BACKGROUND & AIMS: :Telomeres are nucleoprotein structures at the ends of linear chromosomes and present an essential feature for genome integrity. Vertebrate telomeres usually consist of hexameric TTAGGG repeats, however, in cells that use the alternative lengthening of telomeres (ALT) mechanism, variant repeat sequences are interspersed throughout telomeres. Previously, it was shown that NR2C/F transcription factors bind to TCAGGG variant repeats and contribute to telomere maintenance in ALT cells. While specific binders to other variant repeat sequences have been lacking to date, we here identify ZBTB10 as the first TTGGGG-binding protein and demonstrate direct binding via the two zinc fingers with affinity in the nanomolar range. Concomitantly, ZBTB10 co-localizes with a subset of telomeres in ALT-positive U2OS cells and interacts with TRF2/RAP1 via the N-terminal region of TRF2. Our data establishes ZBTB10 as a novel variant repeat binding protein at ALT telomeres.

背景与目标： ：端粒是线性染色体末端的核蛋白结构，具有基因组完整性的重要特征。脊椎动物端粒通常由六聚体TTAGGG重复序列组成，但是，在使用端粒交替延长（ALT）机制的细胞中，变异重复序列散布在整个端粒中。以前，已显示NR2C / F转录因子与TCAGGG变体重复结合并有助于ALT细胞中端粒的维持。尽管迄今为止还缺乏针对其他变异重复序列的特异性结合物，但我们在这里将ZBTB10确定为第一个TTGGGG结合蛋白，并证明通过两个锌指直接结合，其亲和力在纳摩尔范围内。同时，ZBTB10与ALT阳性U2OS细胞中的一部分端粒共定位，并通过TRF2的N端区域与TRF2 / RAP1相互作用。我们的数据将ZBTB10建立为ALT端粒的新型变异重复结合蛋白。

BACKGROUND & AIMS: :To investigate the genetics of late-onset myasthenia gravis (LOMG), we conducted a genome-wide association study imputation of>6 million single nucleotide polymorphisms (SNPs) in 532 LOMG cases (anti-acetylcholine receptor [AChR] antibody positive; onset age≥50 years) and 2,128 controls matched for sex and population substructure. The data confirm reported TNFRSF11A associations (rs4574025, P = 3.9 × 10-7, odds ratio [OR] 1.42) and identify a novel candidate gene, ZBTB10, achieving genome-wide significance (rs6998967, P = 8.9 × 10-10, OR 0.53). Several other SNPs showed suggestive significance including rs2476601 (P = 6.5 × 10-6, OR 1.62) encoding the PTPN22 R620W variant noted in early-onset myasthenia gravis (EOMG) and other autoimmune diseases. In contrast, EOMG-associated SNPs in TNIP1 showed no association in LOMG, nor did other loci suggested for EOMG. Many SNPs within the major histocompatibility complex (MHC) region showed strong associations in LOMG, but with smaller effect sizes than in EOMG (highest OR ~2 versus ~6 in EOMG). Moreover, the strongest associations were in opposite directions from EOMG, including an OR of 0.54 for DQA1*05:01 in LOMG (P = 5.9 × 10-12) versus 2.82 in EOMG (P = 3.86 × 10-45). Association and conditioning studies for the MHC region showed three distinct and largely independent association peaks for LOMG corresponding to (a) MHC class II (highest attenuation when conditioning on DQA1), (b) HLA-A and (c) MHC class III SNPs. Conditioning studies of human leukocyte antigen (HLA) amino acid residues also suggest potential functional correlates. Together, these findings emphasize the value of subgrouping myasthenia gravis patients for clinical and basic investigations and imply distinct predisposing mechanisms in LOMG.

背景与目标： ：为了研究迟发性重症肌无力（LOMG）的遗传学，我们进行了全基因组关联研究，估算了532例LOMG病例中> 600万个单核苷酸多态性（SNP）（抗乙酰胆碱受体[AChR]抗体呈阳性；年龄≥50岁）和2,128名对照者的性别和人口子结构相匹配。数据证实了已报道的TNFRSF11A关联（rs4574025，P = 3.9×10-7，比值比[OR] 1.42），并鉴定了一个新的候选基因ZBTB10，实现了全基因组意义（rs6998967，P = 8.9×10-10，或0.53）。其他几个SNP也显示出提示意义，包括rs2476601（P = 6.5×10-6，或1.62），编码早发性重症肌无力（EOMG）和其他自身免疫性疾病中提到的PTPN22 R620W变体。相比之下，TNIP1中与EOMG相关的SNP在LOMG中未显示任何关联，也未建议对EOMG使用其他基因座。主要组织相容性复合物（MHC）区域内的许多SNP在LOMG中显示出很强的关联性，但其效应大小比EOMG小（在EOMG中，最高OR〜2对〜6）。此外，最强的关联与EOMG方向相反，包括LOMG中DQA1 * 05：01的OR为0.54（P = 5.9×10-12），而EOMG中为2.82（P = 3.86×10-45）。 MHC区域的关联和条件研究显示，LOMG的三个不同且基本独立的关联峰分别对应于（a）II类MHC（对DQA1进行条件调节时衰减最大），（b）HLA-A和（c）III类MHC SNP。人类白细胞抗原（HLA）氨基酸残基的条件研究也表明潜在的功能相关性。总之，这些发现强调了重症肌无力患者亚组在临床和基础研究中的价值，并暗示了LOMG的独特诱因机制。

