人民医院和武汉大学中南医院病理科2000-2006年手术切除及活检的甲状腺乳头状癌标本共40例,另取癌周围组织5例作对照。采用免疫组织化学方法观察各组组织内COX-2和IL-1的表达。利用HPIAS-2000图像分析系统测定COX-2和IL-1在癌及癌旁组中表达的平均光密度和平均阳性面积率。结果甲状腺乳头状癌组织中COX-2和IL-1呈高表达;癌旁组织中COX-2和IL-1呈低表达。图像分析结果显示两组间差异有显著性意义（P〈0.01）。结论IL-1可能通过诱导COX-2的表达,在促进肿瘤的发生和发展中起作用。     

人乳头瘤病毒16型L1和L2晚期蛋白在原核细胞中的表达

目的　HPV的某些型别尤其是HPV16 ,18型感染与宫颈癌的发生有密切的关系。宫颈癌在全球妇女癌症死亡原因中居第二位。在发展中国家 ,宫颈癌在妇女癌症死因中列第一位。尽管早期宫颈癌患者放疗及手术治疗效果较好 ,但对中晚期宫颈癌患者治疗效果却很差。因而 ,采取有效的措施预防HPV感染对于宫颈癌防治工作无疑是非常必要的。在对乳头瘤病毒结构和功能的研究中发现 ,以L1和L2为保护性抗原构建的疫苗株很可能在预防HPV感染及其引起的恶性肿瘤方面发挥重要作用。目前国际上常通过原核和真核表达系统获得L1和L2蛋白。本实验拟通过原核表…     

RASSF1A和SKP2在胃癌中的表达及与幽门螺杆菌L型感染关系的研究

目的研究幽门螺杆菌L型(Helicobacter pyloriL-form,H.pylori-L型)感染,Ras相关区域家族1A基因(RASSF1A)和细胞S期激酶相关蛋白2(Skp2)在胃癌发生、发展中的作用及相互关系。方法应用革兰染色和免疫组织化学SP法检测50例胃癌组织及20例癌旁正常组织中的H.pylori-L型感染情况;同时应用逆转录聚合酶链式反应(RT-PCR)和免疫组织化学SP法检测上述组织中癌基因skp2和抑癌基因RASSF1A的表达。结果 50例胃癌组织标本中免疫组化及革兰染色H.pylori-L型同时阳性的病例有30例,胃癌组织和癌旁正常组织中H.pylo-ri-L型阳性率分别为60.0%和20.0%,2组之间差异有统计学意义(P<0.05)。胃癌组织中Skp2表达阳性率明显高于对照组(P<0.01);而RASSF1A表达阳性率明显低于对照组(P<0.01);H.pylori-L型阳性组的Skp2表达阳性率高于H.pylori-L型阴性组(P<0.05);H.pylori-L型阳性与Skp2的表达阳性呈正相关关系;RASSF1A的表达与H.pylori-L型阳性呈负相关关系。结论 H.pylori-L型感染在胃癌的发生发展过程中起重要的作用,其促进胃癌发生、发展的机制可能涉及上调Skp2的表达和下调RASSF1A的表达。     

BAIAP2L2在肝癌中的表达及临床意义

脑特异性血管生成抑制剂1相关蛋白2样2(brain-specific angiogenesis inhibitor 1-associated protein 2-like 2, BAIAP2L2)是一种上皮特异性BAR结构域蛋白,近年来被发现在多种肿瘤中高表达,然而在肝癌中的角色尚不清楚。本文通过生物信息学方法,探讨了BAIAP2L2在肝癌中的表达、生物学意义及潜在的临床应用价值,旨在为BAIAP2L2作为肝癌的新型标志物提供理论依据。文中首先从TCGA和ICGC数据库下载肝癌m RNA-seq、mi RNA-seq、DNA甲基化数据及临床病理资料,从HPA数据库获取蛋白质表达数据,以比较癌组织和癌旁组织中BAIAP2L2的表达差异,分析BAIAP2L2表达与肝癌临床病理特征的关系。随后,采用受试者工作特征(receiver operating characteristic, ROC)曲线评估BAIAP2L2表达水平在肝癌诊断中的性能,通过绘制生存曲线比较不同BAIAP2L2表达组患者的生存差异,并利用Cox回归计算风险比(hazard ratio, HR)。最后,利用mi RWalk...     

