Expression of the monoclonal antibody HECA-452 defined E-selectin ligands in Langerhans cell histiocytosis

The cutaneous lymphocyte associated antigen (CLA) recognized by the monoclonal antibody (moAb) HECA-452 plays a major role in the homing of lymphocyte subpopulations to the skin by binding to E-selectin on dermal microvessels. The factors responsible for the immigration of Langerhans cells (LC) and their precursors into the skin are still unknown, but because normal resting LC are also capable of expressing CLA, the antigen was proposed as a candidate LC-homing structure. To gain insight into these mechanisms, the expression of HECA-452 on neoplastic LC within and outside the skin was investigated in paraffin-embedded sections from 44 patients with localized and disseminated forms of Langerhans cell histiocytosis (LCH) presenting with proliferating cells positive for CD45, CD1a, S100 and HLADR. Irrespective of the clinical presentation or the type of organ involved, HECA-452-positive LC were detected in all biopsies tested (range 5->90%). The most prominent HECA-452 reactivity was observed in skin lesions and in areas with accumulations of eosinophilic granulocytes. Our data provide evidence for a heterogeneous expression of sLe^x/sLe^a structures in various stages of activated and/or differentiated LCH cells. Remarkably, CLA-antigen expression on LCH-cells was not restricted to cutaneous sites. In view of recent findings on the expression of HECA-452 on resting epidermal LC, our data are compatible with the view that local cytokine production by keratinocytes or cells from the surrounding infiltrate induce and/or modulate CLA expression on LC in both cutaneous and extra-cutaneous sites.This work is dedicated to Professor Dr. Thaddäus Radaszkiewicz, who died in September 1995     

miR-31调控PAX9对肺癌细胞增殖和侵袭能力的影响

目的 研究miR-31调控配对盒基因9(PAX9)对肺癌细胞侵袭和增殖能力的影响.方法 运用免疫组织化学检测肺癌组织和癌旁正常组织PAX9蛋白的表达;运用荧光定量PCR检测肺癌组织、癌旁正常组织和肺癌细胞株M14中miR-31的表达.通过转染miR-31-inhibitor下调M14细胞中miR-31的表达,双荧光素酶活性检测miR-31对PAX9转录活性的影响;Transwell侵袭实验检测miR-31对M14细胞的侵袭能力的影响;平板克隆实验检测miR-31对M14细胞的增殖能力的影响.结果 与癌旁正常组织比较,PAX9蛋白在肺癌组织中表达降低,miR-31在肺癌组织中表达明显升高(P<0.05);双荧光素酶活性检测结果显示,miR-31可以直接调控PAX9的转录活性;抑制miR-31的表达后,肺癌细胞株M14的PAX9蛋白表达水平上调,侵袭和转移能力明显降低(P<0.05).结论 miR-31靶向PAX9的表达,从而调控肺癌细胞的侵袭和增殖能力.     

骨肉瘤患者血浆中microRNA-181b水平的检测

目的检测骨肉瘤患者血浆中microRNA-181b (miR-181b)的水平变化,初步探讨其临床意义.方法选择25例骨肉瘤患者和30例健康对照,采用茎-环定量逆转录聚合酶链反应(RT-PCR)方法检测血浆中miR-181b的水平,并分析miR-181b与骨肉瘤临床病理指标之间的关系.结果骨肉瘤患者血浆中miR-181b水平(0.62±0.25)与正常对照组(0.41±0.18)相比明显升高,差异具有统计学意义(P＜0.01);骨肉瘤患者血浆中miR-181b的水平与患者性别、年龄、肿瘤大小、肿瘤部位、病理类型及Enneking分期均无明显相关性(P＞0.05).结论骨肉瘤患者血浆中miR-181b水平增高,miR-181b可能参与了骨肉瘤的发病过程.     

MicroRNA-21与胃癌关系的研究进展

胃癌的发生和发展是一个复杂、多步骤的过程,涉及原癌基因和抑癌基因的遗传失调,最近研究较多的微RNA(miRNA)起着重要作用。miRNA是一种长为21²3个核苷酸的非编码RNA分子,参与细胞增殖、凋亡和分化以及恶性肿瘤的发生,是重要的调节器。近些年来,人们针对miR-21在恶性肿瘤中的表达研究最为广泛深入,该文就miR-21与胃癌的关系进展予以综述。     

