Background::Pancreatic cancer (PC) is a highly deadly malignancy with few effective therapies. We aimed to unmask the role that long non-coding RNA small nucleolar RNA host gene 6 ( SNHG6) plays in PC cells by targeting far upstream element binding protein 1 ( FUBP1) via microRNA-26a-5p ( miR-26a-5p). Methods::SNHG6 expression was predicted by bioinformatics, followed by verification via reverse transcription quantitative polymerase chain reaction. Then, the interactions among SNHG6, miR-26a-5p, and FUBP1 were detected through online software analysis, dual luciferase reporter assay and RNA pull-down. After that, cells were treated with different small interfering RNAs and/or mimic to determine the interactions among SNHG6, miR-26a-5p, and FUBP1 and their roles in PC cells. Finally, the role of SNHG6 in tumor growth in vivo was evaluated by measuring the growth and weight of transplanted tumors in nude mice. A t-test, one-way and two-way analysis of variance were used for data analysis. Results::Compared with that in normal tissues, SNHG6 was highly expressed in PC tissues (1.00 ± 0.05 vs. 1.56 ± 0.06, t= 16.03, P < 0.001). Compared with that in human pancreatic duct epithelial cells (HPDE6-C7), SNHG6 showed the highest expression in PANC-1 cells (1.00 ± 0.06 vs. 3.87 ± 0.13, t= 34.72, P < 0.001) and the lowest expression in human pancreatic cancer cells (MIAPaCa-2) (1.00 ± 0.06 vs. 1.41 ± 0.07, t= 7.70, P= 0.0015). Compared with the levels in the si-negative control group, SNHG6 (0.97 ± 0.05 vs. 0.21 ± 0.06, t = 16.85, P < 0.001), N-cadherin (0.74 ± 0.05 vs. 0.41 ± 0.04, t= 8.93, P < 0.001), Vimentin (0.55 ± 0.04 vs. 0.25 ± 0.03, t= 10.39, P < 0.001), and β-catenin (0.62 ± 0.05 vs . 0.32 ± 0.03, t= 8.91, P < 0.001) were decreased, while E-cadherin (0.65 ± 0.06 vs. 1.36 ± 0.07, t= 13.34, P < 0.001) was increased after SNHG6 knockdown or miR-26a-5p overexpression, accompanied by inhibited cell proliferation, migration, and invasion. SNHG6 overexpression exerted the opposite effects. SNHG6 upregulated FUBP1 expression by sponging miR-26a-5p. Silencing SNHG6 blocked the growth of PC in vivo. Conclusion::Silencing SNHG6 might ameliorate PC through inhibition of FUBP1 by sponging miR-26a-5p, thus providing further supporting evidence for its use in PC treatment.