BACKGROUND & AIMS:

背景与目标：

BACKGROUND & AIMS: :Several studies have demonstrated that polyphenolics from pomegranate (Punica granatum L.) are potent inhibitors of cancer cell proliferation and induce apoptosis, cell cycle arrest, and also decrease inflammation in vitro and vivo. There is growing evidence that botanicals exert their cytotoxic and anti-inflammatory activities, at least in part, by decreasing specificity protein (Sp) transcription factors. These are overexpressed in breast tumors and regulate genes important for cancer cell survival and inflammation such as the p65 unit of NF-κB. Moreover, previous studies have shown that Pg extracts decrease inflammation in lung cancer cell lines by inhibiting phosphatidylinositol-3,4,5-trisphosphate (PI3K)-dependent phosphorylation of AKT in vitro and inhibiting the activation of NF-kB in vivo. The objective of this study was to investigate the roles of miR-27a-ZBTB10-Sp and miR-155-SHIP-1-PI3K on the anti-inflammatory and cytotoxic activity of pomegranate extract. Pg extract (2.5-50 μg/ml) inhibited growth of BT-474 and MDA-MB-231 cells but not the non-cancer MCF-10F and MCF-12F cells. Pg extract significantly decreased Sp1, Sp3, and Sp4 as well as miR-27a in BT474 and MDA-MB-231 cells and increased expression of the transcriptional repressor ZBTB10. A significant decrease in Sp proteins and Sp-regulated genes was also observed. Pg extract also induced SHIP-1 expression and this was accompanied by downregulation of miRNA-155 and inhibition of PI3K-dependent phosphorylation of AKT. Similar results were observed in tumors from nude mice bearing BT474 cells as xenografts and treated with Pg extract. The effects of antagomirs and knockdown of SHIP-1 by RNA interference confirmed that the anti-inflammatory and cytotoxic effects of Pg extract were partly due to the disruption of both miR-27a-ZBTB10 and miR-155-SHIP-1. In summary, the anticancer activities of Pg extract in breast cancer cells were due in part to targeting microRNAs155 and 27a. Both pathways play an important role in the proliferative/inflammatory phenotype exhibited by these cell lines.

背景与目标： ：几项研究表明，石榴中的多酚类物质（Punica granatum L.）是癌细胞增殖的有效抑制剂，可诱导癌细胞凋亡，细胞周期停滞并在体内和体外减少炎症。越来越多的证据表明，植物药至少部分地通过降低特异性蛋白（Sp）转录因子来发挥其细胞毒性和抗炎活性。这些在乳腺肿瘤中过表达，并调节对于癌细胞存活和炎症重要的基因，例如NF-κB的p65单元。此外，以前的研究表明，Pg提取物通过在体外抑制AKT的磷脂酰肌醇-3,4,5-三磷酸（PI3K）依赖性磷酸化并在体内抑制NF-kB的活化来减轻肺癌细胞的炎症。这项研究的目的是调查miR-27a-ZBTB10-Sp和miR-155-SHIP-1-PI3K对石榴提取物的抗炎和细胞毒性活性的作用。 Pg提取物（2.5-50μg/ ml）抑制BT-474和MDA-MB-231细胞的生长，但不抑制非癌MCF-10F和MCF-12F细胞的生长。 Pg提取物显着降低BT474和MDA-MB-231细胞中的Sp1，Sp3和Sp4以及miR-27a，并增加转录阻遏物ZBTB10的表达。还观察到Sp蛋白和Sp调节基因的显着减少。 Pg提取物也诱导SHIP-1表达，并伴随miRNA-155的下调和PI3K依赖性AKT磷酸化的抑制。在携带BT474细胞作为异种移植物并用Pg提取物治疗的裸鼠的肿瘤中观察到了相似的结果。 RNA干扰的拟南芥作用和SHIP-1的敲除证实了Pg提取物的抗炎和细胞毒性作用部分是由于miR-27a-ZBTB10和miR-155-SHIP-1的破坏。总之，Pg提取物在乳腺癌细胞中的抗癌活性部分归因于靶向microRNAs155和27a。两种途径在这些细胞系表现出的增殖/炎性表型中都起着重要作用。

BACKGROUND & AIMS: :Telomeres are nucleoprotein structures at the ends of linear chromosomes and present an essential feature for genome integrity. Vertebrate telomeres usually consist of hexameric TTAGGG repeats, however, in cells that use the alternative lengthening of telomeres (ALT) mechanism, variant repeat sequences are interspersed throughout telomeres. Previously, it was shown that NR2C/F transcription factors bind to TCAGGG variant repeats and contribute to telomere maintenance in ALT cells. While specific binders to other variant repeat sequences have been lacking to date, we here identify ZBTB10 as the first TTGGGG-binding protein and demonstrate direct binding via the two zinc fingers with affinity in the nanomolar range. Concomitantly, ZBTB10 co-localizes with a subset of telomeres in ALT-positive U2OS cells and interacts with TRF2/RAP1 via the N-terminal region of TRF2. Our data establishes ZBTB10 as a novel variant repeat binding protein at ALT telomeres.