人类微小RNA-208a的靶基因预测及生物学过程和信号通路的生物信息学分析

目的 采用生物信息学技术预测人类微小RNA-208a(hsa-miRNA-208a)靶基因及分析其可能参与的生物学过程及信号通路.方法 应用miRbase数据库和UCSC Genome Browser分析工具获取hsa-miRNA-208a的染色体定位、碱基序列和物种保守性等基本信息,利用miRanda、miRDB、TargetScan和miRwalk进行靶基因预测,采用miRwalk网站对靶基因取交集,合并有文献支持和经实验证实的靶基因作为基因集,对基因集进行功能富集分析(GO分析)和信号通路分析.结果 hsa-miRNA-208a序列在各物种间高度保守.对TargetScan、miRDB、miRanda和miRwalk预测的靶基因进行交集后共得到16个靶基因(CPEB2、CSNK2A2、DLD、MTF2、FBXO28、LEP、SMAD4、NLK、GPR88、VPS13D、ZNF215、CHD9、SLC7A5、C19orf44、SLC45A3、MAP4K4),miRwalk、DIANA Lab TarBase检索到已验证的靶基因分别为12个(CASP3、MRGPRX3、FPRL1、MED13、IL10、GATA4、HSH2D、MYH6、MYH7、TNNI3、CDKN1A、PAK3)和3个(SOX6、MTM1、TAB3).GO分析结果显示靶基因富集于心室肌组织发育、心室形态发育、心腔发育、心肌发育、心脏发育以及细胞发育等生物学过程(P＜0.05);信号通路分析结果显示靶基因富集于黏附连接、紧密连接、心肌收缩、Wnt信号通路以及病毒性心肌炎、肥厚型心肌病和扩张型心肌病的相关信号通路中(P＜0.05).结论 hsa-miRNA-208a参与心脏生长发育的生物学过程,与心肌疾病的发生密切相关.     

miR-429 expression in bladder cancer and its correlation with tumor behavior and clinical outcome

We previously showed that microRNA-429 (miR-429) played an important role in epithelial–mesenchymal transition (EMT) of urothelial cell carcinoma of the bladder. We herein evaluated the expression of miR-429 in bladder cancer and its potential relevance to clinicopathological characteristics and patient survival. Relative expression levels of miR-429 in surgical bladder cancer tissue specimens obtained from 76 patients with bladder cancer were measured by chromogenic in situ hybridization. miR-429 expression was significantly higher in specimens from alive patients than expired patients in both of 5-year overall survival (OS) (0.59 ± 0.09 vs. 0.27 ± 0.12; p < 0.05) and 5-year recurrence-free survival (RFS) (0.63 ± 0.10 vs. 0.33 ± 0.10; p < 0.05). The univariate Cox proportional hazards analysis revealed that tumor grade, stage, and miR-429 expression were significantly associated with patient survival. In multivariate analysis, tumor stage and miR-429 expression were significantly associated with 5-year OS (hazard ratio [HR] 4.70, p < 0.001) and 5-year-RFS (HR 2.20, p < 0.05). The Kaplan–Meier analysis showed that patients with miR-429 expression had significantly better 5-year OS and 5-year RFS rates than those without miR-429 expression (84.4% vs. 61.4%, p < 0.05 and 71.9% vs. 45.5%, p < 0.05, respectively). miR-429 may be considered as an adjunctive prognostic marker in addition to tumor grade and stage in bladder cancer.     

MicroRNA4293基因多态性与女性肝癌易感性相关

目的研究microRNA-4293的SNP(rs12220909)与原发性肝癌易感性的关系。方法收集人体血液样本1788份,样本取自健康对照者884名,原发性肝癌患者904例。抽提血液样本中基因组DNA,应用MassARRAY法对microRNA-4293的SNP(rs12220909)位点进行基因分型;应用二元逻辑回归法分析microRNA-4293的SNP(rs12220909)位点与肝癌易感性之间的关系。结果女性肝癌患者microRNA-4293的SNP(rs12220909)中CC基因型较健康对照组显著增加(P=0.04),在HBV相关肝癌中显著性进一步增加(P=0.02),提示CC基因可能增加女性肝癌尤其是HBV相关肝癌的易感性。而在男性患者中,microRNA-4293的SNP(rs12220909)与肝癌易感性并不相关(P=0.33)。结论microRNA-4293的SNP(rs12220909)与女性肝癌易感性相关。     

