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1.
背景:骨关节炎是多因素介导的复杂疾病,发病机制尚待挖掘,随着基因层面的不断深入研究,非编码核糖核酸调控作用显现并得以研究。通过检测软骨细胞退变miRNA表达谱变化有助于更好地理解骨软骨细胞退变的分子机制,并为骨关节炎的诊断和治疗开辟新的途径。目的:探讨基质细胞衍生因子1刺激骨关节炎软骨细胞后miRNA表达谱变化,为基因层面延缓关节软骨退变提供实验基础。方法:对10例膝关节骨关节炎患者于全膝关节置换手术过程中截骨后残留的软骨组织进行软骨细胞培养,随机分为实验组与对照组,两组细胞培养基为含体积分数10%胎牛血清及青链霉素双抗的高糖DMEM培养基。实验组培养基中另外加入100μg/L基质细胞衍生因子1,对照组不做任何处理。两组软骨细胞培养48 h后,进行下一步实验用于miRNA芯片筛选和实时定量PCR验证。软骨组织标本取材前皆告知患者并征得同意,该研究符合《医疗机构管理条例》相关要求,获得昆明医科大学第一附属医院伦理委员会批准。结果与结论:miRNA基因芯片初筛共有84个miRNAs发生变化,其中70个miRNAs上调,14个miRNAs下调。通过基因芯片筛选差异变化的miRNA,对变化显著的7个miRNA(miR-146a-5p、miR-124-3p、miR-130a-3p、miR-185-5p、miR-221-3p、miR-126-3p)进行qRT-PCR实验验证,其中miR-146a-5p、miR-124-3p及miR-126-3p的qRT-PCR结果与基因芯片结果一致。结果表明,基质细胞衍生因子1刺激骨关节炎软骨细胞后循环miRNA表达谱出现明显变化,miR-146a-5p、miR-124-3p及miR-126-3p可能与基质细胞衍生因子1刺激骨关节炎SDF-1/CXCR4信号通路反应有关。  相似文献   

2.
张岚  刘法昱 《解剖科学进展》2020,26(2):143-145,149
目的分别比较4例HNSCC患者癌和癌旁组织的环状RNA、微小RNA和mRNA芯片表达谱,构建hsa_circRNA_103580/hsa-miR-9-5P/PRDM15相关的ceRNA网络,探讨其表达趋势。方法利用芯片筛选HNSCC组织中差异表达的circRNA、miRNA和mRNA基因,根据miRwalk3.0在线软件预测的结果,构建HNSCC中的circRNA/miRNA/mRNA-相关的ceRNA网络。采用qRT-PCR方法检测HNSCC组织中hsa_circRNA_103580/hsamiR-9-5P/PRDM15基因的表达。结果芯片技术检测4例HNSCC患者癌和癌旁组织中差异表达的circRNA、miRNA及mRNA基因,获得显著差异的环状RNAs转录本311个,microRNAs转录本52个及circRNA433个。结合以上数据,构建了一个包含48种circRNAs,21种miRNAs和79种mRNAs的ceRNA网络并验证has-circRNA_103580、has-miR-9-5p与PRDM15的基因表达水平与芯片中表达趋势一致。结论 hsa_circRNA_103580/hsa-miR-9-5P/PRDM15轴可能是HNSCC预后和治疗新的靶点。  相似文献   

3.
目的通过TCGA数据库中浆液性卵巢癌(serous ovarian carcinoma,SOC)miRNA芯片表达谱数据分析,寻找与SOC预后显著相关的miRNAs,为后续研究提供可靠依据。方法下载TCGA数据库中SOC miRNAs表达数据及临床数据,应用SPSS20. 0软件进行数据合并及标准化处理,首先运用Spearman秩相关分析筛选miRNAs表达数据,再对筛选结果进行单因素Cox分析。结果 Spearman秩相关分析筛选出20个预后相关的miRNAs,再经单因素Cox分析筛出9个与SOC患者预后密切相关的miRNAs,经Kaplan-Meier生存曲线和ROC曲线分析验证,这些miRNAs的差异表达均与SOC预后相关。结论经Spearman秩相关分析和单因素Cox分析筛选出SOC预后相关的miRNAs的可信度较高,这些miRNAs可作为评价患者预后的新指标。  相似文献   

