转录因子Kaiso下调人乳腺癌细胞系中lncRNA MEG3的表达

目的 探讨转录因子Kaiso是否调节人母系表达基因3(MEG3)表达及其在乳腺癌的发生发展中发挥重要作用.方法 RT?qPCR检测正常乳腺细胞系MCF?10A和多个乳腺癌细胞系MCF?7、BT?474、MDA?MB?435、MDA?MB?231中Kaiso和MEG3的表达;蛋白印迹检测Kaiso在各种细胞中的表达;免疫...     

乳腺癌相关长链非编码RNA的研究进展

乳腺癌在女性恶性肿瘤的发病率中高居第一,长链非编码RNA(long non-coding RNA,lncRNA)和乳腺癌的发生发展、转移和预后相关。lncRNAs可与miRNA、mRNA或蛋白相互作用来调节相关基因的表达能力,从而影响乳腺癌的治疗效果及预后情况。     

肝细胞癌相关长链非编码RNA差异表达的研究

目的分析肝细胞癌与正常肝组织中差异表达的长链非编码RNA(lncRNA),发现可能与肝细胞癌发生发展相关的特异lncRNA。方法采用lncRNA表达谱芯片技术检测5例肝细胞癌和其配对癌旁正常组织中的lncRNA,筛选出差异表达的lncRNA。运用分层聚类分析,对所有的lncRNA进行分类。最后从中选择10个lncRNA用RT-PCR进行定量验证。结果有1359个lncRNA表达水平出现显著性差异,差异倍数〉2倍。其中629个显著上调,占46.2%,其中125个表达5倍以上;730个显著下调,占53.7%,其中110个表达5倍以上。聚类分析(1)基因间lncRNA有11706条,其中与已知编码基因相距300 kb的lncRNA共有352条。(2)增强子型lncRNA共有1802条,其中与已知编码基因相距300 kb的lncRNA共有42条。(3)HOX基因簇共有101条。RT-PCR结果有8条lncRNA表达趋势与基因芯片结果一致。对癌组和癌旁组进行检验,有6条差异有统计学意义(P0.05)。结论与癌旁组织比较,lncRNA肝细胞癌组织中的表达水平明显改变,可能与肝细胞癌的发生发展有关。     

长链非编码RNA在结肠癌组织与癌旁组织中的表达研究

目的:筛选出结肠癌差异表达的长链非编码RNA(lncRNA),并分析其在结肠癌组织和相应癌旁组织中的差异表达情况。方法:从lncRNAtor数据库下载结肠癌组织中差异表达的lncRNA数据(Colon adenocarcinoma:Person neoplasm cancer status),包含36例结肠癌组织及29例正常结肠组织,以P0.01且差异表达倍数大于2或小于0.5的条件筛选出lncRNA,并用real-time PCR进一步验证其在60对结肠癌组织及癌旁组织中的表达情况。结果:分析结肠癌lncRNA数据发现,与正常组织相比,结肠癌组织共有50个lncRNA差异表达,其中28个高表达,22个低表达(P0.01)。筛选的4个lncRNA在60对结肠癌及癌旁组织标本中的验证结果为:HNF1AAS1和ZDHHC8P1的表达均上调(P0.01),SUZ12P表达下调(P0.05)。临床Ⅰ-Ⅱ期结肠癌组织中HNF1AAS1的表达水平明显低于Ⅲ-Ⅳ期(P0.05),ROC曲线分析显示,HNF1A-AS1、ZDHHC8P1和SUZ12P诊断结肠癌的ROC曲线下面积(AUC)分别为0.729(敏感性为78%,特异性为67%)、0.617(敏感性为68%,特异性为55%)和0.689(敏感性为65%,特异性为55%)。结论:长链非编码HNF1A-AS1和ZDHHC8P1在结直肠癌组织中表达上调,SUZ12P在结直肠癌组织中表达下调,其表达水平可能与结肠癌的发生存在关联。     