NFE2L1在食管鳞癌中的表达及临床意义

目的:通过检测食管鳞癌标本中核因子E2相关因子1(NFE2L1)的表达情况,探究NFE2L1在食管鳞癌发生发展中的作用,为食管鳞癌的诊治以及预后评估提供新的思路。方法:收集我校附院2016-2017年经病理组织学检查诊断为食管鳞癌的手术标本40例及其对应的癌旁组织并从NCBI数据库下载GEO测序数据,应用定量PCR、免疫组化和生物信息分析等方法检测分析NFE2L1基因的表达情况。结果:在食管鳞癌组织中,NFE2L1表达阳性31例(77.5%),癌旁组织阳性表达17例(42.5%),两组比较差异显著有统计学意义(P0.001);进一步发现NFE2L1的阳性表达与肿瘤分化程度和淋巴转移相关(P0.05)。但在不同年龄、性别、浸润深度及不同部位之间差异无统计学意义。GEO数据分析结果显示NFE2L1在食管鳞癌组织显著高表达(P0.01),只是未达到显著差异表达的阈值标准(即变化倍数小于2倍)。结论:NFE2L1在食管鳞癌中高表达,表达的高低与食管鳞癌的发生进展密切相关。     

运用组织芯片技术检测ndrg2、c-myc在人甲状腺肿瘤中的表达及分析

目的:检测ndrg2、c-myc在人甲状腺肿瘤中的表达,探讨ndrg2、c-myc在人甲状腺肿瘤中的表达模式.方法:运用甲状腺组织芯片,通过免疫组化染色,检测ndrg2、c-myc在人甲状腺肿瘤中的表达,并进行量化评分,分析ndrg2表达与c-myc表达两者之间可能存在的负调控关系.结果:ndrg2在甲状腺癌组织中呈阴性表达,在甲状腺腺瘤组织中呈弱表达或不表达趋势,在正常甲状腺组织中高表达.而c-myc在甲状腺癌组织中呈高表达,在甲状腺腺瘤组织中不表达或仅呈部分弱表达,在正常甲状腺组织中呈阴性表达.统计结果分析显示,ndrg2与c-myc在人甲状腺组织中的表达呈负相关.结论:ndrg2、c-myc在人甲状腺肿瘤中的表达呈负相关,由此推测NDRG2可能受到C-MYC的负调控.     

HMGA1通过Wnt/β-Catenin通路被诱导并维持胃癌细胞增殖

目的探讨HMGA1通过Wnt/β-Catenin通路在胃肿瘤形成中的作用。方法应用小干扰RNA介导的基因沉默、细胞增殖分析、PCR等技术完成实验。结果 HMGA1的特异敲除明显减少细胞生长。β-Catenin或下游c-myc的损失减少HMGA1表达,而Wnt3a处理增加HMGA1和c-myc的转录。结论这些数据表明,HMGA1参与胃癌细胞形成和增殖通过Wnt/β-Catenin通路。     

表皮生长因子及受体在甲状腺肿瘤中的表达

目的:研究表皮生长因子(Epidermal Growth Factor,EGF)及受体(Epidermal Growth Factor Receptor,EGFR)及在甲状腺肿瘤中的表达。方法:应用免疫组织化学法检测91例甲状腺病变组织中EGFR和EGF的表达情况。结果:结节性甲状腺肿、甲状腺腺瘤、分化型甲状腺癌标本中EGFR表达的阳性率分别为15%、25%、68.62%,EGF表达的阳性率分别为10%、15%、68.62%,其中EGFR、EGF在分化型甲状腺癌与其余两组间差异均有统计学意义(P<0.05)。EGFR和EGF在甲状腺乳头状癌中的表达与性别、年龄、肿瘤大小、淋巴结转移、临床分期等临床因素无明显相关。结论:EGF和EGFR的表达可作为鉴别甲状腺肿瘤良恶性的一个指标。     

HMGA1-pseudogene expression is induced in human pituitary tumors

Numerous studies have established that High Mobility Group A (HMGA) proteins play a pivotal role on the onset of human pituitary tumors. They are overexpressed in pituitary tumors, and, consistently, transgenic mice overexpressing either the Hmga1 or the Hmga2 gene develop pituitary tumors. In contrast with HMGA2, HMGA1 overexpression is not related to any rearrangement or amplification of the HMGA1 locus in these tumors. We have recently identified 2 HMGA1 pseudogenes, HMGA1P6 and HMGA1P7, acting as competitive endogenous RNA decoys for HMGA1 and other cancer related genes. Here, we show that HMGA1 pseudogene expression significantly correlates with HMGA1 mRNA levels in growth hormone and nonfunctioning pituitary adenomas likely inhibiting the repression of HMGA1 through microRNAs action. According to our functional studies, these HMGA1 pseudogenes enhance the proliferation and migration of the mouse pituitary tumor cell line, at least in part, through their upregulation. Our results point out that the overexpression of HMGA1P6 and HMGA1P7 could contribute to increase HMGA1 levels in human pituitary tumors, and then to pituitary tumorigenesis.     