目的 探究远端上游元件结合蛋白3(far upstream element binding protein 3,FUBP3)介导糖酵解对成骨细胞分化能力和功能的影响.方法 实时荧光定量PCR(reverse transcription real-time fluorescence quantitative PCR,RT-qPCR)检测骨质疏松性骨折患者外周血和接受入院 24 h、接受治疗 1、2、3、4 周时FUBP3 mRNA的表达水平,RT-qPCR和蛋白免疫印迹法(Western blot)检测骨质疏松性骨折患者骨组织FUBP3 mRNA和蛋白表达水平.MC3T3-E1 细胞分为sh-NC组、sh-FUBP3 组、pcDNA组、pcDNA-FUBP3 组,MTT和EdU检测细胞增殖活性,Western blot检测Runt相关转录因子 2(runtrelated transcription factor 2,Runx2)、骨桥蛋白(osteopontin,OPN)、葡萄糖转运蛋白1(glucose transporters type 1,Glut1)、Glut3 蛋白,试剂盒检测碱性磷酸酶(alkaline phosphatse,ALP)活性、葡萄糖消耗量及乳酸生成量.结果 骨质疏松性骨折患者外周血中FUBP3 mRNA表达水平高于非骨质疏松性骨折患者,骨质疏松性骨折患者骨组织中FUBP3 mRNA和蛋白表达水平高于非骨质疏松性骨折患者(P<0.05);骨质疏松性骨折患者在入院 24 h、接受治疗 1、2、3、4 周时外周血,发现随着治疗时间增加FUBP3 mRNA逐渐降低,呈时间依赖性,在治疗4 周时表达水平最低(P<0.05).与sh-NC组相比,sh-FUBP3 组FUBP3 mRNA和蛋白表达明显降低,细胞活性和EdU阳性细胞比率、葡萄糖消耗量、乳酸生成量、ALP 活性明显增加,Glut1、Glut3、Runx2、OPN 蛋白表达上调(P<0.05);与pcDNA组相比,pcDNA-FUBP3 组FUBP3 mRNA和蛋白表达明显增加,细胞活性和EdU阳性细胞比率、葡萄糖消耗量、乳酸生成量、ALP活性明显降低,Glut1、Glut3、Runx2、OPN蛋白表达下调(P<0.05).结论 骨质疏松性骨折患者外周血和骨组织中FUBP3 表达上调,敲低FUBP3 抑制成骨细胞增殖、成骨分化和糖酵解,过表达FUBP3 则相反.

Glioma is a complex disease with limited treatment options.Recent advances have identified isocitrate dehydrogenase (IDH)mutations in up to 80％ lower grade gliomas (LGG) and in 76％ secondary glioblastomas (GBM).IDH mutations are also seen in 10％-20％ of acute myeloid leukemia (AML).In AML,it was determined that mutations of IDH and other genes involving epigenetic regulations are early events,emerging in the pre-leukemic stem cells (pre-LSCs) stage,whereas mutations in genes propagating oncogenic signal are late events in leukemia.IDH mutations are also early events in glioma,occurring before TP53 mutation,1p/19q deletion,etc.Despite these advances in glioma research,studies into other molecular alterations have lagged considerably.In this study,we analyzed currently available databases.We identified EZH2,KMT2C,and CHD4 as important genes in glioma in addition to the known gene IDH1/2.We also showed that genomic alterations of PIK3CA,CDKN24,CDK4,FIP1L1,or FUBP1 collaborate with IDH mutations to negatively affect patients' survival in LGG.In LGG patients with TP53 mutations or IDH1/2 mutations,additional genomic alterations of EZH2,KMC2C,and CHD4 individually or in combination were associated with a markedly decreased disease-free survival than patients without such alterations.Alterations of EZH2,KMT2C,and CHD4 at genetic level or protein level could perturb epigenetic program,leading to malignant transformation in glioma.By reviewing current literature on both AML and glioma and performing bioinformatics analysis on available datasets,we developed a hypothetical model on the tumorigenesis from premalignant stem cells to glioma.

[目的]研究人远端上游原件结合蛋白(FUBP1)在舌鳞状细胞癌(TSCC)组织及其癌旁正常组织中的表达情况,探讨FUBP1在TSCC发生发展中的作用及其临床意义.[方法]采用免疫组化法检测FUBP1在收集的90例TSCC组织及其癌旁正常组织中的表达;qRT-PCR及Western blot技术分别检测20对和8对TSCC组织及其癌旁正常组织中FUBP1的表达,并分析其与患者临床病理参数、预后的关系.[结果]免疫组化、qRT-PCR和Western blot结果均显示,TSCC组织中FUBP1的表达水平高于相应癌旁正常组织(P＜0.05);TSCC组织标本中的FUBP1在晚期、颈淋巴结转移患者中呈高表达(P＜0.001),FUBP1高表达组比FUBP1低表达组的5年生存率低(P=0.035);FUBP1表达水平是影响患者总生存率的独立预测指标.[结论]FUBP1在TSCC组织中的表达上调并且与TSCC患者不良预后关系密切,提示FUBP1高表达可能促进TSCC肿瘤形成和发展,并作为判断患者预后的指标.