背景与目标： ：端粒是线性染色体末端的核蛋白结构，具有基因组完整性的重要特征。脊椎动物端粒通常由六聚体TTAGGG重复序列组成，但是，在使用端粒交替延长（ALT）机制的细胞中，变异重复序列散布在整个端粒中。以前，已显示NR2C / F转录因子与TCAGGG变体重复结合并有助于ALT细胞中端粒的维持。尽管迄今为止还缺乏针对其他变异重复序列的特异性结合物，但我们在这里将ZBTB10确定为第一个TTGGGG结合蛋白，并证明通过两个锌指直接结合，其亲和力在纳摩尔范围内。同时，ZBTB10与ALT阳性U2OS细胞中的一部分端粒共定位，并通过TRF2的N端区域与TRF2 / RAP1相互作用。我们的数据将ZBTB10建立为ALT端粒的新型变异重复结合蛋白。

BACKGROUND & AIMS: :To investigate the genetics of late-onset myasthenia gravis (LOMG), we conducted a genome-wide association study imputation of>6 million single nucleotide polymorphisms (SNPs) in 532 LOMG cases (anti-acetylcholine receptor [AChR] antibody positive; onset age≥50 years) and 2,128 controls matched for sex and population substructure. The data confirm reported TNFRSF11A associations (rs4574025, P = 3.9 × 10-7, odds ratio [OR] 1.42) and identify a novel candidate gene, ZBTB10, achieving genome-wide significance (rs6998967, P = 8.9 × 10-10, OR 0.53). Several other SNPs showed suggestive significance including rs2476601 (P = 6.5 × 10-6, OR 1.62) encoding the PTPN22 R620W variant noted in early-onset myasthenia gravis (EOMG) and other autoimmune diseases. In contrast, EOMG-associated SNPs in TNIP1 showed no association in LOMG, nor did other loci suggested for EOMG. Many SNPs within the major histocompatibility complex (MHC) region showed strong associations in LOMG, but with smaller effect sizes than in EOMG (highest OR ~2 versus ~6 in EOMG). Moreover, the strongest associations were in opposite directions from EOMG, including an OR of 0.54 for DQA1*05:01 in LOMG (P = 5.9 × 10-12) versus 2.82 in EOMG (P = 3.86 × 10-45). Association and conditioning studies for the MHC region showed three distinct and largely independent association peaks for LOMG corresponding to (a) MHC class II (highest attenuation when conditioning on DQA1), (b) HLA-A and (c) MHC class III SNPs. Conditioning studies of human leukocyte antigen (HLA) amino acid residues also suggest potential functional correlates. Together, these findings emphasize the value of subgrouping myasthenia gravis patients for clinical and basic investigations and imply distinct predisposing mechanisms in LOMG.

背景与目标： ：为了研究迟发性重症肌无力（LOMG）的遗传学，我们进行了全基因组关联研究，估算了532例LOMG病例中> 600万个单核苷酸多态性（SNP）（抗乙酰胆碱受体[AChR]抗体呈阳性；年龄≥50岁）和2,128名对照者的性别和人口子结构相匹配。数据证实了已报道的TNFRSF11A关联（rs4574025，P = 3.9×10-7，比值比[OR] 1.42），并鉴定了一个新的候选基因ZBTB10，实现了全基因组意义（rs6998967，P = 8.9×10-10，或0.53）。其他几个SNP也显示出提示意义，包括rs2476601（P = 6.5×10-6，或1.62），编码早发性重症肌无力（EOMG）和其他自身免疫性疾病中提到的PTPN22 R620W变体。相比之下，TNIP1中与EOMG相关的SNP在LOMG中未显示任何关联，也未建议对EOMG使用其他基因座。主要组织相容性复合物（MHC）区域内的许多SNP在LOMG中显示出很强的关联性，但其效应大小比EOMG小（在EOMG中，最高OR〜2对〜6）。此外，最强的关联与EOMG方向相反，包括LOMG中DQA1 * 05：01的OR为0.54（P = 5.9×10-12），而EOMG中为2.82（P = 3.86×10-45）。 MHC区域的关联和条件研究显示，LOMG的三个不同且基本独立的关联峰分别对应于（a）II类MHC（对DQA1进行条件调节时衰减最大），（b）HLA-A和（c）III类MHC SNP。人类白细胞抗原（HLA）氨基酸残基的条件研究也表明潜在的功能相关性。总之，这些发现强调了重症肌无力患者亚组在临床和基础研究中的价值，并暗示了LOMG的独特诱因机制。

BACKGROUND & AIMS: :Reactive oxygen species (ROS)-inducing anticancer agents such as phenethylisothiocyanate (PEITC) activate stress pathways for killing cancer cells. Here we demonstrate that PEITC-induced ROS decreased expression of microRNA 27a (miR-27a)/miR-20a:miR-17-5p and induced miR-regulated ZBTB10/ZBTB4 and ZBTB34 transcriptional repressors, which, in turn, downregulate specificity protein (Sp) transcription factors (TFs) Sp1, Sp3, and Sp4 in pancreatic cancer cells. Decreased expression of miR-27a/miR-20a:miR-17-5p by PEITC-induced ROS is a key step in triggering the miR-ZBTB Sp cascade leading to downregulation of Sp TFs, and this is due to ROS-dependent epigenetic effects associated with genome-wide shifts in repressor complexes, resulting in decreased expression of Myc and the Myc-regulated miRs. Knockdown of Sp1 alone by RNA interference also induced apoptosis and decreased pancreatic cancer cell growth and invasion, indicating that downregulation of Sp transcription factors is an important common mechanism of action for PEITC and other ROS-inducing anticancer agents.