4.
 目的:筛选并分析喉癌组织与周围正常喉黏膜的微小RNA(microRNAs,miRNAs) 之间的表达谱差异,为进一步研究miRNA与喉癌发生、发展的关系提供线索。方法:收集喉癌组织和癌旁正常喉黏膜标本共42对,随机选取10对标本进行miRNA微阵列基因芯片分析, 另选取32对标本进行实时荧光定量PCR (qRT-PCR)验证,获得喉癌组织中的miRNA差异表达谱。应用MTT法和克隆形成实验检测miR-125a-5p对喉癌Hep2细胞增殖的影响。结果:喉癌组织中的let-7f-5p、miR-10a-5p、miR-125a-5p、miR-144-3p、miR-195-5p、miR-203等6个miRNA在基因芯片以及qRT-PCR中表达均显著下调。与对照组相比,转染miR-125a-mimics组的喉癌Hep2细胞增殖能力受到抑制,而转染miR-125a-inhibitor组Hep2细胞增殖能力增强。结论:基因芯片与qRT-PCR结果一致;喉癌与正常喉黏膜之间存在明显的miRNA差异表达,这些miRNA的差异性表达可能与喉癌的发病、侵袭等相关。miR-125a可以抑制喉癌Hep2细胞的增殖,可能作为喉癌生物治疗的新靶点。  相似文献   

5.
目的:应用基因芯片技术筛选结肠癌耐药相关微小RNAs (miRNAs),探究miRNAs对化疗耐药的调控机制。方法:采用基因芯片技术分析结肠癌细胞系HCT8及其耐长春新碱细胞系HCT8/v中miRNAs的表达差异,对部分差异表达的miRNAs应用RT-qPCR进行验证,对表达差异显著的miRNAs进行靶基因预测,利用Gene Ontology (GO)和京都基因与基因组百科全书(KEGG)数据库对预测到的靶基因进行生物信息学分析。结果:筛选出342个差异表达miRNAs,其中190个表达上调,152个表达下调。RT-qPCR验证结果示miR-125-5p、miR-181c-5p和miR-153-3的表达情况和芯片检测结果一致;miR-130a-3p和miR-149-3p的表达与芯片检测结果不一致。GO分析结果显示,耐药相关基因主要富集的旁路是RNA聚合酶II调控区序列特异性DNA结合旁路,主要通过正向调节发挥作用,位置主要是在细胞内有界细胞器上。KEGG分析结果显示,耐药相关基因最为富集的是轴突导向通路、胰岛素信号通路及磷脂酶D信号通路。结论:miRNAs与结肠癌化疗耐药密切相关。对这些miRNAs的研究能使我们对结肠癌的化疗耐药机制有更深入的理解,并为逆转化疗耐药提供新的思路。  相似文献   

6.
目的检测卵巢癌荷瘤小鼠癌组织miR-21-5p、miR-155-5p、miR-218-5p、miR-222-3p、miR-494-3p的水平和外周血和脾脏中髓源性抑制细胞(MDSC)的数量,并分析它们的相关性。方法 12例卵巢癌荷瘤小鼠的癌组织和癌旁组织,实时荧光定量PCR检测癌组织和癌旁组织中miR-21a-5p、miR-155a-5p、miR-218-5p、miR-222-3p、miR-494-3p的水平,流式细胞术检测上述荷瘤小鼠中外周血和脾脏中MDSC的表达。采用Spearman法分析MDSC和microRNA的相关性。结果与正常小鼠相比,荷瘤小鼠外周血和脾脏中的MDSC明显增加。miR-21a-5p、miR-218-5p和miR-222-3p在癌组织中含量高于癌旁组织,相反miR-155a-5p和miR-494-3p在癌组织中的表达低于癌旁组织。miR-222-3p与MDSC的数量无相关性,miR-494-3p的水平与MDSC的数量负相关,miR-21a-5p、miR-155a-5p、miR-218-5p在癌组织与癌旁组织的表达差值与MDSC的数量呈正相关。结论卵巢癌荷瘤小鼠癌组织中miR-21a-5p、miR-155a-5p、miR-218-5p、miR-494-3p水平和外周血及脾脏中MDSC的数量有一定的相关性。  相似文献   