目的 研究远端上游元件结合蛋白1(far upstream element-binding protein 1,FUBP1)在幽门螺杆菌(Helicobacter pylori,H.pylori)相关性胃炎中的表达,并初步探讨H.pylori引起FUBP1表达变化的可能机制.方法 分别采用实时荧光定量逆转录聚合酶链式反应(RT-qPCR)、蛋白质印迹法(Western blotting)和免疫组织化学技术检测H.pylori阴性和H.pylori阳性胃炎组织中FUBP1 mRNA和蛋白的表达.国际标准测序株H.pylori 26695及其提取的LPS分别刺激AGS、SGC7901细胞,RT-qPCR和Western blotting检测FUBP1 mRNA和蛋白的表达.分别使用H.pylori和H.pylori-LPS灌胃,构建小鼠感染模型,于灌胃结束后4周、8周取小鼠胃黏膜组织,RT-qPCR和Western blotting检测FUBP1 mRNA和蛋白表达.结果 H.pylori阳性组织中FUBP1 mRNA及蛋白表达水平均低于H.pylori阴性组织,FUBP1蛋白水平的表达差异有统计学意义(P<0.05).H.pylori刺激AGS和SGC7901细胞后FUBP1 mRNA和蛋白水平降低,且呈时间依赖性,H.pylori-LPS刺激细胞后,FUBP1 mRNA和蛋白水平表达降低,但无时间及浓度依赖性.动物模型中,与对照组相比,H.pylori组和H.pylori-LPS组的FUBP1 mRNA和蛋白表达水平均明显降低,差异有统计学意义(P<0.05).结论 FUBP1在H.pylori相关性胃炎中表达下降,H.pylori和H.pylori毒力因子LPS均能诱导FUBP1下调,FUBP1可能成为评价慢性胃炎患者H.pylori感染的一个潜在指标.

远端上游元件结合蛋白1(FUBP1)是一种单链核酸结合蛋白,由有螺旋结构的N端、富含酪氨酸的C端和一个DNA结合区域构成,与远端上游元件(FUSE)结合后可调控原癌基因c-Myc的转录.FUBP1通过调控基因转录、翻译、RNA复制和剪接,影响细胞增殖、迁移、凋亡等过程,在多种肿瘤的发生、发展中扮演重要角色.本文就FUBP1的结构、功能、家族成员及其与消化道肿瘤相关机制的研究作一综述.

目的 对Xp11.2易位/TFE3基因不同融合类型设计多重PCR引物,证实多重PCR联合Sanger法测序的可行性,旨在探讨引物设计的重要性.方法 选取已通过高通量RNA-Seq测序证实TFE3融合类型的标本23例,其融合类型包括PSF-TFE3 3种,PRCC-TFE3 3种,ASPL-TFE3 2种, FUBP1-TFE3 1种,NONO-TFE3 2种,RBM10-TFE3 1种,MED15-TFE3 1种以及MATR3-TFE3 1种共计14种,对其用设计好的多重PCR引物进行PCR扩增并联合Sanger法测序进行验证,该次实验分4个多重PCR反应,包括14条不同基因的正向引物以及4条TFE3反向引物.结果 23例标本均与高通量RNA-Seq测序结果完全一致,可见所设计的引物通过多重PCR联合Sanger法测序是可行的.结论该方法的优势在于利用4个多重PCR反应便可确定是否存在上述14种TFE3融合类型阳性,并通过反向测序便可确定具体融合类型.因此从试剂成本方面考虑,尤其在大批量标本的TFE3融合类型筛查过程中,该方法较高通量RNA-Seq测序有很大的优势.

