圣草素促进成骨细胞自噬减轻糖皮质激素诱导的细胞凋亡

背景：糖皮质激素诱导的骨质疏松症是系统性糖皮质激素治疗的常见并发症,主要表现为对成骨细胞的抑制作用。圣草素可抑制破骨细胞分化和卵巢切除术引起的骨质疏松,然而,圣草素是否调节糖皮质激素诱导的成骨细胞凋亡尚不清楚。目的：探究圣草素是否在地塞米松诱导的成骨细胞凋亡中发挥作用,及其潜在的作用机制。方法：采用不同浓度(0,0.5,1,2.5,5,10μmol/L)的圣草素或自噬抑制剂3-甲基腺嘌呤(5μmol/L)处理地塞米松预处理的成骨细胞MC3T3-E1,然后用血红素加氧酶1的过表达载体(pcDNA-HMOX1)和空载体(pcDNA vector)转染MC3T3-E1细胞。分别采用CCK-8和流式细胞术检测细胞活力和凋亡;采用ELISA检测半胱氨酸天冬氨酸蛋白酶3活性;采用Western blotting检测自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ、p62、Atg5和Atg12表达,凋亡相关蛋白Bax和Bcl-2的表达,以及AMP蛋白激活激酶(AMPK)和p-AMPK蛋白表达。结果与结论：(1)低浓度的圣草素对MC3T3-E1细胞无毒性,且促进细胞增殖,并以剂量依赖性的方式促进自噬相关蛋白LC3-...     

长链非编码RNA H19与肿瘤关系的研究进展

长链非编码RNA(LncRNA)H19与肿瘤关系密切。H19在肺癌、乳腺癌、胃癌中起促癌作用,在肝癌中则具有双向作用。H19的作用机制有组织特异性,各不相同,多与微小RNA、影响原癌基因表达、EMT形成及甲基化状态下IGF2的印迹状态等有关,是一种有价值的潜在肿瘤生物靶标。     

长链非编码RNA H19在肝脏疾病中作用的研究进展

<正>人类基因组计划发现,仅有不到2%的基因组编码蛋白质,而近70%的基因组转录成非编码RNA^[1]。非编码RNA是指不具备蛋白质编码能力的RNA,包括转运RNA、核糖体RNA、小核仁RNA和长链非编码RNA(long noncoding RNA, lncRNA)。     

长链非编码RNA H19 靶向miR-141 调控卵巢癌细胞的迁移和侵袭行为

目的：探讨长链非编码RNA-H19靶向miR-141调控卵巢癌细胞的迁移和侵袭行为及其机制。方法：qPCR检测H19和miR-141在卵巢癌组织中的表达情况及H19在不同卵巢癌细胞株中的表达;分析H19和卵巢癌患者的临床病理资料之间的关联;双荧光素酶报告基因检测H19与miR-141之间的相互作用;划痕愈合实验和Transwell侵袭实验检测抑制H19后卵巢癌细胞的迁移和侵袭能力的变化情况,qPCR检测H19和miR-141之间的相互调控的关系;Western blot检测抑制H19后ZEB1蛋白的表达情况。结果：与正常卵巢组织相比,H19在卵巢癌组织中表达上调,miR-141在卵巢癌中的表达水平下调,与其他卵巢癌细胞株相比,ES-2细胞中H19表达最高;H19的表达与卵巢癌的病理分期相关以及淋巴结转移情况有关,随着分期越高,H19在卵巢癌组织中表达越高,淋巴结转移的患者中H19的表达也相对较高;双荧光素酶实验证实H19能与miR-141的3′ UTR特异性结合,可以调控miR-141的表达与活性;抑制H19的表达后可以抑制卵巢癌细胞的迁移和侵袭能力;miR-141可以负反馈调节H19的表达,抑制H19和miR-141的表达后,ZEB1蛋白的表达水平受到相应的抑制。结论：H19调控miR-141的表达通过影响ZEB1蛋白的表达参与卵巢癌细胞迁移和侵袭能力的调控。     