急性髓系白血病患者血清lncRNA DANCR、lncRNA SNHG7表达及与病理特征、预后的关系

目的 探究急性髓系白血病(AML)患者血清长链非编码RNA抗分化非编码RNA(lncRNA DANCR)、长链非编码RNA小核仁RNA宿主基因7(lncRNA SNHG7)表达及与病理特征、预后的关系。方法 选取2017年10月至2018年10月期间在本院就诊后出院的120例AML患者记为AML组,另选取120例在本院体检的志愿者记为对照组。观察并记录所有参与者年龄、性别、AML形态学分型等临床指标。实时荧光定量PCR(qRT-PCR)方法检测lncRNA DANCR、lncRNA SNHG7表达量;t检验方法分析lncRNA DANCR、lncRNA SNHG7表达差异及其与病理特征的关系;Pearson相关性分析二者表达水平的相关性;根据二者表达水平均值将AML患者分为lncRNA DANCR高表达组(n=65)和lncRNA DANCR低表达组(n=55)及lncRNA SNHG7高表达组(n=74)和lncRNA SNHG7低表达组(n=46);Kaplan-Meier法进行生存分析。结果 AML患者lncRNA DANCR、lncRNA SNHG7水平显著高于对照组(P<...     

lncRNA在心血管疾病中作用的研究进展

正长链非编码RNA(long noncoding RNA,lnc RNA)是一类转录本长度超过200个核苷酸的RNA分子,是RNA聚合酶II转录的副产物,起初它被认为是基因组转录的"噪音",不具有生物学功能。近些年来的研究表明,lnc RNA参与了X染色体沉默,基因组印记以及染色质修饰,转录激活,转录干扰,核内运输等多种重要的调控过程,lnc RNA的这些调控作用     

肺腺癌患者血清中lncRNA SFTA1P的表达及其诊断价值

目的评价肺组织特异的lncRNA SFTA1P在肺腺癌患者血清中的表达情况及其对肺腺癌的诊断价值。方法收集本院132名肺腺癌患者术前血清标本,同时选取年龄、性别匹配的73名健康体检者作为对照。采用实时定量PCR方法检测血清中lncRNA SFTA1P的表达水平,并采用受试者工作曲线分析其对肺腺癌的诊断价值。结果肺腺癌患者血清中lncRNA SFTA1P的水平显著高于健康对照(P<0.001),ROC曲线下面积为0.627。结论lncRNA SFTA1P是潜在的肺腺癌诊断的血清学标志物。     

lncRNA与miRNA相互作用对疾病的影响

长链非编码RNA(lncRNA)与微小RNA(miRNA)之间存在相互调控关系。lncRNA可作为一种竞争性内源性RNA(ceRNA)与miRNA相互作用,参与靶基因的表达调控,反之,miRNA可通过RNA诱导沉默复合物(RISC)调控lncRNA发挥生物学功能,两者共同参与多种疾病的发生。     

lncRNA MALAT1通过上调miR-30a-5p促进乳腺癌细胞增殖

目的 探讨lncRNA MALAT1靶向miR-30a-5p对乳腺癌细胞增殖的影响。方法 CCK-8实验检测lncRNA MALAT1对乳腺癌细胞MCF7增殖的影响。建立MCF7荷瘤小鼠模型检测lncRNA MALAT1对荷瘤小鼠肿瘤生长的影响。通过生物信息学网站预测miR-30a-5p是否与lncRNA MALAT1结合,并通过双荧光素酶报告基因实验进行验证。Real time-qPCR检测lncRNA MALAT1过表达对miR-30a-5p表达的影响。CCK-8实验检测lncRNA MALAT1过表达载体和miR-30a-5pmimics共转染到MCF7细胞对细胞增殖的影响。结果过表达lncRNA MALAT1促进MCF7细胞的体外增殖及MCF7细胞荷瘤小鼠肿瘤生长。lncRNA MALAT1与miR-30a-5p结合,且过表达lncRNA MALAT1下调miR-30a-5p的表达。结论 lncRNA MALAT1通过下调miR-30a-5p表达促进乳腺癌细胞增殖。     

GABRP基因在基底样三阴乳腺癌中的差异表达

目的:探索GABRP基因在基底样三阴乳腺癌(basal-like Triple negative breast cancer,BLTNBC)中的表达,为临床提供预后诊断分子标志物.方法:通过生物信息学分析比较GABRP基因的表达;采用定量PCR方法检测GABRP基因在乳腺癌细胞系及104例乳腺癌石蜡样本中的差异表达,并采用相关性分析、卡方检验及Fisher精确概率法分析GABRP基因与细胞增殖、淋巴结转移、ER及HER-2基因表达的相关性.结果:生物信息学分析和定量PCR结果显示GABRP基因在三阴性乳腺癌(triple negative breast cancer,TNBC)及BLTNBC中显著性高表达(p＜0.05);在non-TNBC,TNBC,BLTNBC中,其表达与细胞增殖、HER-2表达无明显相关性(P＞0.05),而在BLTNBC中与淋巴结转移状态有明显相关性(P＜0.05),在non-TNBC巾与ER表达有明显相关性(P＜0.05).结论:GABRP基因可作为BLTNBC的有潜力的预后诊断分子标志物.     