目的 应用生物信息学方法 分析hsa-miR-206序列,并进行靶基因功能富集分析、信号通路富集分析,靶基因编码蛋白相互作用分析,为后续研究其功能提供理论基础.方法 通过miRbase、TargetScan、DIANA-microT、MiRDB据库、VENN在线工具、DAVID在线数据库、STRING数据库及Cytoscape软件等在线工具分析hsa-miR-206序列及保守性,预测其靶基因,对预测的靶基因进行功能富集分析(GO分析)、KEGG通路富集分析,并筛选出关键基因.结果 hsa-miR-206序列在各物种间高度保守,GO分析发现hsa-miR-206靶基因功能主要富集在生物合成调节、有机物代谢过程调节、转录调节等生物学过程,KEGG分析表明主要参与Wnt信号通路、T细胞受体信号通路、癌症通路等.蛋白互作显示编码蛋白间存在复杂的相互作用,与hsa-miR-206关系最密切的蛋白有27个,如天冬氨酸-谷氨酸-丙氨酸-天冬氨酸盒解螺旋酶5(DDX5)、人远端上游原件结合蛋白(FUBP1)、天冬氨酸-谷氨酸-丙氨酸-组氨酸盒解螺旋酶15(DHX15)等.结论 hsa-miR-206可能参与肿瘤发生发展中重要的生物学过程及调控机制,为进一步实验验证提供了线索.

目的:研究敲低远端上游元件结合蛋白1(FUBP1)的表达对胰腺癌细胞增殖和侵袭迁移能力的影响.方法:细胞转染干扰序列siFUBP1降低胰腺癌细胞PaTu8988中FUBP1的表达,FUBP1-nc作为阴性对照.CCK-8实验检测细胞增殖能力,细胞划痕实验和Transwell实验检测细胞侵袭迁移能力.结果:敲低胰腺癌细胞PaTu8988中FUBP1的表达能明显抑制细胞增殖,细胞侵袭迁移数目减少,降低细胞侵袭迁移能力.结论:敲低FUBP1的表达能显著抑制胰腺癌细胞增殖及侵袭迁移能力.

目的 探讨人远端上游原件结合蛋白(FUBP1)在宫颈癌中的表达水平及其与预后的相关性.方法 回顾性选取2016年1月至2021年5月在同济大学附属第一妇婴保健院妇科肿瘤病房接受治疗的宫颈癌患者,获取宫颈癌组织和癌旁组织86例.采用免疫组织化学方法检测FUBP1的表达,按照中位数法进行分组,FUBP1蛋白>0.83为FUBP1高表达组,FUBP1蛋白≤0.83为FUBP1低表达组.分析FUBP1表达与临床病理的相关性,采用受试者工作特性(ROC)曲线分析FUBP1在宫颈癌中的诊断价值,采用Kaplan-Meier法绘制生存曲线,对比FUBP1高表达组和低表达组患者的总生存率,采用Cox比例风险模型进行单因素和多因素分析.结果 FUBP1在癌旁组织和宫颈癌组织中表达分别为0.27±0.03和1.48±0.11,FUBP1在宫颈癌组织中的表达明显高于癌旁组织,差异有统计学意义(P<0.001).FUBP1诊断宫颈癌的ROC曲线下面积(AUC)为0.921(95％CI:0.882～0.962,P<0.001),最佳截断值为1.305,敏感度为79.40％,特异度为68.78％.FUBP1高表达组的总生存率显著低于FUBP1低表达组,差异有统计学意义(P<0.05);FIGO分期、淋巴结转移、宫颈浸润深度、FUBP1的表达与宫颈癌患者预后有关(P<0.05);FIGO分期Ⅲ-Ⅳ期、有淋巴结转移、宫颈浸润深度≥2/3、FUBP1高表达是宫颈癌预后独立危险因素(HR=6.396,95％CI:4.116～15.279,P<0.001;HR=5.173,95％CI:1.936～12.187,P<0.001;HR=4.518,95％CI:1.163～11.729,P<0.001;HR=5.149,95％CI:2.073～13.272,P<0.001).结论 FUBP1在宫颈癌中呈现高表达,检测宫颈癌组织中FUBP1的表达水平对宫颈癌的诊断及预后判断有重要的临床价值.