长链非编码RNA调控肿瘤细胞凋亡的研究进展

长链非编码RNA(long non-coding RNA,lncRNA)是一类长度>200个核苷酸、通常不编码蛋白质的RNA。近年研究表明,lncRNA在肿瘤的的发展过程中发挥抑癌或促癌作用,参与细胞增殖、凋亡等过程。本综述简要介绍lncRNA的生物学功能及其调控细胞凋亡的研究进展。     

长链非编码RNA调控椎间盘退变的研究进展

椎间盘退变(IDD)是导致腰痛的主要原因,目前在细胞和分子水平寻找IDD的机制得到了较多关注。已有研究报道长链非编码RNA(lncRNA)影响IDD的病理过程。本文综述了lncRNA对髓核细胞增殖、凋亡、衰老、自噬等方面的调控,总结了调控椎间盘内细胞增殖、凋亡和ECM合成与降解的lncRNA及相关靶基因或通路。值得注意的是,有的lncRNA不充当竞争内源性RNA(ceRNA),而是直接结合稳定相关微小RNA(miRNA),增强其miRNA表达从而调控IDD。此外,目前的研究主要报道的是lncRNA对椎间盘内髓核的调控,未来可进一步探究lncRNA对软骨终板和纤维环的调控,为lncRNA调控IDD提供更多的理论支持和依据。     

骨形态发生蛋白2诱导成骨分化与长链非编码RNA AK007000的影响

背景：长链非编码RNA调控一系列生理过程,被认为在发育、分化和代谢的基因调控中发挥重要的作用。MC3T3-E1、C2C12和C3H10T1/2细胞可向骨细胞、肌细胞等多个方向分化,用于肌肉骨骼等运动系统相关疾病的研究。 目的：观察长链非编码RNA在骨形态发生蛋白2诱导成骨分化中的作用。 方法：对MC3T3-E1、C2C12和C3H10T1/2细胞在骨形态发生蛋白2诱导下,成骨分化过程中的长链非编码RNA表达变化进行芯片分析,找到在3株细胞中同时变化的长链非编码RNA,siRNA干扰方法观察长链非编码RNA对骨形态发生蛋白2诱导成骨分化的影响,采用Real-Time PCR与碱性磷酸酶染色检测成骨相关指标。 结果与结论：骨形态发生蛋白2诱导MC3T3-E1、C2C12和C3H10T1/2成骨分化过程中,相应成骨指标增高,成肌指标肌细胞生成素降低。筛选出骨形态发生蛋白2诱导成骨分化过程中出发挥作用的长链非编码RNA AK007000。AK007000被干扰后成骨分化指标碱性磷酸酶、骨钙素、RUNX2、SP7表达下降,肌细胞生成素表达上升。因此,AK007000可能具有促进成骨抑制成肌作用。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

长链非编码RNA CRNDE调控胶质瘤细胞凋亡

目的 探讨长链非编码RNA CRNDE在胶质瘤细胞凋亡中发挥的功能和可能的作用机制.方法 将CRNDE基因干扰siRNA转染人胶质瘤U87细胞系,利用PE Annexin-V/7-AAD双染,流式细胞仪检测细胞凋亡情况,并利用Western方法检测细胞凋亡关键蛋白的表达变化.结果 siRNA干扰CRNDE表达的胶质瘤细胞中,细胞凋亡程度增加,凋亡相关蛋白Caspase3、Caspase7、Caspase9、PARP表达量明显升高.结论 CRNDE可能通过降低凋亡相关蛋白活性及表达水平,抑制胶质瘤细胞凋亡能力.     

长链非编码RNA调控巨噬细胞功能的作用机制研究进展

长链非编码RNA(lncRNA)是一类本身不编码蛋白、转录本长度超过200 bp的RNA。lncRNA最初被认为是RNA聚合酶Ⅱ转录的副产物,是一种“噪音”,不具有生物学功能。但近年研究证实,lncRNA参与X染色体沉默、染色体修饰和基因组修饰、转录激活、转录干扰、核内运输等过程,其调控作用被越来越多研究者关注。多个研究表明lncRNA在机体炎症反应过程中发挥重要作用。作为免疫细胞的巨噬细胞在机体免疫功能发挥和炎症反应过程中居主导地位,本文就lncRNA对巨噬细胞的调控作用及机制进行综述。     