背景与目标： ：诱导活性氧（ROS）的抗癌剂，例如苯乙基异硫氰酸酯（PEITC）激活了用于杀死癌细胞的应激途径。在这里，我们证明了PEITC诱导的ROS降低了microRNA 27a（miR-27a）/ miR-20a：miR-17-5p的表达并诱导了miR调节的ZBTB10 / ZBTB4和ZBTB34转录阻遏物，从而反过来下调了特异性蛋白（ Sp）胰腺癌细胞中的转录因子（TFs）Sp1，Sp3和Sp4。 PEITC诱导的ROS降低miR-27a / miR-20a：miR-17-5p的表达是触发miR-ZBTB Sp级联反应导致Sp TF下调的关键步骤，这是由于ROS依赖的表观遗传效应与阻遏物复合物的全基因组转移相关，导致Myc和Myc调节的miR的表达下降。通过RNA干扰单独击倒Sp1也会诱导凋亡，并降低胰腺癌细胞的生长和侵袭，这表明Sp转录因子的下调是PEITC和其他诱导ROS的抗癌药物的重要重要共同作用机制。

BACKGROUND & AIMS: :The anticancer agent 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) and its methyl ester (CDDO-Me) typically induce a broad spectrum of growth-inhibitory, proapoptotic, and antiangiogenic responses. Treatment of Panc1, Panc28, and L3.6pL pancreatic cancer cells with low micromolar concentrations of CDDO or CDDO-Me resulted in growth inhibition, induction of apoptosis, and down-regulation of cyclin D1, survivin, vascular endothelial growth factor (VEGF), and its receptor (VEGFR2). RNA interference studies indicate that these repressed genes are regulated by specificity protein (Sp) transcription factors Sp1, Sp3, and Sp4, and Western blot analysis of lysates from pancreatic cancer cells treated with CDDO and CDDO-Me shows for the first time that both compounds decreased the expression of Sp1, Sp3, and Sp4. Moreover, CDDO-Me (7.5 mg/kg/day) also inhibited pancreatic human L3.6pL tumor growth and down-regulated Sp1, Sp3, and Sp4 in tumors using an orthotopic pancreatic cancer model. CDDO-Me also induced reactive oxygen species (ROS) and decreased mitochondrial membrane potential (MMP) in Panc1 and L3.6pL cells, and cotreatment with antioxidants (glutathione and dithiothreitol) blocked the formation of ROS, reversed the loss of MMP, and inhibited down-regulation of Sp1, Sp3, and Sp4. Repression of Sp and Sp-dependent genes by CDDO-Me was due to the down-regulation of microRNA-27a and induction of zinc finger and BTB domain containing 10 (ZBTB10), an Sp repressor, and these responses were also reversed by antioxidants. Thus, the anticancer activity of CDDO-Me is due, in part, to activation of ROS, which in turn targets the microRNA-27a:ZBTB10-Sp transcription factor axis. This results in decreased expression of Sp-regulated genes, growth inhibition, induction of apoptosis, and antiangiogenic responses.

背景与目标： ：抗癌剂2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid（CDDO）及其甲酯（CDDO-Me）通常诱导广谱的生长抑制，促凋亡和抗血管生成回应。用低摩尔浓度的CDDO或CDDO-Me处理Panc1，Panc28和L3.6pL胰腺癌细胞会导致生长抑制，细胞凋亡诱导以及下调cyclin D1，survivin，血管内皮生长因子（VEGF），及其受体（VEGFR2）。 RNA干扰研究表明，这些阻抑的基因受特异性蛋白（Sp）转录因子Sp1，Sp3和Sp4调控，并且用CDDO和CDDO-Me处理的胰腺癌细胞裂解物的蛋白质印迹分析首次显示这两种化合物降低了Sp1，Sp3和Sp4的表达。此外，使用原位胰腺癌模型，CDDO-Me（7.5 mg / kg /天）还抑制了胰腺人类L3.6pL肿瘤的生长，并下调了肿瘤中Sp1，Sp3和Sp4的表达。 CDDO-Me还可以诱导Panc1和L3.6pL细胞中的活性氧（ROS）并降低线粒体膜电位（MMP），并且与抗氧化剂（谷胱甘肽和二硫苏糖醇）共同处理可阻止ROS的形成，逆转MMP的丧失并受到抑制下调Sp1，Sp3和Sp4。 CDDO-Me抑制Sp和Sp依赖性基因是由于microRNA-27a的下调以及诱导含有Sp阻遏物的10个锌指和BTB结构域（ZBTB10）的诱导，并且这些反应也被抗氧化剂逆转。因此，CDDO-Me的抗癌活性部分归因于ROS的激活，后者又靶向microRNA-27a：ZBTB10-Sp转录因子轴。这导致Sp调节基因的表达降低，生长抑制，凋亡诱导和抗血管生成反应。

