肝细胞癌根治术后早期复发相关microRNAs的筛选

目的 筛选与肝细胞癌（hepatocellular carcinoma,HCC）患者根治术后早期复发相关的microRNAs。方法 收集10例HCC患者肝脏组织标本,其中早期复发组5例,非早期复发组5例,以及正常肝脏组织标本4例。利用microRNA基因芯片技术检测3组肝脏组织标本中microRNAs的表达谱,筛选出差异表达的microRNAs。结果 早期复发组与正常组相比,miR-21-3p、miR-21-5p、miR-222-3p、miR-3182、miR-3651、miR-3654、miR-4451、miR-4633-5p、miR-720的表达均上调,miR-4525的表达下调（P〈0.05）;非早期复发组与正常组相比,miR-106b-5p、miR-4451、miR-3651、miR-93-5p的表达均上调,miR-451a、miR-3692-5p的表达下调（P〈0.05）。结论 microRNAs的表达与HCC患者术后早期复发有关,检测患者肝脏组织中microRNAs的表达水平可能有助于监测HCC术后早期复发。     

miR-886-5p抑制肝细胞癌细胞侵袭与迁移

目的:探讨miR-886-5p在肝细胞癌(hepatocellular carcinoma,HCC)中的表达水平及其与临床病理特征的关系,并分析miR-886-5p对HCC细胞侵袭与迁移能力的影响.方法:应用Real-time PCR技术检测HCC组织和细胞系中miR-886-5p的表达水平;分析miR-886-5p的表达水平与HCC临床病理特征的关系;应用Transwell实验探究miR-886-5p对HCC细胞侵袭与迁移能力的影响.结果:miR-886-5p在HCC组织中的表达水平较癌旁组织明显降低(P＜0.05),miR-886-5p在HCC细胞系中(Hep3B、MHCC-97L、HepG2、SMMC-7721)的表达水平较正常肝细胞LO2显著下调(P＜0.05);miR-886-5p的表达水平与肿瘤数目、静脉侵犯、Edmondson病理分级及TNM分期显著相关(P＜0.05);Transwell小室实验表明miR-886-5p能够显著抑制HCC细胞的侵袭与迁移能力.结论:miR-886-5p在HCC中表达下调,其能够抑制HCC细胞的侵袭与迁移.     

MiR 142 5p通过靶向IGF2BP3调控多柔比星诱导的肝癌细胞凋亡

目的:探讨miR-142-5p对多柔比星诱导的原发性肝细胞癌(hepatocellular carcinoma,HCC)细胞凋亡的影响及其作用机制.方法:收集广西医科大学附属肿瘤医院88例HCC患者手术切除的癌组织及癌旁组织(距癌灶组织边缘2～5cm)标本.采用实时荧光定量PCR检测人HCC组织和癌旁组织、人正常肝细胞以及HCC细胞系中miR-142-5p的表达量.向HCC细胞SMMC-7721中转染miR-142-5p mimics,流式细胞术检测过表达miR-142-5p后SMMC-7721细胞在多柔比星(doxorubicin)(1 μg/ml)诱导下凋亡的变化;生物信息学方法预测miR-142-5p可靶向结合胰岛素样生长因子2 mRNA结合蛋白3(in-sulin-like growth factor 2 mRNA-binding protein 3,IGF2BP3)基因,并采用荧光素酶报告基因实验进行验证.采用实时荧光定量PCR及Western blotting检测过表达miR-142-5p的SMMC-7721细胞中IGF2BP3的mRNA及蛋白表达情况.结果:与癌旁组织和正常肝细胞相比,HCC组织(-6.91±2.61vs-11.59±2.59,P＜0.01)和多种HCC细胞系中miR-142-5p呈明显低表达(均P＜0.01);过表达miR-142-5p可显著促进多柔比星诱导的HCC细胞SMMC-7721的凋亡[(49.40±3.47)％ vs (19.50±1.74)％,P＜0.01];过表达miR-142-5p可明显降低HCC细胞中IGF2BP3的mRNA及蛋白表达水平(P＜0.01),敲减IGF2BP3表达可进一步促进多柔比星诱导的SMMC-7721细胞的凋亡(P＜0.01).荧光素酶报告基因实验结果显示,miR-142-5p能够抑制IGF2BP3的3'UTR荧光素酶报告基因的活性.结论:miR-142-5p在HCC组织标本和体外培养细胞系中的表达水平均显著降低,转染miR-142-5p mimics后能够促进多柔比星诱导的HCC细胞的凋亡,其机制可能与miR-142-5p靶向作用IGF2 BP3从而促进HCC细胞凋亡有关.     

miR-369-5p抑制肝细胞癌细胞增殖、迁移及侵袭

目的:探讨miR-369-5p在肝细胞癌(hepatocellular carcinoma,HCC)中的表达情况、临床意义,观察miR-369-5 p对HCC细胞增殖、迁移及侵袭的影响及其作用机制.方法:实时定量PCR检测miR-369-5p在HCC组织与相应癌旁组织、正常肝细胞(L02)与HCC细胞系中的表达情况;C...     