7.
目的筛选和验证靶向调控c-SKI并与纤维化相关的microRNA(miRNA)。方法生物信息学方法预测并结合文献报道,筛选出靶向c-SKI的候选miRNAs,RT-qPCR检测人心肌成纤维细胞(HCFBs)中候选miRNAs和c-SKI的表达,筛选出抑制作用最显著的miRNA;构建c-SKI-3′-UTR野生型(c-SKI-wt)和突变型(c-SKI-mut)载体,分别与miR-155a-5p/miR-17a-5p的模拟物、抑制剂及对照在人胚肾上皮细胞(HEK293T)中共转染,双萤光素酶报告系统检测各组荧光素酶活性;接着,分别将miR-155a/miR-17a-5p mimics和inhibitor转染至人心肌成纤维细胞(HCFBs),Western blot检测各组细胞c-SKI的表达。结果 1)经筛选miR-155a-5p和miR-17a-5p对c-SKI的抑制作用最明显(P<0.01);2)与NC组相比,miR-155a-5p/miR-17a-5p mimics组萤光素酶活性均显著下降(P<0.05),miR-155a-5p/miR-17a-5p inhibitor组萤光素酶活性均明显增强(P<0.05);3)与NC组相比,miR-155a-5p/miR-17a-5p mimics组中c-SKI蛋白表达显著下调,miR-155a-5p/miR-17a-5p inhibitor组中c-SKI的表达显著上调(P<0.01)。结论 miR-155a-5p和miR-17a-5p可分别靶向结合c-SKI的3′-UTR,在HCFBs中负性调控c-SKI的表达。  相似文献   

8.
目的探索永久性房颤(p AF)发病相关的关键miRNAs及其调控的靶基因。方法联合应用表达谱芯片和miRNA芯片分析p AF患者(n=7)和健康成人(n=4)的左房组织,筛选p AF相关差异表达的miRNAs,进行靶基因预测后,与表达谱芯片的筛选结果进行负相关分析后的基因集合进行显著性功能分析(GO-analysis);利用miRNA与靶基因之间的靶向调控关系,构建差异miRNA与交集靶基因的调控网络(miRNA-gene-network),得到网络中起核心调控作用的miRNA和被调控的关键靶基因;采用RT-q PCR方法检验另一组p AF患者(n=5)和健康成人(n=4)的左房组织标本。结果表达谱基因芯片发现610个mRNA有显著性改变(fold change2,P0.05),miRNA-靶基因调节网络发现与p AF显著相关的20个miRNAs和107个靶基因,相关度最高的是miR-144、miR-1284、miR-1827、miR-1、miR-3613-3p和miR-101;其调控的重要靶基因包括CACNB2、EFNB1、PTEN、TAOK1、RUNX1和TPM3等;RT-q PCR验证结果显示这些miRNAs和靶基因密切相关。结论通过表达谱基因芯片与miRNAs芯片联合分析p AF左房组织标本,构建miRNA调控网络,发现p AF重要的miRNA-靶基因调控及功能,结果更加准确。  相似文献   

9.
目的 探讨miR-99a-5p通过靶点SLC44A1对肺腺癌(LUAD)细胞系增殖的抑制作用及可能机制.方法 从基因表达综合数据库(GEO)中检索并筛选GPL18058的microRNA(miRNA)表达阵列,筛选出肺癌与正常组织差异表达的miR-99a-5p后,通过Target Scan Human数据库预测其下游靶点SLC44A1,并利用UALCAN分析SLC44A1在肺癌组织中的表达情况和LUAD预后关系.利用实时荧光定量PCR(qRT-PCR)检测肺腺癌细胞系miR-99a-5p和SLC44A1的表达水平.在A549细胞中转染miR-99a-5p模拟物,通过CCK-8试验、克隆形成实验检测A549的增殖情况;qRT-PCR和Western blot检测SLC44A1的表达.在A549细胞中过表达SLC44A1,评估其对肺癌调节中的miR-99a-5p的选择性作用.结果 在LUAD组织和细胞系中,miR-99a-5p的表达异常下调,过度异位表达的miR-99a-5p抑制A549细胞的增殖(P<0.05).在LUAD组织和细胞系的基因和蛋白质水平上,miR-99a-5p负性调控SLC44A1(P<0.05).SLC44A1过度表达有效地逆转了miR-99a-5p在体外对A549细胞增殖的抑制作用.SLC44A1的低表达有利于患者生存率的提高(P<0.05).结论 miR-99a-5p抑制LUAD中A549细胞系的增殖,并通过负性调控SLC44A1发挥作用.  相似文献   