BACKGROUND & AIMS: :Treatment of ErbB2-overexpressing BT474 and MDA-MB-453 breast cancer cells with 1 to 10 μmol/L betulinic acid inhibited cell growth, induced apoptosis, downregulated specificity protein (Sp) transcription factors Sp1, Sp3, and Sp4, and decreased expression of ErbB2. Individual or combined knockdown of Sp1, Sp3, Sp4 by RNA interference also decreased expression of ErbB2 and this response was because of repression of YY1, an Sp-regulated gene. Betulinic acid-dependent repression of Sp1, Sp3, Sp4, and Sp-regulated genes was due, in part, to induction of the Sp repressor ZBTB10 and downregulation of microRNA-27a (miR-27a), which constitutively inhibits ZBTB10 expression, and we show for the first time that the effects of betulinic acid on the miR-27a:ZBTB10-Sp transcription factor axis were cannabinoid 1 (CB1) and CB2 receptor-dependent, thus identifying a new cellular target for this anticancer agent.

背景与目标： ：用1至10μmol/ L的桦木酸处理过表达ErbB2的BT474和MDA-MB-453乳腺癌细胞抑制细胞生长，诱导细胞凋亡，下调特异性蛋白（Sp）转录因子Sp1，Sp3和Sp4并降低表达的ErbB2。 RNA干扰对Sp1，Sp3，Sp4的单独或组合敲低也降低了ErbB2的表达，这种反应是由于抑制了Sp调控的基因YY1。苯丙酸对Sp1，Sp3，Sp4和Sp调控基因的抑制部分归因于Sp阻遏物ZBTB10的诱导和microRNA-27a（miR-27a）的下调，后者构成性地抑制了ZBTB10的表达，我们首次表明，桦木酸对miR-27a：ZBTB10-Sp转录因子轴的影响是大麻素1（CB1）和CB2受体依赖性的，从而确定了该抗癌剂的新细胞靶标。

BACKGROUND & AIMS: :Celastrol (CSL) is a naturally occurring triterpenoid acid that exhibits anticancer activity, and in KU7 and 253JB-V bladder cells, CSL induced apoptosis, inhibited growth, colony formation and migration and CSL decreased bladder tumor growth in vivo. CSL also decreased expression of specificity protein (Sp) transcription factors Sp1, Sp3 and Sp4 and several Sp-regulated genes/proteins including vascular endothelial growth factor, survivin and cyclin D1 and fibroblast growth factor receptor-3, a potential drug target for bladder cancer therapy, has now been characterized as an Sp-regulated gene downregulated by CSL. The mechanism of Sp downregulation by CSL was cell context-dependent due to activation of proteosome-dependent (KU7) and -independent (253JB-V) pathways. In 253JB-V cells, CSL induced reactive oxygen species (ROS) and inhibitors of ROS blocked CSL-induced growth inhibition and repression of Sp1, Sp3 and Sp4. This response was due to induction of the Sp repressors ZBTB10 and ZBTB4 and downregulation of miR-27a and miR-20a/17-5p, respectively, which regulate expression of these transcriptional repressors. Thus, the anticancer activity of CSL in 253JB-V cells is due to induction of ROS and ROS-mediated induction of Sp repressors (ZBTB4/ZBTB10) through downregulation of miR-27a and miR-20a/17-5p.

背景与目标： ：Celastrol（CSL）是天然存在的三萜酸，具有抗癌活性，在KU7和253JB-V膀胱细胞中，CSL诱导凋亡，抑制生长，集落形成和迁移，并降低体内膀胱肿瘤的生长。 CSL还降低了特异性蛋白（Sp）转录因子Sp1，Sp3和Sp4以及一些Sp调控的基因/蛋白的表达，包括血管内皮生长因子，survivin和cyclin D1和成纤维细胞生长因子受体3，这是膀胱癌的潜在药物靶标现在，这种疗法的特征是被CSL下调了Sp调控的基因。 CSL对Sp的下调机制是由于蛋白体依赖性（KU7）和非依赖性（253JB-V）途径的激活而与细胞环境有关。在253JB-V细胞中，CSL诱导了活性氧（ROS），ROS抑制剂阻止了CSL诱导的Sp1，Sp3和Sp4的生长抑制和抑制。该应答是由于分别诱导Sp阻遏物ZBTB10和ZBTB4的诱导以及miR-27a和miR-20a / 17-5p的下调，它们分别调节这些转录阻遏物的表达。因此，CSL在253JB-V细胞中的抗癌活性是由于通过下调miR-27a和miR-20a / 17-5p诱导了ROS和ROS介导的Sp阻遏物（ZBTB4 / ZBTB10）的诱导。