miR-141-3p在胃癌组织和患者血浆中的表达及其临床意义

目的:检测miR-141-3p在胃癌组织及患者血浆中的表达水平,探讨其表达水平与患者病理特征和预后的关系.方法:收集河北医科大学第四医院普外科2012年5月至2013年1月胃癌根治性手术切除的组织标本及术前外周静脉血标本44例,每例标本采集肿瘤组织(非坏死部分)和配对的癌旁组织.采用实时荧光定量PCR法检测miR-141-3p在胃癌组织、癌旁组织、血浆及健康志愿者血浆(25例)的表达情况,分析miR-141-3p的表达水平与胃癌患者DFS及临床病理特征的关系.结果:miR-141-3p在胃癌组织中的表达明显降低,在血浆中的表达水平明显升高(均P＜0.01).在胃癌组织中miR-141-3p高表达组DFS明显高于低表达组(21.8 vs 10.3个月,P＜0.01),且miR-141-3p在癌组织中的表达水平与患者DFS呈正相关(P＜0.01).在血浆中miR-141-3p的高表达组DFS明显低于低表达组(9.1 vs 21.0个月,P＜0.01),且miR-141-3p血浆中的表达水平与患者DFS呈负相关(P＜0.01).miR-141-3p在胃癌患者血浆中表达水平与肿瘤TNM分期、淋巴结转移及脉管瘤栓相关(P＜0.01).结论:miR-141-3p在胃癌组织中表达显著降低且与患者预后呈正相关,其在患者血浆中表达显著升高且与患者预后呈负相关,表明miR-141-3p可作为胃癌早诊早治及患者临床预后判定的潜在标志物.     

miR-24-3p在胰腺癌中的表达及意义

目的:分析57例胰腺癌患者的癌组织与癌旁组织中的miR-24-3p的表达情况及临床意义。方法:收集57例胰腺癌初诊患者术后的癌组织及癌旁组织，用实时荧光定量PCR技术检测miR-24-3p在胰腺癌组织和相应癌旁组织中的表达情况，用χ2检验分析miR-24-3p的表达与胰腺癌患者临床病理指标之间的相关性；用Kaplan－Meier单因素分析计算累积生存率和Log-rank检验分析胰腺癌组织中miR-24-3p的表达量与胰腺癌患者术后预后的关系。结果:miR-24-3p在胰腺癌组织中相对高表达；miR-24-3p的表达与患者的年龄、性别、肿瘤分化程度、肿瘤位置和血管浸润无关（P＞0.05）,而与肿瘤的大小、TNM分期和淋巴结转移相关（P＜0.05）；miR-24-3p高表达组患者的3年累计生存率明显低于miR-24-3p低表达组患者（P＜0.05）。结论:miR-24-3p在胰腺癌组织中的表达明显高于其癌旁组织，提示其参与了胰腺癌的发生发展过程。     

miR-363-3p和miR-5100在非小细胞肺癌中的表达及其临床意义

目的:探讨小分子RNA(microRNA)miR-363-3p 和miR-5100在非小细胞肺癌中的表达及其临床意义.方法:利用荧光定量PCR的方法,检测肿瘤组织、癌旁组织及远离肿瘤的正常组织中致癌基因Myc的mRNA 和miR-363-3p、miR-5100的表达,然后分析miR-363-3p 和miR-5100与临床病理特征之间的相关性.结果:Myc在肿瘤组织中表达明显升高;miR-363-3p在肺癌组织中的表达明显低于正常组织(P<0.001),而在癌旁组织中的表达却远远高于正常组织(P<0.05);miR-5100在肺癌组织中的表达显著地高于正常组织(P<0.001),并且癌旁组织中的表达也高于正常组织(P<0.05).临床相关性分析显示,miR-363-3p的表达与淋巴结转移呈正相关(P<0.001),miR-5100的表达与临床分期也呈正相关性(P<0.05).结论:miR-363-3p 和miR-5100可能参与了非小细胞肺癌早期的发生和转移,并可能作为早期诊断和预后的分子标志物.     

miR-802在原发性肝细胞癌中的表达及意义

[目的]检测微小RNA(miR)-802在原发性肝细胞癌(hepatocellular carcinoma,HCC)组织与细胞中的表达水平,分析miR-802在HCC中的临床意义。[方法]采用qRT-PCR检测HCC癌组织与癌旁组织中mi R-802的表达水平,及Hep3B、HCCLM3与HL-7702细胞的表达水平。分析miR-802表达水平与HCC临床病理特征的关系。采用Kaplan-Meier方法计算生存曲线,以Log-rank检验评估miR-802表达水平与HCC预后的关系。通过Cox比例风险回归模型单因素和多因素分析评估HCC的预后指标。[结果] HCC癌组织中miR-802的表达水平显著低于癌旁组织(P<0.05),且Hep3B、HCCLM3细胞中mi R-802的表达水平亦显著低于HL-7702细胞(P<0.05)。Ⅲ+Ⅳ期癌组织中miR-802的表达水平低于Ⅰ+Ⅱ期癌组织(P<0.05),血管转移患者癌组织中miR-802的表达水平低于非血管转移患者癌组织(P<0.05)。mi R-802低表达组患者的5年生存率显著低于高表达组(P<0.05)。单因素分析显示,低表达mi R-802、高TNM分期及阳性血管转移是HCC患者预后不良的预测指标(P<0.05)。多因素分析显示,低表达miR-802是HCC患者预后不良的唯一独立预测指标(P<0.05)。[结论] HCC组织和细胞中miR-802表达下调,低表达miR-802是HCC患者预后不良的独立预测指标。     

