关键词： 食管鳞状细胞癌   无孢蛋白   癌旁正常组织   癌基因  

摘要： 目的探讨无孢蛋白(Asporin)在食管鳞状细胞癌发生发展中的作用及临床意义。方法收集2019年12月⁃2021年11月选取涟水人民医院收治的140例食管鳞状细胞癌患者。随机选取60例新鲜切取的食管鳞状细胞癌(ESCC)组织及癌旁正常组织,利用Western blot、RT⁃PCR技术检测Asporin的蛋白及mRNA表达水平;80例ESCC组织及其相应癌旁组织的石蜡标本,利用免疫组化技术进一步检测Asporin的表达情况;利用卡方检验、Kaplan⁃Meier分析以及COX回归分析验证Asporin的表达与ESCC患者临床病理特征及预后的关系;Transwell侵袭实验研究Asporin沉默后对ESCC细胞株侵袭转移功能的影响;Pearson相关分析评价Asporin与基质金属蛋白酶2(MMP2)之间的相关性。结果Western blot和RT⁃PCR结果显示ESCC的Asporin蛋白和mRNA表达水平明显高于其癌旁正常组织(P<0.05);免疫组化结果显示,Asporin在ESCC组织中的表达明显高于癌旁组织,且Asporin在ESCC的高表达率为65%(52/80);Asporin表达水平与患者的淋巴结转移情况及TNM分期有显著相关性(P<0.05),与其他临床病理特征无显著相关性(P>0.05);Kaplan⁃Meier分析结果表明,高表达Asporin的ESCC患者无病生存期短于低表达ASPN的患者(P<0.01);COX回归分析显示,低表达Asporin的ESCC患者死亡风险低于高表达ASPN的患者,HR=0.106(95%CI:0.024⁃0.469),Asporin是影响ESCC患者预后的独立危险因素(P=0.003);Transwell侵袭实验表明,经siRNA沉默Asporin后,与阴性对照组相比,Asporin敲低组的侵袭能力明显降低;Pearson相关分析显示,Asporin的表达水平与MMP2表达存在中度正相关关系(r=0.61,P<0.01),MMP2在恶性肿瘤侵袭转移过程中起关键作用。结论Asporin可能作为一种癌基因,参与食管鳞状细胞癌的发生发展。

关键词： Asporin   Osteocalcin   Osteopontin   Osteoblast   BMP2   Runx2   RNA-Seq   Anti-miR   Fetuin A   Differentiation  

摘要： BackgroundCalcific aortic valve disease (CAVD) is the second leading cause of adult heart diseases. The purpose of this study is to investigate whether miR-101-3p plays a role in the human aortic valve interstitial cells (HAVICs) calcification and the underlying *** RNA deep sequencing and qPCR analysis were used to determine changes in microRNA expression in calcified human aortic *** data showed that miR-101-3p levels were increased in the calcified human aortic valves. Using cultured primary HAVICs, we demonstrated that the miR-101-3p mimic promoted calcification and upregulated the osteogenesis pathway, while anti-miR-101-3p inhibited osteogenic differentiation and prevented calcification in HAVICs treated with the osteogenic conditioned medium. Mechanistically, miR-101-3p directly targeted cadherin-11 (CDH11) and Sry-related high-mobility-group box 9 (SOX9), key factors in the regulation of chondrogenesis and osteogenesis. Both CDH11 and SOX9 expressions were downregulated in the calcified human HAVICs. Inhibition of miR-101-3p restored expression of CDH11, SOX9 and ASPN and prevented osteogenesis in HAVICs under the calcific ***-101-3p plays an important role in HAVIC calcification through regulation of CDH11/SOX9 expression. The finding is important as it reveals that miR-1013p may be a potential therapeutic target for calcific aortic valve disease.

关键词： alveolar bone   fiber disorder   Flexcell FX-5000 compression system   occlusal hypofunction   overeruption   periodontal ligament   PLAP-1   asporin  

