Deregulation of FGFR1 and CDK6 oncogenic pathways in acute lymphoblastic leukaemia harbouring epigenetic modifications of the MIR9 family

The role of epigenetic mechanisms in the regulation of microRNAs (miRNAs) with a tumour‐suppressor function in human neoplasms has recently been established. Several miRNAs have been found to be inappropriately regulated by DNA methylation in patients with acute lymphoblastic leukaemia (ALL). We analysed the methylation status of the three members of the MIR9 family (MIR9‐1, MIR9‐2 and MIR9‐3) in a uniformly treated cohort of 200 newly diagnosed ALLs. MIR9 was methylated in 54% of the patients and was associated with downregulation of MIR9 (P < 0·01). Hypermethylation of MIR9 was an independent prognostic factor for disease‐free survival, overall survival and event‐free survival in a multivariate analysis (P < 0·01). Epigenetic downregulation of MIR9 induced upregulation of its targets, FGFR1 and CDK6, while treatment of ALL cells with FGFR1 (PD‐173074) and CDK6 (PD‐0332991) inhibitors induced a decrease in cell proliferation and an increase in apoptosis of ALL cells. Our results indicate that the MIR9 family is involved in the pathogenesis and clinical behaviour of ALL and provide the basis for new therapeutic strategies in the treatment of ALL, targeting the epigenetic regulation of miRNAs and/or the FGFR1 or CDK6‐RB pathway directly.     

miR-132通过靶向KLF7抑制鼻咽癌细胞增殖和迁移

目的探讨miR-132靶向负调节Kruppel样因子7(KLF7)表达对鼻咽癌(NPC)细胞增殖和迁移的影响。荧光定量PCR(qPCR)检测NPC组织和细胞及正常鼻咽组织和细胞中miR-132表达,Western blot检测KLF7蛋白表CNE-2Z细胞增殖、迁移和侵袭能力,裸鼠成瘤实验、免疫组化法检测CNE-2Z细胞体内生长、瘤内血管情况。结果鼻咽癌组织较正常鼻咽组织miR-132表达水平降低,KLF7蛋白表达增加;鼻咽癌CNE-2Z细胞较正常鼻咽上皮细胞NP69 miR-132表达水平降低,KLF7蛋白表达增加(P<0.05)。TargetScan预测及双荧光素酶检测显示,KLF7是miR-力,迁移和侵袭数量,肿瘤体积及微血管密度(MVD)显著降低(P<0.05),KLF7-OE组结果均相反(P<0.05)。相比miR-132 mimics组,miR-132 mimics+KLF7-OE组KLF7蛋白表达水平,36、48、60 h细胞增殖活力,迁移和侵袭数量、肿瘤体积及MVD显著升高(P<0.05)。结论 miR-132可通过负调控KLF7表达,影响NPC...     

脓毒症患者血小板微粒与微RNA-155水平的变化规律及意义

目的 探讨血小板微粒(PMPs)和微RNA(miRNA)-155与脓毒症患者病情和预后的关系.方法 选择脓毒症患者61例,根据28 d预后将其分为存活组和死亡组;根据病情严重程度将其分为脓毒症组、严重脓毒症组及脓毒症休克组.比较各组患者APACHEⅡ评分,使用流式细胞仪进行PMPs定量,利用实时荧光定量PCR检测患者血浆miRNA-155水平.Spearman相关性分析确认PMPs水平、miRNA-155水平与患者APACHEⅡ评分的关系.结果 与存活组比较,死亡组患者PMPs水平较高(P＜0.05).脓毒症、严重脓毒症、脓毒症休克3组患者的PMPs水平比较,差异均有统计学意义(P＜0.05).死亡组患者的miRNA-155水平相对于存活组提高(P＜0.05),严重脓毒症组及脓毒症休克组患者的miRNA水平较脓毒症组明显提高(P＜0.05).患者PMPs水平及miRNA-155水平均与APACHEⅡ评分呈正相关,同时患者PMPs水平与microRNA-155水平也呈正相关.PMPs水平联合miRNA-155水平预测脓毒症患者死亡的ROC曲线下面积达0.831.结论 脓毒症患者的PMPs水平及miRNA-155水平与脓毒症患者的病情及预后有关,miRNA-155可能引起脓毒症患者的PMPs水平增高.     