Long non-coding RNA H19 regulates proliferation of ovarian granulosa cells via STAT3 in polycystic ovarian syndrome

IntroductionStudies have shown that long non-coding RNAs (lncRNA) are aberrantly expressed in polycystic ovarian syndrome (PCOS) ovaries and may have a role in PCOS development. In this study, the role and therapeutic implications of lncRNA H19 were investigated in PCOS ovaries and granulosa cells.Material and methodsqRT-PCR was used for expression analysis. Cell Counting Kit 8 (CCK-8) assay was used for cell viability and acridine orange/ethidium bromide (AO/EB) and Annexin V/propidium iodide staining was used to detect apoptosis. All transfections were carried out with Lipofectamine 2000 reagent. Western blot analysis was used for protein expression analysis.ResultsThe expression of lncRNA H19 was remarkably upregulated in the PCOS ovarian tissues as well as the granulosa cells. Suppression of lncRNA H19 expression caused the inhibition of KGN granulosa cell proliferation due to the triggering of apoptosis. Bioinformatic analysis revealed the presence of the GAS binding site for STAT3 in the promoter of lncRNA H19. Silencing of STAT3 suppressed the expression of lncRNA H19 in KGN cells and also halted their growth by triggering apoptosis. Co-transfect experiments revealed that STAT3 and lncRNA H19 silencing cause inhibition of KGN growth synergistically.ConclusionslncRNA H19 regulates the growth of ovarian granulosa cells and might prove to be a therapeutic target for management of PCOS.     

Ectopic expressed long non-coding RNA H19 contributes to malignant cell behavior of ovarian cancer

Recent studies have highlighted the role of long non-coding RNAs (lncRNAs) in carcinogenesis and have suggested that genes of this class might be used as biomarkers in cancer. However, whether lncRNAs are involved in ovarian cancer (OC) remains largely unknown. In the present study, we focused on lncRNAH19 and investigated the expression and functional role of H19 in OC. H19 expression was measured in 70 pairs of ovarian cancer tissue samplescompared with normal controls by real-time quantitative RT-PCR. The effects of H19 on ovarian cancer cells were studied by RNA interference approach. Apoptosis and cell cycle were analyzed by flow cytometry. Cells viability was evaluated using cell counting Kit-8. Our results demonstrated that that H19 silencing inhibited OV90 and SKOV3 OC cell proliferation in vitro. Further investigation into the mechanisms responsible for the growth inhibitory effects by H19 silencing revealed that its knockdown resulted in the induction of cell cycle arrest and apoptosis through certain cell cycle-related and apoptosis-related proteins. Together, our data suggest that LncRNAH19 plays an important role in OC cell proliferation and contributes to a better understanding of the importance of dysregulated lncRNAs in OC progression.     

马兜铃酸通过ERK1/2信号传导途径诱导人脐静脉血管内皮细胞凋亡

 目的 探讨马兜铃酸(AA)是否通过ERK1/2信号传导途径诱导人脐静脉血管内皮细胞(HUVECs)凋亡。方法 通过MTT法检测细胞增殖能力;通过Hoechst 33258荧光染色观察细胞凋亡的形态学改变;通过Annexin V-FITC/PI染色采用流式细胞仪检测细胞凋亡率;通过Western blotting法测定细胞内p-ERK1/2的水平。结果 马兜铃酸呈浓度和时间依赖方式抑制了内皮细胞增殖。马兜铃酸可引起内皮细胞出现凋亡形态学改变,且呈浓度依赖方式增加了内皮细胞凋亡率。同时,马兜铃酸可降低内皮细胞内p-ERK1/2的水平。应用ERK1/2抑制剂PD98095预处理后,细胞内p-ERK1/2的水平与AA (10 mg/L)组相比显著增加,马兜铃酸诱导的内皮细胞凋亡率亦被明显抑制。结论 马兜铃酸可诱导血管内皮细胞凋亡,其机制可能是通过抑制ERK1/2信号传导途径。     

c-Myc-activated long non-coding RNA H19 downregulates miR-107 and promotes cell cycle progression of non-small cell lung cancer