摘要： Background and ObjectiveTeeth overeruption is a problem of clinical significance, but the underlying mechanism how changes in external occlusal force convert to the periodontium remodeling signals has been a largely under explored domain. And recently, periodontal ligament-associated protein-1 (PLAP-1)/asporin was found to play a pivotal role in maintaining periodontal homeostasis. The aim of this study was to explore the function of PLAP-1 in the periodontally hypofunctional tissue turnover. MethodsAfter extracting left maxillary molars in mice, the left and right mandibular molars were distributed into hypofunction group (HG) and control group (CG), respectively. Mice were sacrificed for radiographic, histological, and molecular biological analyses after 1, 4 and 12 weeks. In vitro, dynamic compression was applied using Flexcell FX-5000 Compression System to simulate intermittent occlusal force. The expression of PLAP1 in loaded and unloaded human periodontal ligament cells (hPDLCs) was compared, and its molecular biological effects were further explored by small interfering RNA (siRNA) targeting PLAP1. ResultsIn vivo, fiber disorder in periodontal ligament (PDL), bone apposition at furcation regions, and bone resorption in alveolar bone were illustrated in the HG compared with the CG. In addition, PLAP-1 positive area decreased significantly in PDL following occlusal unloading. In vitro, the loss of compressive loading relatively downregulated PLAP1 expression, which was essential to promote collagen I but inhibit osterix and osteocalcin expression in hPDLCs. ConclusionsPLAP-1 presumably plays a pivotal role in occlusal force-regulated periodontal homeostasis by facilitating collagen fiber synthesis in hPDLCs and suppressing excessive osteoblast differentiation, further preventing teeth from overeruption. Further evidence in PLAP-1 conditional knockout mice is needed.

关键词： 神经内分泌肿瘤   甲状腺髓样癌   恶性增殖   细胞侵袭   遗传特性   甲状腺癌   MTC   放化疗  

摘要： 甲状腺髓样癌(medullary thyroid cancer, MTC)是一种恶性的神经内分泌肿瘤,由甲状腺C细胞恶性增殖产生,约占所有甲状腺癌的2%~3%。根据疾病的遗传特性,MTC可分为遗传性和散发性两大类。尽管局限性不伴有远处转移的散发性MTC具有良好的预后,10年生存率能够达到70%~80%,但是MTC细胞侵袭性高,很多患者初诊时已是晚期,传统的“手术+放化疗模式”获益有限,10年生存率降到仅有10%。

关键词： Asporin   ECM   Cardiac remodeling   Fibrosis   Ischemia  

摘要： Heart failure is accompanied by adverse cardiac remodeling involving extracellular matrix (ECM). Cardiac ECM acts as a major reservoir for many proteins including growth factors, cytokines, collagens, and proteoglycans. Activated fibroblasts during cardiac injury can alter the composition and activity of these ECM proteins. Through unbiased analysis of a microarray dataset of human heart tissue comparing normal hearts (n = 135) to hearts with ischemic cardiomyopathy (n = 94), we identified Asporin (ASPN) as the top differentially regulated gene (DEG) in ischemic cardiomyopathy;its gene-ontology terms relate closely to fibrosis and cell death. ASPN is a Class I small leucine repeat protein member implicated in cancer, osteoarthritis, and periodontal ligament mineralization. However, its role in cardiac remodeling is still unknown. Here, we initially confirmed our big dataset analysis through cells, mice, and clinical atrial biopsy samples to demonstrate increased Aspn expression after pressure overload or cardiac ischemia/reperfusion injury. We tested the hypothesis that Aspn, being a TGFb1 inhibitor, can attenuate fibrosis in mouse models of cardiac injury. We found that Aspn is released by cardiac fibroblasts and attenuates TGFb signaling. Moreover, Aspn(-/-) mice displayed increased fibrosis and decreased cardiac function after pressure overload by transverse aortic constriction (TAC) in mice. In addition, Aspn protected cardiomyocytes from hypoxia/reoxygenation-induced cell death and regulated mitochondrial bioenergetics in cardiomyocytes. Increased infarct size after ischemia/reperfusion injury in Aspn(-/-) mice confirmed Aspn's contribution to cardiomyocyte viability. Echocardiography revealed greater reduction in left ventricular systolic function post-I/R in the Aspn(-/-) animals compared to wild type. Furthermore, we developed an ASPN-mimic peptide using molecular modeling and docking which when administered to mice prevented TAC-induced fibrosis and preserv

关键词： 椎间盘退变   纤维环修复   生物疗法   组织工程   基因治疗  

摘要： 纤维环损伤与椎间盘退行性疾病(IVDD)的发病密切相关,也是导致椎间盘术后疗效不佳的重要因素,恢复纤维环结构和功能的完整性对缓解甚至逆转IVDD有重要意义。传统的椎间盘纤维环修复策略仅完成物理修复而不能恢复纤维环的生物力学特性,并不是理想的修复方式。近年来,应用生物学方法来修复纤维环取得了一些进展。本文综述了目前对椎间盘纤维环损伤的认识,并介绍了包括细胞移植、纤维环组织工程以及基因治疗在内的各种生物疗法在椎间盘纤维环再生策略方面的最新进展,以期为未来的研究方向选择提供帮助。

摘要： V-241

关键词： Asporin   CAF   CAF-educated fibroblasts   CEF   macrophages   MC-CAF   mesothelial cells  