MicroRNA-148b and microRNA-152 reactivate tumor suppressor genes through suppression of DNA methyltransferase-1 gene in pancreatic cancer cell lines

Overexpression of DNA methyltransferase 1 (DNMT-1) is observed mostly in pancreatic cancer and it can cause tumor suppressor genes silencing in this disease. Recent studies suggest that abnormal expressions of microRNAs (miRs) are involved in pathogenesis of different types of human cancers including pancreatic cancer. In this study we aimed to investigate the effect of miR-148b and -152 on reverting the tumorigenic phenotype of pancreatic cancer cell lines. In order to investigate whether miR-148b and -152 are involved in the regulation of DNMT-1, luciferase reporter assay was used and confirmed that the DNMT-1 mRNA could be a target for miR-148b and miR-152. Furthermore, overexpression of miR-148b and -152 in pancreatic cancer cell lines (MIA PaCa-2 and AsPC-1) decreased DNMT-1 expression (53% and 59% respectively), returned DNA methylation to normal patterns and induced re-expression of tumor suppressor genes, like BNIP3 (4.7- and 3.8-fold) and SPARC (5.3- and 2.9-fold) for miR-148b and -152 respectively. Moreover, the introduced miR-148b and -152 could inhibit the proliferation of MIA PaCa-2 (35% and 37% respectively) and AsPC-1 (39% and 40% respectively) cell lines. The apoptosis rates of MIA PaCa-1 after treatment with miR-148b and -152 were 10% and 8% respectively; while these rates in AsPC-1 were 16% and 11% respectively. Conclusively these findings mean that miRs that are targeting DNMT-1 and modifying methylation status of tumor suppressor genes such as BNIP3 and SPARC can be applied in killing the pancreatic cancer cells and decreasing the tumorigenicity of these cells.     

MicroRNAs：非小细胞肺癌分子诊断和靶向治疗重要的靶标

microRNAs（miRNAs）是一类长度约为18~22个核苷酸、在动植物中广泛存在、高度保守的小分子非编码RNA。非小细胞肺癌（NSCLC）与其起源的相应正常细胞之间存在miRNAs表达谱的差异,不同类型或不同进展状态非小细胞肺癌之间miRNAs表达谱也有其特异性,为肺癌的早期诊断和鉴别诊断提供了候选指标。利用miRNAs调控靶基因表达的原理,还可以发现或设计更优化的肿瘤治疗模式。深入研究miRNAs在非小细胞肺癌中的作用和价值,将为筛选NSCLC分子诊断指标并据此建立靶向干预策略奠定基础。     

生信分析筛选间充质干细胞外泌体中与血管新生相关微小RNAs及黄芪甲苷干预的研究

目的 探讨黄芪甲苷（AS-Ⅳ）对生信分析筛选出的人脐带血间充质干细胞（hUCBMSCs）外泌体（Exos）中与血管生成相关微小RNAs（miRNAs）表达的影响。方法 通过生信分析筛选hUCBMSCs-Exos中与血管生成高度相关的miRNAs。将hUCBMSCs随机分为实验组和对照组。实验组给予含300 mg·L^-1 AS-Ⅳ的DMEM/F12培养基培养24 h,对照组加入等量磷酸盐缓冲溶液培养24 h。用二辛丁酸法检测Exos浓度,用实时荧光聚合酶链反应法检测miRNAs的表达水平。结果 实验组和对照组的外泌体浓度分别为（1.22±0.02）和（0.88±0.03）μg·μL^-1,差异有统计学意义（P<0.01）。miRNA-126-3p、miRNA-21、miRNA-214-5p、miRNA-126-5p、miRNA-145、miRNA-16-5p和miRNA-195-5p在实验组的表达量分别为6.69±0.57,1.06±0.16,1.06±0.21,0.68±0.05,0.51±0.17,0.48±0.14和0.46±0.10...     