10.
目的基于公用数据库,寻找可作为乳腺癌生物标记物的miRNAs。方法利用人类肿瘤基因组(TCGA)数据库下载乳腺癌相关数据集,分析肿瘤组织与正常组织之间miRNAs的差异表达。Cox单因素回归分析与不良预后相关的miRNAs;对差异表达中上调的有意义的miRNAs进行Cox多因素回归分析,建立风险评估模型。结果与正常组织比较,乳腺癌组织中有370个差异表达miRNAs,其中108个miRNAs表达下调,262个miRNAs表达上调。20个miRNAs的高表达与预后不良相关。进一步建立Cox回归模型筛选出10个miRNAs组合(包括hsa-mir-148b、hsa-mir-449c、hsa-mir-106a、hsa-mir-181d、hsa-mir-9-3、hsa-mir-549a、hsa-mir-556、hsa-mir-618、hsa-mir-466、hsa-mir-135a-1)预测乳腺癌不良预后。结论筛选的10个miRNAs组合(ten-miRNAs)的高评分可成为预测乳腺癌不良预后的生物标记物。  相似文献   

11.
Accumulating evidence has demonstrated that aberrantly expressed miRNAs in cancer tissues regulated various cellular processes related to carcinogenesis. The present study aimed to identify the differentially expressed miRNAs between esophageal squamous cell cancer (ESCC) and adjacent normal esophageal tissue (ANET). In our present study, we identified 129 differentially expressed miRNAs between ESCC and ANET by analyzing high-throughput miRNA data downloaded from TCGA database. After investigating the prognostic value of the 129 differential expressed miRNAs, eight miRNAs were found to be associated with prognosis of patients with ESCC. The clinical significance and bio-function of miR-375 was further examined. We performed Gene Set Enrichment Analysis (GSEA) to identify the top three gene sets that significantly altered between the patients with miR-375 low expression and high expression. In order to explore the mechanism of the development and progression of ESCC, the role of miR-375 in ESCC and its four candidate target genes was examined. At last, we performed a meta-analysis to verify the prognostic value of miR-375 in ESCC. In conclusion, our findings suggest that miR-375 serves as a promising independent prognostic factor for ESCC and function as a tumor suppressor.  相似文献   

12.

Background

The role of miR-99a-3p in Head and neck squamous cell carcinoma (HNSCC) has not been reported. Therefore, in this study, we examined the expression level and its molecular mechanisms of miR-99a-3p in HNSCC.Materials and methods: MiR-99a-3p-related miRNA-chip and miRNA-sequencing data were collected. We then carried out meta-analyses to pool the standard mean difference (SMD) value and generate a summarized receiver operating characteristic (sROC) curve. MiR-99a-3p mimic was transfected into FaDu cells and those genes influenced by miR-99a-3p were gathered. The target genes were also predicted from 12 tools through miRwalk2.0, and combined with differentially expressed genes in HNSCC from the The Cancer Genome Atlas and Genotype-Tissue Expression sequencing databases. FunRich and DAVID were used for the pathway signaling analyses for the potential targets of miR-99a-3p in HNSCC.

Results

The SMD was -0.30 (95% CI: -0.51, -0.08) in the fixed-effect model and -0.28 (95% CI: -0.67, 0.10) in the random-effect model (I2?=?60%), indicating a reduced expression level of miR-99a-3p in HNSCC tissues based on 1167 cases. In the sROC curve, the area under the curve (AUC) was 0.77 (95% CI: 0.73, 0.81). The 251 potential targets of miR-99a-3p were enriched in several pathways related to cancer, with the “Pathways in cancer” standing at the top. vascular endothelial growth factor A was selected as an example with up-regulated trend in HNSCC tissues.