摘要： Tumors contain various stromal cells that support cancer progression. Some types of cancer, such as scirrhous gastric cancer, are characterized by large areas of fibrosis accompanied by cancer-associated fibroblasts (CAFs). Asporin (ASPN) is a small leucine-rich proteoglycan highly expressed in CAFs of various tumors. ASPN accelerates CAF migration and invasion, resulting in CAF-led cancer cell invasion. In addition, ASPN further upregulated the expression of genes specific to a characteristic subgroup of fibroblasts in tumors. These cells were preferentially located at the tumor periphery and could be generated by a unique mechanism involving the CAF-mediated education of normal fibroblasts (CEFs). In this review, we at first describe recent findings regarding the function of ASPN in the tumor microenvironment, as well as the mechanism involved in the generation of CEFs. CAFs are derived from heterogeneous origins besides resident normal fibroblasts. Among them, CAFs derived from mesothelial cells (mesothelial cell-derived CAF [MC-CAFs]) play pivotal roles in peritoneal carcinomatosis. We observed that MC-CAFs on the surfaces of organs also participate in tumor formation by infiltrating into the parenchyma, promoting local invasion by gastric cancers. This review also highlights the potential functions of macrophages in the formation of MC-CAFs in gastric cancers, by transfer the contents of cancer cell-derived extracellular vesicles.

关键词： Osteoblasts   Epigenetics   Transcription factors   Cell differentiation   Bone regeneration  

摘要： Periodontal dental ligament (PDL) is composed of heterogeneous population of mesenchymal progenitor cells. The mechanisms that regulate the differentiation of these cells towards osteoblast/cementoblast phenotype are not fully understood. Some studies have demonstrated that is possible to change the pattern of cell differentiation via epigenetic mechanisms. The proposal of this study was to investigate whether 5-aza-2'-deoxycytidine (5-azadC) treatment would stimulate the osteoblast/cementoblast differentiation of periodontal ligament mesenchymal progenitor cells (PDL-CD105+ enriched cells), characterized as low osteoblast potential, through bone morphogenetic protein-2 (BMP-2) modulation. PDL-CD105+ cells from a single donor were cloned and characterized in two populations as high osteoblast/cementoblast potential (HOP) and low osteoblast/cementoblast potential (LOP) by mineralization in vitro and expression of osteogenic gene markers, such as runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), bone morphogenetic protein 2 (BMP-2) and asporin (ASPN). Next, two LOP clones (L1 and L2) were pretreated with 5-aza-dC (10 mu M) for 48 h, cultured under osteogenic condition and evaluated for mineralized matrix in vitro, transcription modulation of osteogenic gene markers, methylated and hydroxymethylated DNA levels of BMP-2 and ASPN and intracellular/ extracellular expression of BMP-2 protein. LOP clones showed high expression of ASPN transcripts associated with low mRNA levels of BMP-2, RUNX2, ALP, and OCN. 5-aza-dC treatment raised hydroxymethylated DNA levels of BMP-2 and increased the expression of BMP-2 transcripts in both LOP clones. However, BMP-2 protein (intracellular and secreted forms) was detected only in L1 cell clones, in which it was observed an increased expression of osteoblast/cementoblast markers (RUNX2, ALP, OCN) associated with higher mineralization in vitro. In L2 cell clones, 5-aza-dC increased gene expression

关键词： asporin   CAF-educated fibroblasts   CD8(+) T cells   IGF-I   kynurenine  

摘要： In some tumors, a small number of cancer cells are scattered in a large fibrotic stroma. Here, we demonstrate a novel mechanism for expansion of pro-tumor fibroblasts via cancer-associated fibroblast (CAF)-mediated education of normal fibroblasts (NFs). When NFs were incubated with conditioned medium from CAFs, the resulting CAF-educated fibroblasts (CEFs) generated reactive oxygen species, which induced NF-kappa B-mediated expression of inflammatory cytokines and the extracellular matrix protein asporin (ASPN), while expression of a common CAF marker gene, alpha-SMA, was not increased. ASPN further increased CEF expression of downstream molecules, including indoleamine 2,3-dioxygenase 1 (IDO-1), kynureninase (KYNU), and pregnancy-associated plasma protein-A (PAPP-A). These CEFs induce cytocidal effects against CD8(+) T cells and IGF-I activation in cancer cells. CEFs were generated without cancer cells by the direct mixture of NFs and CAFs in mouse xenografts, and once CEFs were generated, they sequentially educated NFs, leading to continuous generation of CEFs. In diffuse-type gastric cancers, ASPN(high)/IDO-1(high)/KYNUhigh/alpha-SMA(-) CEFs were located at the distal invading front. These CEFs expanded in the fibrotic stroma and caused dissemination of cancer cells. ASPN may therefore be a key molecule in facilitating tumor spreading and T-cell suppression.