To verify c-Myc can regulate the expression of lncRNA H19 directly in non-small cell lung cancer (NSCLC) and clarify the molecular mechanism on how lncRNA H19 promote the cell cycle progression of NSCLC. The mRNA levels of lncRNA H19 in NSCLC tissues and cells, the adjacent tissues and normal cells were determined by RT-PCR. The expression change of lncRNA H19 in NSCLC cells after transfection with pcDNA3.1-c-Myc or c-Myc-siRNA was determined by RT-PCR, respectively. Targeted role of c-Myc on the promoter of H19 was studied by luciferase reporter assay. Chromosome immune coprecipitation (ChIP) was used to confirm the relationship between c-Myc and H19. MiRNAs that have base-pairing with H19 was predicted by online software. The relationship between H19 and miR-107 was determined by disturbing and overexpressing the expression of H19. The influence of the changes of H19 and miR-107 on cell cycle progression was determined by flow cytometry. The mRNA levels of lncRNA H19 in NSCLC tissues and cells were significantly higher than the adjacent tissues and normal cells, respectively. The expression of H19 increased or decreased accordingly with the overexpression and knockdown of c-Myc. The activity of the promoter of H19 was strengthened by c-Myc. While the expression of miR-107 increased or decreased with the overexpression and knockdown of H19, respectively. The number of cells in G2/M stage decreased significantly with the knockdown of H19 and miR-107 compared with the control group. Our study demonstrates that lncRNA H19, which is induced by c-Myc, is up-regulated in NSCLC. H19 influences the mitotic progression of NSCLC cell lines.     

miR-455-3p靶向HIPK2调控高糖环境下成骨细胞的增殖和凋亡

文题释义： 糖尿病性骨病：糖尿病可导致骨代谢发生改变,表现为骨形成缺陷、成骨细胞数量减少、骨基质形成不足。高糖环境下以成骨细胞功能改变最为显著,高糖诱导成骨细胞是研究糖尿病性骨病的常用细胞模型。 成骨作用：指成骨细胞移至将要合成骨组织的部位,分泌、合成骨胶原及骨蛋白纤维,将钙、磷吸收到纤维孔隙中进行沉淀结晶,形成骨组织的过程。 背景：以往研究表明多种miRNA在骨形成中发挥作用,miR-335-5p可保护成骨细胞免受氧化应激,对枸橼酸铁铵诱导的成骨细胞具有保护作用,但miR-335-5p对高糖环境下成骨细胞增殖和凋亡的影响尚未可知。 目的：探讨miR-455-3p靶向HIPK2对高糖环境下成骨细胞增殖和凋亡的影响。 方法：双荧光素酶报告基因分析法验证miR-455-3p对HIPK2的靶向作用。体外用高糖诱导MC3T3-E1细胞,分为空白组、高糖组、高糖+miR-control组、高糖+miR-455-3p组、高糖+si-control组、高糖+si-HIPK2组、高糖+miR-455-3p+pcDNA组和高糖+miR-455-3p+pcDNA-HIPK2组。qRT-PCR检测miR-455-3p和HIPK2 mRNA的表达,MTT法检测细胞存活率,流式细胞检测细胞凋亡,Western blot检测HIPK2、p-STAT3和STAT3蛋白的表达。 结果与结论：①HIPK2是miR-455-3p的靶基因,miR-455-3p可负性调控HIPK2的表达;②高糖处理可抑制miR-455-3p的表达,促进HIPK2的表达;③过表达miR-455-3p或抑制HIPK2表达均可促进高糖条件下MC3T3-E1细胞的存活和抑制凋亡;④过表达HIPK2可部分逆转miR-455-3p对高糖条件下成骨细胞的存活促进和凋亡抑制作用;⑤miR-455-3p通过调控HIPK2抑制成骨细胞中p-STAT3的表达;⑥结果表明,miR-455-3p通过靶向下调HIPK2抑制高糖诱导的成骨细胞凋亡,促进成骨细胞增殖,这可能与抑制STAT3信号通路有关。 ORCID: 0000-0001-7129-8455(匡嘉兵) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Expression of long non-coding RNA SFTA1P and its function in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is a major type of lung cancer with high morbidity and mortality. Long non-coding RNAs (lncRNAs) have been reported to be important in development and progression of NSCLC. However, the role of lncRNA SFTA1P remains unclear. This study aims to explore the clinical roles, biological function, and mechanism of SFTA1P in NSCLC. SFTA1P expression was estimated by the quantitative real-time polymerase chain reaction (qRT-PCR) of 90 pairs of tissue samples, the Cancer Genome Atlas (TCGA) database and microarray. After overexpressing SFTA1P, NSCLC cell proliferation, cycle, and apoptosis were detected. We found that the expression of SFTA1P was significantly downregulated in NSCLC tissues with high diagnostic value (AUC = 0.87), which was consistent with the results of TCGA and microarray data. For the analysis of clinical features, the results revealed that SFTA1P expression was closely related to the pathological type (P < 0.01). Furthermore, the cell function results suggested that the overexpression of SFTA1P triggered cell cycle arrest in the S-phase (P < 0.05). From a mechanistic perspective, the results showed that the PI3K-AKT signaling pathway was inhibited after overexpression of SFTA1P in NSCLC. Taken together, this work supported that SFTA1P may play a suppressing role in the tumorigenesis of NSCLC by modulating PI3K-AKT signaling pathway to influence cell cycle, which provides a potential and prospective biomarker for NSCLC.     