BACKGROUND & AIMS: BACKGROUND:Betulinic acid (BA) inhibits growth of several cancer cell lines and tumors and the effects of BA have been attributed to its mitochondriotoxicity and inhibition of multiple pro-oncogenic factors. Previous studies show that BA induces proteasome-dependent degradation of specificity protein (Sp) transcription factors Sp1, Sp3 and Sp4 in prostate cancer cells and this study focused on the mechanism of action of BA in colon cancer cells. METHODS:The effects of BA on colon cancer cell proliferation and apoptosis and tumor growth in vivo were determined using standardized assays. The effects of BA on Sp proteins and Sp-regulated gene products were analyzed by western blots, and real time PCR was used to determine microRNA-27a (miR-27a) and ZBTB10 mRNA expression. RESULTS:BA inhibited growth and induced apoptosis in RKO and SW480 colon cancer cells and inhibited tumor growth in athymic nude mice bearing RKO cells as xenograft. BA also decreased expression of Sp1, Sp3 and Sp4 transcription factors which are overexpressed in colon cancer cells and decreased levels of several Sp-regulated genes including survivin, vascular endothelial growth factor, p65 sub-unit of NFκB, epidermal growth factor receptor, cyclin D1, and pituitary tumor transforming gene-1. The mechanism of action of BA was dependent on cell context, since BA induced proteasome-dependent and proteasome-independent downregulation of Sp1, Sp3 and Sp4 in SW480 and RKO cells, respectively. In RKO cells, the mechanism of BA-induced repression of Sp1, Sp3 and Sp4 was due to induction of reactive oxygen species (ROS), ROS-mediated repression of microRNA-27a, and induction of the Sp repressor gene ZBTB10. CONCLUSIONS:These results suggest that the anticancer activity of BA in colon cancer cells is due, in part, to downregulation of Sp1, Sp3 and Sp4 transcription factors; however, the mechanism of this response is cell context-dependent.

背景与目标： 背景：桦木酸（BA）抑制几种癌细胞系和肿瘤的生长，BA的作用归因于其线粒体毒性和多种促癌因子的抑制作用。先前的研究表明，BA可以诱导前列腺癌细胞中蛋白酶体依赖的特异性蛋白（Sp）转录因子Sp1，Sp3和Sp4降解，而这项研究的重点是BA在结肠癌细胞中的作用机理。
方法：采用标准化方法测定BA对体内结肠癌细胞增殖，凋亡和肿瘤生长的影响。通过蛋白质印迹分析BA对Sp蛋白和Sp调控基因产物的影响，并使用实时荧光定量PCR测定microRNA-27a（miR-27a）和ZBTB10 mRNA的表达。
结果：BA抑制RKO和SW480结肠癌细胞的生长并诱导其凋亡，并抑制携带RKO细胞作为异种移植物的无胸腺裸鼠的肿瘤生长。 BA还降低了在结肠癌细胞中过表达的Sp1，Sp3和Sp4转录因子的表达，并降低了几个Sp调节基因的水平，包括Survivin，血管内皮生长因子，NFκB的p65亚基，表皮生长因子受体，细胞周期蛋白D1和垂体肿瘤转化基因-1。 BA的作用机制取决于细胞的情况，因为BA分别在SW480和RKO细胞中分别诱导了蛋白酶体依赖性和蛋白酶体依赖性Sp1，Sp3和Sp4的下调。在RKO细胞中，BA诱导Sp1，Sp3和Sp4抑制的机制是由于活性氧（ROS）的诱导，ROS介导的microRNA-27a的抑制以及Sp阻遏物基因ZBTB10的诱导。
结论：这些结果表明BA在结肠癌细胞中的抗癌活性部分归因于Sp1，Sp3和Sp4转录因子的下调。但是，此响应的机制取决于单元上下文。

BACKGROUND & AIMS: :Genistein, an isoflavone isolated from soybean, has been found to be a potent antitumor agent. However, both the effect of genistein on human uveal melanoma cells and the precise mechanism by which genistein suppresses tumorigenesis remain unclear. In the present study, we explored the possible activity of genistein to inhibit human uveal melanoma cell growth and investigated the possible role of genistein on microRNA-27a (miR-27a) as well as its target gene zinc finger and BTB domain containing 10 (ZBTB10) expression levels. The results suggested a significant inhibition of uveal melanoma cell growth in a time- and dose-related manner. In vivo study also indicated treatment groups with genistein could significantly inhibit the growth of xenografts. Further functional assays revealed that the levels of miR-27a and its target gene ZBTB10 were significantly different based on the dose of genistein. In conclusion, the present study demonstrates that genistein exerts growth inhibitory activities in human uveal melanoma cells. Moreover, we for the first time report a correlation between antitumor activity of genistein and miR-27a mediated regulatory mechanism.