Conclusion

MiR-99a-3p exhibits a significant lower expression status in HNSCC, and this reduced or deletion status promotes the malignant progression of HNSCC. However, its molecular mechanism is still unclear and requires further investigation.  相似文献   

13.
BACKGROUND: Liver regeneration is the key factor influencing the prognosis of living donor liver transplantation. There has not been the research on special miRNA of liver regeneration after living donor liver transplantation. OBJECTIVE: To analyze the variation of miRNAs expression profile after rat reduced-size liver transplantation at certain time point, select and verify target miRNA which can provide targeting intervention strategies in liver regeneration after rat reduced-size liver transplantation and provide theoretical evidence for liver regeneration after living donor liver transplantation. METHODS: The reduced-size liver transplantation models were established. miRNAs microarray was used to detect miRNA expression. In differentially expressed microRNAs, real-time quantitative PCR was utilized to detect target miRNAs. The credibility of miRNAs microarray results was verified. RESULTS AND CONCLUSION: Compared with rat liver tissue in the sham operation group, 11 miRNAs up-regulated in reduced-size liver transplantation, including let-7b-5p, let-7c-5p, miR-101a-3p, miR-103-3p, miR-130a-3p, miR-142-5p, miR-186-5p, miR-199a-3p, miR-21-5p, 221-3p and miR-34a-5p. Four miRNAs were down-regulated, including miR-26b-5p, miR-150-5p, miR-19a-3p and rno-miR-146-5p. PCR test further verified that miR-221-3p and miR-199a-3p expression changes approximated the chip results at 24, 48 hours and 1 week, indicating that results of miRNA microarray were believable. These results verified that it exists variation of miRNAs expression profile after rat reduced-size liver transplantation, which picked out and verified the target miRNAs.     相似文献   

14.
Small, noncoding microRNAs (miRNAs) have been shown to be abnormally expressed in every tumor type examined. We used comparisons of global miRNA expression profiles of head and neck squamous cell carcinoma (HNSCC) samples and adjacent normal tissue to rank those miRNAs that were most significantly altered in our patient population. Rank Consistency Score analysis revealed miR-375 to have the most significantly lowered miRNA levels in tumors relative to matched adjacent nonmalignant tissue from the same patient among 736 miRNAs that were evaluated. This result has been previously observed by other groups; however, we extend this finding with the unique observation that low miR-375 expression levels correlate significantly with cancer survival and distant metastasis. In a study of 123 primary HNSCC patients using multivariable Cox proportional hazard ratios (HR) and 95% confidence intervals (CI), both death from disease (HR: 12.8, 95% CI: 3 to 49) and incidence of distant metastasis (HR: 8.7, 95% CI: 2 to 31) correlated with lower expression levels of miR-375 regardless of the site or stage of the tumor. In addition, we found that oral cavity tumor cell lines (eg, UMSCC1 and UMSCC47) overexpressing miR-375 were significantly less invasive in vitro than their matched empty vector controls. We conclude that miR-375 represents a potential prognostic marker of poor outcome and metastasis in HNSCC and that it may function by suppressing the tumor's invasive properties.  相似文献   

15.
目的 筛选并鉴定在食管鳞癌(ESCC)及癌旁组织中miRNAs的差异表达,为进一步阐明其在ESCC发病机制中的作用奠定基础。方法 选取在邯郸市第一医院行食管癌切除术患者新鲜的鳞癌组织及癌旁正常组织,各3例,抽提总RNA,利用miRNAs芯片筛选其中差异表达的miRNAs,并对miR-106b-3p通过进一步RT-qPCR 技术进行验证。结果 miRNAs芯片从配对组织中共筛检出62个差异表达的miRNAs,其中41个miRNAs表达上调,21个miRNAs表达下调,鳞癌组织与癌旁正常组织中差异表达的miRNAs比较,差异有统计学意义(P<0.05)。miR-106b-3p在ESCC组织中的表达高于癌旁组织,差异有统计学意义(P<0.05),与芯片结果一致。结论 食管鳞癌组织中miRNAs的差异表达为进一步研究miRNAs在ESCC发病中的作用奠定了基础;miR-106b-3p的高表达可能与ESCC的发生、发展有关。  相似文献   