Pseudogene HSPB1P1 contributes to renal cell carcinoma proliferation and metastasis by targeting miR-296-5p to regulate HMGA1 expression

With the aid of next-generation sequencing technology, pseudogenes have been widely recognized as functional regulators in the development and progression of certain diseases, especially cancer. Our present study aimed to investigate the functions and molecular mechanisms of HSPB1-associated protein 1 pseudogene 1 (HSPB1P1) in renal cell carcinoma (RCC). HSPB1P1 expression at the mRNA levels was determined by quantitative real-time polymerase chain reaction, and its clinical significance was assessed. Cell viability was detected by Cell Counting Kit-8 assay. Cell migration and invasion were detected by transwell assays. The location of HSPB1P1 in RCC cells was detected by subcellular distribution analysis. The direct relationship between HSPB1P1 and miR-296-5p/HMGA1 axis was verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. Our results identify the elevated expression of HSPB1P1 in RCC tissues and cell lines, which predicted advanced progression and poor prognosis in patients with RCC. Knockdown of HSPB1P1 suppressed cell proliferation, migration, and invasion, and reversed epithelial–mesenchymal transition process in RCC. HSPB1P1 was mostly enriched in the cytoplasm and functioned as a miRNA sponge for miR-296-5p and then regulated high-mobility group A1 expression. In conclusion, our study indicated that HSPB1P1 contributed to RCC progression by targeting the miR-296-5p/HMGA1 axis, and should be considered as a promising biomarker and therapeutic target for clinical applications.     

The nuclear architectural protein HMGA1a triggers receptor‐mediated endocytosis

High mobility group proteins A (HMGA), nuclear architectural factors, locate in the cell nuclei and mostly execute gene‐regulation function. However, our results reveal that a HMGA member (HMGA1a) has a unique plasma membrane receptor; this receptor specifically binds to HMGA‐decorated species, effectively mediates endocytosis, and internalizes extracellular HMGA‐functionalized cargoes. Indeed, dyes or nanoparticles labeled with HMGA1a protein readily enter Hela cells. Using a stratagem chemical cross‐linker, we covalently bonded the HMGA receptor to the HMGA1a‐GFP fusion protein, thus capturing the plasma membrane receptor. Subsequent Western blots and SDS–PAGE gel revealed that the HMGA receptor is a 26‐kDa protein. Confocal live‐cell microscopic imaging was used to monitor the whole endocytic process, in which the internalized HMGA1a‐decorated species are transported by motor proteins on microtubules and eventually arrive at the late endosomes/lysosomes. Cell viability assays also suggested that extracellular HMGA1a protein directly influences the survival ability of Hela cells in a dose‐dependent manner, implying versatility of HMGA1a protein and its potent role to suppress cancer cell survivability and to regulate growth. J. Cell. Biochem. 108: 791–801, 2009. © 2009 Wiley‐Liss, Inc.     

NP c 1L l 在高脂血症和动脉粥样硬化中的表达分析

目的：研究NPC1L1（Niemann-Pick C1 Like 1）mRNA在单纯高脂血症大鼠和动脉粥样硬化大鼠小肠组织中的表达与差异,探讨其与脂质代谢和动脉粥样硬化之间的关系。方法：通过半定量RT-PCR方法分别检测正常普食组、单纯高脂饲养组和动脉粥样大鼠组小肠组织中NPC1L1 mRNA的表达差异。结果：三个组别大鼠小肠组织中均存在NPC1L2 mRNA,单纯高脂饮食和动脉粥样大鼠小肠组织中NPC1L1 mRNA表达明显高于正常对照大鼠（P〈0．01）;单纯高脂饮食和动脉粥样大鼠小肠组织中NPC1L1 mRNA表达之间无明显差异（P〉0．05）。结论：血脂代谢紊乱与小肠组织中NPC1L1的高表达有关,NPC1L1可能参与了血脂紊乱的病理生理过程;NPC1L1与促成动脉粥样硬化的发生无明显相关性。     

The energetics of specific binding of AT-hooks from HMGA1 to target DNA

The interaction of the second and third AT-hooks of HMGA1 (formerly HMGI/Y), which bind selectively in the minor groove of an AT-rich DNA sequence, was studied at different temperatures and ionic strengths by spectropolarimetry, spectrofluorimetry, isothermal titration calorimetry and differential scanning calorimetry. The data show that binding of the ten amino acid core element of the two AT-hooks, which penetrates deep into the minor groove, is entropically driven: both the entropy and enthalpy of association of the peptides to the target DNA are positive up to 50 degrees C. The seven amino acid extension of the core in the second AT-hook, which extends out from the minor groove and loops over the phosphodiester backbone, adds a substantial negative enthalpic component into the binding of the 17 residue DBD2 peptide to DNA that corresponds in magnitude to the enthalpy of formation of two hydrogen bonds. The ionic strength dependence of the association constant allowed an estimation of the electrostatic component of binding and, by subtraction, the contribution of the non-electrostatic component, which results from dehydration of the contacting surfaces and makes up almost 70% of the total energy of complex formation. The exceptionally large positive entropy and enthalpy of association of the core AT-hook peptides with target DNA suggest that the water, which is removed from the minor groove of DNA upon binding, is in a highly ordered state. Acetylation of the lysine residue in the second AT-hook, which corresponds to Lys65 of HMGA1, has little effect on the DNA binding; so it appears that repression of the hIFNbeta gene, which follows this modification, is not a direct result of the abrogation of DNA binding.