miR-125a在支气管哮喘患儿血浆中的表达及其临床意义

目的 评估miR-125a在儿童支气管哮喘中的表达及其与疾病患病风险、急性发作风险、严重程度与炎症水平的关系。 方法 连续纳入急性发作期、缓解期支气管哮喘患儿和匹配的健康儿童各70例,收集受试者实验室检测及肺通气功能检测结果。采用实时荧光定量聚合酶链式反应检测受试者血浆miR-125a表达水平,采用酶联免疫吸附试验检测受试者血浆中肿瘤坏死因子(TNF-α),白细胞介素(IL-)1β,IL-6和IL-17的水平。 结果 miR-125a在急性发作期支气管哮喘患儿血浆中表达水平显著低于缓解期支气管哮喘患儿(P<0.001)和健康儿童(P<0.001),在缓解期支气管哮喘患儿血浆中表达水平也显著低于健康儿童(P=0.022)。受试者工作特征曲线表明,miR-125a能显著区分3组受试者。同时,miR-125a表达水平与急性发作期支气管哮喘患儿疾病严重程度(P<0.001)、TNF-α(P=0.005)、IL-1β(P=0.007)及IL-17(P=0.037)呈显著负相关,与第1秒用力呼气容积(FEV₁)与用力肺活量(FVC)之比(FEV₁/FVC)(P<0.001)和FEV₁实测/预测值百分比(P<0.001)呈显著正相关。miR-125a表达水平还与缓解期支气管哮喘患儿TNF-α(P=0.032)及IL-17(P=0.001)表达水平和健康儿童IL-6(P=0.024)及IL-17(P=0.026)表达水平呈显著负相关。 结论 循环miR-125a表达水平与儿童支气管哮喘患病风险、急性发作风险、严重程度及炎症水平呈显著负相关。     

急性脑梗死早期血清lncRNA XIST和miR-92a水平与神经功能缺损程度的关系

目的:探讨急性脑梗死患者早期外周血清中长链非编码RNAX染色体失活特异转录物(lncRNA XIST)和微小核糖核酸(miRNAs)-92a水平与神经功能缺损程度的关系及意义。方法:急性脑梗死患者70例纳入梗死组,健康体检者48例纳入对照组。根据美国国立卫生研究院卒中量表(NIHSS)对患者进行神经功能缺损程度评分,NIHSS<7分者纳入轻型梗死组,NIHSS≥7分者纳入重型梗死组。于脑梗死后48 h和(或)14 d,检测脑梗死患者血清lncRNA XIST和miR-92a水平,并进行不同组别间的比较。Pearson相关及Logistic回归分析lncRNA XIST和miR-92a水平与梗死患者NIHSS评分间的关系。结果:脑梗死后48 h时,梗死组患者血清中lncRNA XIST相对表达量低于对照组,miR-92a的水平高于对照组(均P<0.05)。脑梗死后48 h时,重型梗死组患者血清中lncRNA XIST相对表达量低于轻型梗死组,miR-92a的水平为高于轻型梗死组(均P<0.01)。发病后14 d患者血清中lncRNA XIST相对表达量高于发病后48 h,miR-92a的水平低于发病后48 h(均P<0.01)。急性脑梗死患者血清中lncRNA XIST水平与NIHSS评分负相关,miR-92a水平与NIHSS评分正相关(均P<0.01)。lncRNA XIST和miR-92a都是急性脑梗死早期神经功能缺损程度的影响因素(均P<0.01)。结论:脑梗死早期血清lncRNA XIST和miR-92a与急性脑梗死早期神经功能缺损程度密切相关,可望作为脑梗死后潜在的新分子干预靶点。     

结肠癌相关miRNA的转化机制

结肠癌的发生是多阶段、多步骤的病变过程,是从良性息肉-浸润性腺癌-癌远处转移的演变过程.这些病理变化与编码蛋白质的原癌基因和肿瘤抑制基因的异常激活或失活有关.但最近关于微小RNA(microRNAs,miRNAs)的研究改变了我们对非编码蛋白质基因在肿瘤发生中的作用的理解.在结肠癌组织中多种miRNAs的表达水平不同,...