骨碎补总黄酮联合纳米骨材料促进MC3T3-E1细胞的增殖分化

文题释义： 骨碎补总黄酮：是由水龙骨科植物槲蕨的干燥根茎中提取的有效成分,骨碎补总黄酮能够促进成骨细胞增殖,抑制破骨细胞成熟分化。 Wnt/β-catenin信号通路：是一类高度保守的信号通路,广泛存在于多细胞真核生物中,是皮肤发育过程中出现最早的分子信号,调控毛囊的生长发育和毛囊干细胞的迁移分化。β-catenin作为细胞内信号传导蛋白,是Wnt信号通路激活的一种重要的上皮细胞表面黏附分子,能够进入细胞核内传递Wnt信号,进一步激活靶基因开始转录,启动细胞增殖周期。 背景：前期研究发现,骨碎补总黄酮可促进纳米骨材料表面MC3T3-E1细胞的成骨分化,其作用机制有待进一步研究。 目的：探究骨碎补总黄酮联合纳米骨材料对MC3T3-E1细胞发挥作用的机制。 方法：将MC3T3-E1细胞与纳米骨材料共培养,选取100 mg/L和250 mg/L骨碎补总黄酮进行药物干预,以10 μg/L转化生长因子β刺激为阳性对照组。分组如下：①正常组;②DKK1组：Wnt通路抑制剂DKK1      (0.1 mg/L)阻断Wnt/β-catenin信号通路;③DKK1+转化生长因子β组;④DKK1+100 mg/L骨碎补总黄酮组;⑤DKK1+250 mg/L骨碎补总黄酮组;⑥DKK1+纳米骨+转化生长因子β组;⑦DKK1+纳米骨+100 mg/L骨碎补总黄酮组;⑧DKK1+纳米骨+250 mg/L骨碎补总黄酮组。在干预24,48 h后收获细胞,免疫荧光双染法观察Wnt/β-catenin通路中Wnt与LRP结合情况,Real-time PCR和Western blot检测β-catenin、LRP5、Gsk-3β、Cyclin D1、RUNX2的表达。 结果与结论：①激光共聚焦扫描显微镜下显示DKK1+转化生长因子β组、DKK1+250 mg/L骨碎补总黄酮组、DKK1+纳米骨+转化生长因子β组、DKK1+纳米骨+250 mg/L骨碎补总黄酮组棕黄色染色较明显,表明Wnt与LRP结合较其他组更好;②Real-time PCR和Western blot结果显示,骨碎补总黄酮可促进β-catenin、LRP5、RUNX2的表达,下调GSK-3β的表达,说明骨碎补总黄酮通过激活经典Wnt/β-catenin 信号通路促进成骨细胞增殖分化,且骨碎补总黄酮诱导的基因活化呈剂量依赖性。 ORCID: 0000-0002-2031-8644(李晋玉) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