16.
BackgroundRenal cell carcinoma (RCC) accounts for 3 % of cancer patients. Early detection influences the therapeutic strategy and significantly improves patients’ survival rates. Stable existing circulating miRNAs could be a promising diagnostic biomarker.MethodsPreviously our team demonstrated the anti-tumor effect of miR-20b-5p, miR-30a-5p and miR-196a-5p in RCC tissue and cell lines. Here, based on 110 RCC patients and 110 health control, we investigated serum expression of these three miRNAs in the testing set and the validation set separately by using quantitative real-time PCR. A three-miRNA panel with high diagnostic efficiency was constructed. Correlations between these miRNAs and clinical parameters were investigated. Additionally, the TCGA dataset and bioinformatic analysis are used for the functional exploration of these miRNAs.ResultsSerum expression levels of miR-20b-5p, miR-30a-5p were significantly reduced in RCC patients, while miR-196a-5p expression level was up-regulated (p < 0.001). miR-20b-5p, miR-30a-5p and miR-196a-5p had moderate diagnostic ability for RCC (AUC = 0.807, 0.766 and 0.719 in the testing set, respectively). The AUC of the three-miRNA panel was 0.949 in the testing set and 0.938 in the validation set. Specifically, the serum expression level of miR-196a-5p was significantly down-regulated in RCC patients with higher Fuhrman grade (p = 0.051). TCGA dataset analysis showed that the three-miRNA panel probably participated in RCC by targeting ITGA4 and NRP2.ConclusionThe three-miRNA panel could serve as a promising non-invasive biomarker for RCC detection.  相似文献   

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目的 建立以甲醛固定石蜡包埋组织为材料、基于基因芯片技术的microRNA(miRNA)表达谱的分析方法 ;筛选与喉鳞状细胞癌(简称喉癌)生物学特征密切相关的差异表达miRNA.方法 从喉癌甲醛固定石蜡包埋组织中制备总RNA,经质量鉴定后进行荧光标记.采用Agilent公司的容纳723条人类miRNA探针的基因芯片完成杂交实验,以获得喉癌的miRNA表达谱.以GeneSpring GX和R-Project软件处理分析基因芯片实验数据,筛选与喉癌转移相关的差异表达miRNA.结果 从24例甲醛固定石蜡包埋组织标本中获得了符合基因芯片实验质量标准的RNA样品,并完成了基因芯片杂交及数据分析.从中共鉴定到319个miRNA,有96个miRNA在24例喉癌中均有表达,其中与淋巴结转移密切相关的(检错率<0.05)差异表达miRNA有5个,分别为miR-23a* 、miR-28-5p、miR-15a、miR-16和miR-425.结论 甲醛固定石蜡包埋组织可以提供符合基因芯片分析质量要求的miRNA,是研究miRNA的重要样品资源.从喉癌的miRNA表达谱中筛选出的转移相关差异表达miRNA(miR-23a*、miR-28-5p、miR-15a、miR-16和miR-425)有可能成为评估喉癌转移风险的新型分子标志.  相似文献   

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目的 建立以甲醛固定石蜡包埋组织为材料、基于基因芯片技术的microRNA(miRNA)表达谱的分析方法 ;筛选与喉鳞状细胞癌(简称喉癌)生物学特征密切相关的差异表达miRNA.方法 从喉癌甲醛固定石蜡包埋组织中制备总RNA,经质量鉴定后进行荧光标记.采用Agilent公司的容纳723条人类miRNA探针的基因芯片完成杂交实验,以获得喉癌的miRNA表达谱.以GeneSpring GX和R-Project软件处理分析基因芯片实验数据,筛选与喉癌转移相关的差异表达miRNA.结果 从24例甲醛固定石蜡包埋组织标本中获得了符合基因芯片实验质量标准的RNA样品,并完成了基因芯片杂交及数据分析.从中共鉴定到319个miRNA,有96个miRNA在24例喉癌中均有表达,其中与淋巴结转移密切相关的(检错率<0.05)差异表达miRNA有5个,分别为miR-23a* 、miR-28-5p、miR-15a、miR-16和miR-425.结论 甲醛固定石蜡包埋组织可以提供符合基因芯片分析质量要求的miRNA,是研究miRNA的重要样品资源.从喉癌的miRNA表达谱中筛选出的转移相关差异表达miRNA(miR-23a*、miR-28-5p、miR-15a、miR-16和miR-425)有可能成为评估喉癌转移风险的新型分子标志.  相似文献   

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