miR-21反义寡核苷酸增强地西他滨体外抗白血病效应

目的:研究miR-21反义寡核苷酸(anti-miR-21 oligonucleotide,AMO)对地西他滨(decitabine,DCA)抗白血病效应的影响及可能机制。方法:将AMO和无义寡核苷酸(scramble oligonucleotide,SCR)通过脂质体转染导入HL-60细胞,实时荧光定量PCR(real-time PCR)验证转染效率,再分别与DCA 0.5、2.0和4.0μmol/L作用48 h。Real-time PCR分别检测人周期节律蛋白3(h Per3)mRNA表达,Annexin V/PI法检测凋亡,流式细胞术检测CD117和CD11b平均荧光强度(MFI)。结果:AMO转染组miR-21表达(0.35±0.07)低于空白组(0.71±0.07)和SCR转染组(0.66±0.05),差异有统计学意义(P0.05)。AMO转染组的HL-60细胞DCA的IC50低于空白组和SCR转染组(P0.01)。同一浓度下,AMO组的早期凋亡率、CD11b的MFI和h Per3 mRNA均高于同一浓度药物作用的空白组和SCR组,CD117 MFI均低于同一浓度药物作用的空白组和SCR组,差异均具有统计学意义(P0.01)。结论:AMO能显著促进DCA体外抗白血病效应,其机制可能与其协助激活h Per3的表达有关。     

Serum and urinary lipoproteins in the human nephrotic syndrome: evidence for renal catabolism of lipoproteins

The urinary excretion of lipoproteins and the possibility of catabolic alterations on glomerular filtration were investigated in four nephrotic subjects differing in etiology, serum lipoprotein profile, and 24 hr urinary output of protein and lipids. The apolipoproteins and lipoproteins of urine were compared with those of serum with respect to distribution profile, physical properties, and composition. Lipoprotein particles resembling the serum very low, intermediate, low, and high density lipoproteins (VLDL, IDL, LDL, and HDL, respectively) in density, particle size, and morphology were isolated from the urine. As expected from molecular sieving effects during glomerular filtration, the urinary HDL were more abundant than the lower density lipoproteins even when the plasma LDL was elevated markedly. However, little sieving effect was seen within the urinary HDL, which comprised a broad spectrum of particle sizes including the larger HDL₂, whose average diameter was similar to that of the plasma HDL. A sieving effect was not seen in the urinary LDL, except for a greatly increased proportion, about 20% of total particles, of HDL-like species. Intact apolipoproteins were not found in the concentrated urinary fraction isolated by ultrafiltration between the limits of 10⁴ and 5 × 10⁴ daltons. On the basis of immunoreactivity, gel electrophoresis, and amino acid composition, apolipoproteins B and AI are the major and minor proteins, respectively, of urinary LDL, and apo B is the major protein of the urinary IDL and VLDL. Apolipoproteins AI, AII, CI, CIII, and possibly AIV were isolated from the urinary HDL. As much as 20% of the protein moiety of the urinary HDL appeared to be large apolipoprotein fragments with molecular weights and isoelectric points similar to those of apo CII and apo CIII. The fragments were derived in part from apo AI, the least acidic form of which was lost preferentially. The lower density classes of urinary lipoproteins also appeared to have lost apo E and apo C's and to have undergone partial proteolysis. Apparently, the surface-exposed, readily exchangeable apolipoproteins are subject to proteolytic degradation upon glomerular filtration.     

miR-1-3p在食蟹猴心肌损伤中表达水平及其与CK-MB、cTnI和NT-proBNP的相关性

目的探讨循环miR-1-3p在多柔比星诱导食蟹猴心肌损伤模型中的表达水平和诊断价值。方法选取3~5岁普通级雄性食蟹猴17只,随机分为对照组(7只)和实验组(10只),静脉注射盐酸多柔比星注射液(2.5 mg/kg)4周,收集外周血浆标本和心脏组织标本,应用液相芯片技术测定肌酸激酶同工酶(CK-MB)、肌钙蛋白I(cTnI)和NT-proBNP表达水平,且应用实时定量PCR技术测定miR-1-3p表达的水平,并进行相关性分析。结果成功建立药物致食蟹猴心肌损伤模型;两组CK-MB、cTnI和NT-proBNP水平比较差异有统计学意义(P<0.05);实验组miR-1-3p的表达水平在给药4周后较给药前和对照组相比明显升高(P<0.05);miR-1-3p的表达水平与CK-MB、cTnI和NT-proBNP改变呈现了相同的升高趋势,且二者之间具有明显相关性。结论miR-1-3p可作为药物引起心肌损伤的生物学诊断标志物之一。