Long non-coding RNA ncRuPAR regulates gastric cancer cell proliferation and apoptosis via phosphoinositide 3-kinase/protein kinase B signaling

Objective: To determine the effect and mechanism of the long non-coding RNA (lncRNA) ncRuPAR (non-protein coding RNA, upstream of coagulation factor II thrombin receptor [F2R]/protease-activated receptor-1 [PAR-1]) in human gastric cancer.Methods: HGC-27-ncRuPAR overexpression and MGC-803-ncRuPAR-RNAi knockdown gastric cancer cell lines were established. We assessed the effect of ncRuPAR on cell proliferation, apoptosis, migration, and invasion using Cell Counting Kit 8, flow cytometry, scratch and transwell assays, respectively. Differentially expressed genes in HGC-27-ncRuPAR overexpression and HGC-27-empty vector cell lines were identified using Affymetrix GeneChip microarray analysis. Ingenuity Pathway Analysis (IPA) of the microarray results was subsequently conducted to identify ncRuPAR-enriched pathways, followed by validation using real time-quantitative PCR (RT-qPCR). As one of the top enriched pathways, phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway was further examined by western blotting to determine its role in ncRuPAR-mediated regulation of gastric cancer pathogenesis.Results: ncRuPAR inhibited human gastric cancer cell proliferation and induced G1/S phase arrest and apoptosis, but did not affect migration or invasion in vitro. Overexpression of ncRuPAR in vitro was found to inhibit its known target PAR-1, as well as PI3K/Akt signaling. The downstream targets of PI3K/Akt, cyclin D1 was downregulated, but there was no change in expression level of B-cell lymphoma 2 (Bcl-2).Conclusions: We showed that lncRNA-ncRuPAR could inhibit tumor cell proliferation and promote apoptosis of human gastric cancer cells, potentially by inhibiting PAR-1, PI3K/Akt signaling, and cyclin D1. The results suggest a potential role for lncRNAs as key regulatory hubs in GC progression.     

成纤维细胞生长因子受体4基因沉默对口腔鳞癌细胞增殖和凋亡的影响及其作用机制

目的：探讨成纤维细胞生长因子受体4（FGFR4）基因沉默介导RAS/RAF/MAPK信号通路对口腔鳞癌细 胞增殖和凋亡的影响。方法：收集本院2018 年4 月至2019 年8 月收治的口腔鳞癌患者32 对肿瘤组织样本和癌旁 非癌组织,利用免疫组织化学和免疫印迹检测FGFR4蛋白的表达;免疫印迹检测正常口腔上皮细胞HOEC和口 腔鳞癌细胞（HSC-2、CAL-27 和SACC-83）中FGFR4蛋白的表达水平。通过基因沉默技术下调FGFR4表达,利 用qRT-RCR 和免疫印迹检测SACC-83 细胞的FGFR4表达的变化,克隆形成实验检测SACC-83 细胞的增殖能力, 流式细胞术检测SACC-83 细胞的凋亡和细胞周期,免疫印迹检测SACC-83 细胞的RAS/RAF/MAPK信号通路。 结果：与癌旁非癌组织相比,口腔鳞癌组织的FGFR4表达升高;与正常口腔上皮细胞HOEC相比,口腔鳞癌细 胞（HSC-2、CAL-27 和SACC-83）的FGFR4表达量高;下调FGFR4表达可抑制SACC-83 细胞的增殖,抑制抗 凋亡蛋白Bcl-2 的表达,上调凋亡蛋白Bax 的表达,促进SACC-83 细胞的凋亡,同时抑制周期蛋白P21 的表达, 使SACC-83 细胞停滞于G0/G1 期;下调FGFR4表达可抑制RAS、RAF及p-MEK和p-ERK1 蛋白的表达。结论： 下调FGFR4表达可抑制口腔鳞癌细胞增殖,促进细胞凋亡,使其停滞于G0/G1 期,可能与抑制RAS/RAF/MAPK 信号通路有关。