背景与目标： ：Genistein是一种从大豆中分离出来的异黄酮，已被发现是一种有效的抗肿瘤药。然而，金雀异黄素对人葡萄膜黑色素瘤细胞的作用以及金雀异黄素抑制肿瘤发生的确切机制尚不清楚。在本研究中，我们探讨了染料木黄酮抑制人葡萄膜黑色素瘤细胞生长的可能活性，并研究了染料木黄酮对microRNA-27a（miR-27a）及其靶基因锌指和含10个BTB结构域（ZBTB10）的可能作用）的表达水平。结果表明以时间和剂量相关的方式显着抑制葡萄膜黑色素瘤细胞的生长。体内研究还表明，染料木黄酮治疗组可以显着抑制异种移植物的生长。进一步的功能分析表明，基于染料木黄酮的剂量，miR-27a及其靶基因ZBTB10的水平显着不同。总之，本研究证明金雀异黄素在人葡萄膜黑色素瘤细胞中具有生长抑制活性。此外，我们首次报道了染料木黄酮的抗肿瘤活性与miR-27a介导的调节机制之间的相关性。

BACKGROUND & AIMS: :Triterpenoids such as betulinic acid (BA) and synthetic analogs of oleanolic acid [2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO)] and glycyrrhetinic acid [2-cyano-3,11-dioxo-18β-oleana-1,12-dien-30-oc acid (CDODA)] are potent anticancer agents that exhibit antiproliferative, antiangiogenic, anti-inflammatory and pro-apoptotic activities. Although their effects on multiple pathways have been reported, unifying mechanisms of action have not been reported. Studies in this laboratory have now demonstrated that several triterpenoids including BA and some derivatives, celastrol, methyl ursolate, β-boswellic acid derivatives, and the synthetic analogs CDDO, CDODA and their esters decreased expression of specificity protein (Sp) transcription factors and several pro-oncogenic Sp-regulated genes in multiple cancer cell lines. The mechanisms of this response are both compound- and cell context-dependent and include activation of both proteasome-dependent and -independent pathways. Triterpenoid-mediated induction of reactive oxygen species (ROS) has now been characterized as an important proteasome-independent pathway for downregulation of Sp transcription factors. ROS decreases expression of microRNA-27a (miR-27a) and miR-20a/miR-17-5p and this results in the induction of the transcriptional "Sp-repressors" ZBTB10 and ZBTB4, respectively, which in turn downregulate Sp and Sp-regulated genes. Triterpenoids also activate or deactive nuclear receptors and G-protein coupled receptors, and these pathways contribute to their antitumorigenic activity and may also play a role in targeting Sp1, Sp3 and Sp4 which are highly overexpressed in multiple cancers and appear to be important for maintaining the cancer phenotype.

背景与目标： ：三萜类化合物，例如桦木酸（BA）和齐墩果酸[2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid（CDDO）]和甘草次酸[2-cyano-3， 11-二氧代-18β-oleana-1,12-dien-30-oc酸（CDODA）]是有效的抗癌剂，具有抗增殖，抗血管生成，抗炎和促凋亡的活性。尽管已经报道了它们对多种途径的作用，但尚未报道统一的作用机理。现在，该实验室的研究表明，三萜类化合物包括BA和某些衍生物，天青素，尿酸甲酯，β-乳香酸衍生物，以及合成类似物CDDO，CDODA及其酯可降低特异性蛋白（Sp）转录因子的表达，并且一些多种癌细胞系中致癌的Sp调控基因。这种反应的机制既取决于化合物又取决于细胞，并且包括蛋白酶体依赖性和非依赖性途径的激活。三萜类化合物介导的活性氧（ROS）的诱导现已被表征为下调Sp转录因子的重要的蛋白酶体独立途径。 ROS降低了microRNA-27a（miR-27a）和miR-20a / miR-17-5p的表达，这分别导致了转录“ Sp阻遏物” ZBTB10和ZBTB4的诱导，进而下调了Sp和Sp-调控基因。三萜类化合物还激活或失活的核受体和G蛋白偶联受体，这些通路有助于其抗肿瘤发生活性，并且还可能在靶向Sp1，Sp3和Sp4方面发挥作用，而Sp1，Sp3和Sp4在多种癌症中过度表达，对于维持肿瘤的发生很重要。癌症表型。

BACKGROUND & AIMS: BACKGROUND:Genome-wide association studies (GWASs) have identified various genes associated with asthma, yet, causal genes or single nucleotide polymorphisms (SNPs) remain elusive. We sought to dissect functional genes/SNPs for asthma by combining expression quantitative trait loci (eQTLs) and GWASs. METHODS:Cis-eQTL analyses of 34 asthma genes were performed in cells from human bronchial epithelial biopsy (BEC, n = 107) and from bronchial alveolar lavage (BAL, n = 94). RESULTS:For TSLP-WDR36 region, rs3806932 (G allele protective against eosinophilic esophagitis) and rs2416257 (A allele associated with lower eosinophil counts and protective against asthma) were correlated with decreased expression of TSLP in BAL (P = 7.9 × 10(-11) and 5.4 × 10(-4) , respectively) and BEC, but not WDR36. Surprisingly, rs1837253 (consistently associated with asthma) showed no correlation with TSLP expression levels. For ORMDL3-GSDMB region, rs8067378 (G allele protective against asthma) was correlated with decreased expression of GSDMB in BEC and BAL (P = 1.3 × 10(-4) and 0.04) but not ORMDL3. rs992969 in the promoter region of IL33 (A allele associated with higher eosinophil counts and risk for asthma) was correlated with increased expression of IL33 in BEC (P = 1.3 × 10(-6) ) but not in BAL. CONCLUSIONS:Our study illustrates cell-type-specific regulation of the expression of asthma-related genes documenting SNPs in TSLP, GSDMB, IL33, HLA-DQB1, C11orf30, DEXI, CDHR3, and ZBTB10 affect asthma risk through cis-regulation of its gene expression. Whenever possible, disease-relevant tissues should be used for transcription analysis. SNPs in TSLP may affect asthma risk through up-regulating TSLP mRNA expression or protein secretion. Further functional studies are warranted.