RNA-seq analysis identified hormone-related genes associated with prognosis of triple negative breast cancer

Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer that currently lacks effective biomarkers and therapeutic targets required to investigate the diagnosis and treatment of TNBC. Here we performed a comprehensive differential analysis of 165 TNBC samples by integrating RNA-seq data of breast tumor tissues and adjacent normal tissues from both our cohort and The Cancer Genome Atlas (TCGA). Pathway enrichment analysis was conducted to evaluate the biological function of TNBC-specific expressed genes. Further multivariate Cox proportional hazard regression was performed to evaluate the effect of these genes on TNBC prognosis. In this report, we identified a total of 148 TNBC-specific expressed genes that were primarily enriched in mammary gland morphogenesis and hormone levels related pathways, suggesting that mammary gland morphogenesis might play a unique role in TNBC patients differing from other breast cancer types. Further survival analysis revealed that nine genes (FSIP1, ADCY5, FSD1, HMSD, CMTM5, AFF3, CYP2A7, ATP1A2, and C11orf86) were significantly associated with the prognosis of TNBC patients, while three of them (ADCY5, CYP2A7, and ATP1A2) were involved in the hormone-related pathways. These findings indicated the vital role of the hormone-related genes in TNBC tumorigenesis and may provide some independent prognostic markers as well as novel therapeutic targets for TNBC.     

Overexpression of long non-coding RNA CCAT1 is a novel biomarker of poor prognosis in patients with breast cancer

Introduction: Recent studies have demonstrated that lncRNA CCAT1 was increased in many types of cancers and was involved in various cellular processes related to carcinogenesis. However, the clinical significance and prognostic value of lncRNA CCAT1 in breast cancer (BC) haven’t been investigated. Methods: Expression levels of lncRNA CCAT1 in 92 pairs of BC cancer tissues and adjacent normal tissues were detected by quantitative real-time PCR. In order to determine its prognostic value, overall survival and progression-free survival were evaluated using the Kaplan-Meier method, and multivariate analysis was performed using the Cox proportional hazard analysis. Results: Expression levels of lncRNA CCAT1 in BC tissues were significantly higher than those in adjacent normal tissues. High expression of lncRNA CCAT1 was associated with differentiation grade, TNM stage, and lymph node metastases. Kaplan-Meier analysis with the log-rank test indicated that high expression of lncRNA CCAT1 had a decreased overall survival and progression-free survival. Multivariable analysis was further identified high expression of lncRNA CCAT1 as an independent prognosis factor for overall survival and progression-free survival. Conclusions: Our findings provided that the expression of lncRNA CCAT1 was up-regulated in BC and associated with overall survival as well as progression-free survival, suggesting that lncRNA CCAT1 could be a potential prognostic biomarker for BC progression.     

限制片段差异显示PCR分析金属基质蛋白酶家族在乳腺癌表达的差异

目的：建立乳腺癌差异表达谱,以筛选乳腺癌相关候选基因;比较金属基质蛋白酶家族(matrix metalloproteinases,MMPs)在乳腺癌中的表达差异。方法：采集乳腺癌组织、转移淋巴结及正常乳腺组织,用限制片段差异显示PCR技术(Restriction fragments differential display PCR,RFDD-PCR)建立表达谱。以电泳和荧光扫描成像对表达谱片段进行分离、显示。筛选分析MMPs及其相关基因的表达差异。结果：共获得5407个基因片段,差异片段占30％以上;共显示13个MMPs基因及其相关基因金属蛋白酶组织抑制因子和转化生长因子β。其中除MMP20外,其余均有量或质的差异。结论：有多个MMPs基因参与了乳腺癌的发生、发展过程,其中MMP2、MMP10、MMP11、MMP14、MMP15、MMP28基因的开启可能为肿瘤进程的早期事件。     