背景与目标： 背景：全基因组关联研究（GWAS）已鉴定出与哮喘相关的各种基因，但因果基因或单核苷酸多态性（SNP）仍然难以捉摸。我们试图通过结合表达定量性状基因座（eQTL）和GWASs来分析哮喘的功能基因/ SNP。
方法：对来自人支气管上皮活检（BEC，n = 107）和支气管肺泡灌洗（BAL，n = 94）的34种哮喘基因进行Cis-eQTL分析。
结果：对于TSLP-WDR36区域，rs3806932（G等位基因对嗜酸性粒细胞性食管炎有保护作用）和rs2416257（等位基因与嗜酸性粒细胞减少和哮喘相关）与BAL中TSLP的表达降低相关（P = 7.9×10（-11） ）和5.4×10（-4））和BEC，但不包括WDR36。出乎意料的是，rs1837253（与哮喘相关）始终与TSLP表达水平无关。对于ORMDL3-GSDMB区，rs8067378（对哮喘具有保护作用的G等位基因）与BEC和BAL中GSDMB的表达降低相关（P = 1.3×10（-4）和0.04），而与ORMDL3无关。 IL33（与较高的嗜酸性粒细胞计数和哮喘风险有关的等位基因）启动子区域中的rs992969与BEC中IL33表达的增加相关（P = 1.3×10（-6）），但与BAL中的IL33表达无关。
结论：我们的研究阐明了哮喘相关基因表达的细胞类型特异性调控，该基因表达TSLP，GSDMB，IL33，HLA-DQB1，C11orf30，DEXI，CDHR3和ZBTB10中的SNPs，通过顺式调控其基因影响哮喘风险表达。只要有可能，应使用与疾病相关的组织进行转录分析。 TSLP中的SNP可能通过上调TSLP mRNA表达或蛋白质分泌来影响哮喘风险。有必要进行进一步的功能研究。

BACKGROUND & AIMS: :Methyl 2-trifluoromethyl-3,11-dioxo-18β-olean-1,12-dien-3-oate (CF3DODA-Me) is derived synthetically from glycyrrhetinic acid, a major component of licorice, and this compound induced reactive oxygen species (ROS) in RD and Rh30 rhabdomyosarcoma (RMS) cells. CF3DODA-Me also inhibited growth and invasion and induced apoptosis in RMS cells, and these responses were attenuated after cotreatment with the antioxidant glutathione, demonstrating the effective anticancer activity of ROS in RMS. CF3DODA-Me also downregulated expression of specificity protein (Sp) transcription factors Sp1, Sp3, and Sp4 and prooncogenic Sp-regulated genes including PAX3-FOXO1 (in Rh30 cells). The mechanism of CF3DODA-Me-induced Sp-downregulation involved ROS-dependent repression of c-Myc and cMyc-regulated miR-27a and miR-17/20a, and this resulted in induction of the miRNA-regulated Sp repressors ZBTB4, ZBTB10, and ZBTB34. The cell and tumor growth effects of CF3DODA-Me further emphasize the sensitivity of RMS cells to ROS inducers and their potential clinical applications for treating this deadly disease. IMPLICATIONS: CF3DODA-Me and HDAC inhibitors that induce ROS-dependent Sp downregulation could be developed for clinical applications in treating rhabdomyosarcoma.

背景与目标： ：2-三氟甲基-3,11-二氧杂-18β-油酸酯-1,12-二烯-3-酸酯（CF3DODA-Me）合成自甘草酸的主要成分甘草次酸，该化合物可诱导活性氧（ROS）在RD和Rh30横纹肌肉瘤（RMS）细胞中。 CF3DODA-Me还抑制RMS细胞的生长和侵袭并诱导细胞凋亡，在与抗氧化剂谷胱甘肽共同处理后，这些反应减弱，证明ROS在RMS中具有有效的抗癌活性。 CF3DODA-Me还下调了特异性蛋白（Sp）转录因子Sp1，Sp3和Sp4的表达以及包括PAX3-FOXO1在内的促癌性Sp调控的基因（在Rh30细胞中）。 CF3DODA-Me诱导的Sp下调的机制涉及ROS依赖的c-Myc和cMyc调节的miR-27a和miR-17 / 20a的阻遏作用，这导致了miRNA调节的Sp阻遏物ZBTB4，ZBTB10，和ZBTB34。 CF3DODA-Me对细胞和肿瘤的生长作用进一步强调了RMS细胞对ROS诱导剂的敏感性及其在治疗这种致命疾病中的潜在临床应用。结论：可以诱导ROS依赖性Sp下调的CF3DODA-Me和HDAC抑制剂可用于治疗横纹肌肉瘤的临床应用。