基因芯片筛选Bardct-Bicdl综合征患者外周血差异表达基因

目的 采用基因表达谱芯片研究Bardet-Biedl综合征(Bardet-Biedl syndrome,BBS)患者外周血单个核细胞差异基因表达谱.方法 抽取3例BBS先证者及4名年龄、性别匹配的健康对照者静脉血,分离外周血单个核细胞,用Trizol一步法提取总RNA并进行扩增和标记,然后与Affymetrix U133 Plus 2.0芯片进行杂交,扫描芯片并将图像信号转化为数字信号,GCOS1.4软件读取、处理数据.将患者与正常对照基因表达谱进行比较,最后以表达差异≥2倍(P＜0.05)为标准来确定差异表达基因.结果 BBS患者与正常对照相比,有30个基因表达有显著差异,包括15个上调基因及15个下调基因.通过GO分析,有12个基因与信号传导及细胞周期有关.结论 筛选到的差异基因与纤毛的功能或结构相关性及在BBS发生及发展中的作用有待研究.     

心房颤动患者心房组织中差异表达miRNA的初步研究

目的：应用基因芯片技术研究心房颤动（房颤）患者心房组织中miRNA的表达谱,分析差异表达的miRNA,为进一步研究miRNA在房颤发生、发展中的作用奠定基础。方法：采用miRNA基因芯片技术检测房颤患者和非房颤患者心房组织样本中miRNA的表达水平;采用实时定量PcR（quantitativereal,timePCR,qRT．PCR）对部分差异表达的miRNA进行验证。结果：MiRNA基因芯片分析结果显示,与非房颤组织相比,房颤组织中差异表达的miRNA共有26个,其中16AmiRNA表达上调,10个miRNA表达下调。qRT．PCR验证结果与芯片结果相一致。结论：MiRNA在房颤患者心房组织中存在差异性表达。     

Comparative expressed sequence hybridization reveals differential gene expression in morphological breast cancer subtypes

In this study, comparative expressed sequence hybridization (CESH) has been used to compare gene expression patterns in three morphologically different breast cancer subtypes: classic-type invasive lobular carcinoma (ILC), poorly differentiated ERBB2-negative invasive ductal carcinoma-not otherwise specified (IDC-NOS), and poorly differentiated ERBB2-positive IDC-NOS. CESH allows global detection of chromosomal regions with differential gene expression in a way similar to that of comparative genomic hybridization (CGH). Eight cases of each breast cancer subtype were included in the study. For each subtype, two pools of four cases each were constructed. CESH was used to compare both pools within the same morphological subtype, followed by a comparison of pools belonging to different subtypes. This revealed three chromosomal regions that were differentially expressed in ductal and lobular carcinomas, including relative overexpression at 8q13-q23 and 16q22, and relative underexpression at 8p21-p22. In addition, an expression signature characterized by relative overexpression at 3q24-q26.3, 14q23-31, 17q12, and 20q12-13 was identified for ERBB2-positive IDC-NOS. In summary, CESH analysis highlights chromosomal regions of differential gene expression that are associated with morphologically defined breast cancer subtypes and suggests that regions on chromosome 8 are of interest in the discrimination between ductal and lobular carcinomas. In addition, using CESH, it was possible to identify an ERBB2 expression signature, comprising four chromosomal regions with potential significance in the aggressive behaviour of ERBB2-positive IDC-NOS.     

luminal A型乳腺癌组织中长链非编码RNA ENST00000460164的表达及对细胞周期的影响

目的明确lncRNA-ENST00000460164在人luminal A型乳腺癌组织中的表达及作用。方法用RT-q PCR检测ENST00000460164在17例配对的luminal A型乳腺癌及癌旁组织中的表达。将pll3.7-ENST00000460164-shRNA及pll3.7空载体转染MCF-7细胞,用流式细胞计量技术和Western blot检测转染后MCF-7细胞周期及其关键蛋白cyclin D1和P16INK4A的表达。结果长链非编码RNA ENST00000460164在luminal A型乳腺癌组织中呈高表达趋势;在MCF-7细胞中敲低ENST0000460164后,G1期明显延长,S期明显缩短(P0.05);G1期关键调控蛋白cyclin D1表达水平明显降低,P16INK4A表达明显上调(P0.05)。结论 ENST00000460164在luminal A型乳腺癌组织中呈高表达状态;ENST00000460164可能通过调控cyclin D1或者P16INK4A的表达改变细胞周期。