MicroRNA-338-3p在结直肠癌中的表达及其临床意义

背景与目的:MicroRNA(miRNA,miR)是一类可负性调控基因表达的非编码小RNA,能通过与mRNA分子相互作用阻遏蛋白质翻译或导致RNA降解;且在肿瘤的发生、发展中扮演着重要的角色。本研究旨在探讨microRNA-338-3p(miR-338-3p)在结直肠癌与癌旁组织中差异表达状况及其与临床病理特征和预后之间的关系。方法:选取南方医院2008年9—11月间经手术切除的原发性结直肠癌标本40例,完善相关病理资料;常规抽提肿瘤及对照癌旁组织中总RNA,应用实时荧光定量PCR检测标本中miR-338-3p表达状况,并分析其与临床病理特征及预后的关系。结果:40例结直肠癌与癌旁组织相比,miR-338-3p表达明显下调(0.122 6±0.087 3 vs0.905 8±0.410 5),差异有统计学意义(P<0.01);miR-338-3p表达水平与肿瘤TNM分期(P=0.031)、浸润深度(P=0.001)以及分化程度(P=0.042)有关;miR-338-3p表达水平越低,其3年无进展生存率及总生存率越低。结论:MiR-338-3p可能作为"抑癌基因"参与了结直肠癌的发生、发展,并可作为结直肠癌预后判断的新的生物标志物。     

子宫颈鳞状细胞癌组织中LncRNA HCG11和miR-590-3p的表达及其与预后的关系

 目的 探讨LncRNA HCG11和miR-590-3p在子宫颈鳞状细胞癌组织及配对的癌旁组织中的表达情况,以及二者表达水平与患者临床资料和预后的相关性。方法 收集58例子宫颈鳞状细胞癌和配对的癌旁正常组织标本,运用qRT-PCR法检测58对子宫颈鳞状细胞癌组织及相应的癌旁组织中的LncRNA HCG11和miR-590-3p的表达,分析与宫颈鳞状细胞癌临床病理和预后的相关性,并评价其临床意义。结果 荧光定量PCR显示宫颈鳞状细胞癌组织中LncRNA HCG11的表达比癌旁组织明显下调（P<0.05）,miR-590-3p的表达比癌旁组织明显上调（P<0.05）;二者在核酸表达水平上呈负相关（r=-0.642, P=0.000）。 LncRNA HCG11和miR-590-3p的表达均与宫颈癌淋巴结转移、组织大小及临床分期之间存在显著的相关性,且与预后相关（P<0.05）。Cox回归多因素分析发现LncRNA HCG11和miR-590-3p分别可作为宫颈鳞状细胞癌患者独立的预后因子。结论 LncRNA HCG11和miR-590-3p可作为宫颈鳞状细胞癌独立的预后标志物和潜在的治疗靶点,LncRNA HCG11可能通过调控miR-590-3p的表达量从而发挥抑癌基因的作用。     

miRNA-152在肝癌组织中的表达及临床意义

目的 探讨微小RNA(miR)-152在肝癌组织中的表达及临床意义.方法 收集56例肝癌患者癌及癌旁组织标本,采用实时荧光定量聚合酶链反应技术(Real-time PCR)检测肝癌及癌旁组织标本中miR-152的表达水平,分析其与肝癌患者临床病理特征及预后的相关性.结果 miR-152在肝癌及癌旁组织中相对表达水平分别...     

MicroRNA expression profiles in serous ovarian carcinoma.

PURPOSE: Although microRNAs have recently been recognized as riboregulators of gene expression, little is known about microRNA expression profiles in serous ovarian carcinoma. We assessed the expression of microRNA and the association between microRNA expression and the prognosis of serous ovarian carcinoma. EXPERIMENTAL DESIGN: Twenty patients diagnosed with serous ovarian carcinoma and eight patients treated for benign uterine disease between December 2000 and September 2003 were enrolled in this study. The microRNA expression profiles were examined using DNA microarray and Northern blot analyses. RESULTS: Several microRNAs were differentially expressed in serous ovarian carcinoma compared with normal ovarian tissues, including miR-21, miR-125a, miR-125b, miR-100, miR-145, miR-16, and miR-99a, which were each differentially expressed in >16 patients. In addition, the expression levels of some microRNAs were correlated with the survival in patients with serous ovarian carcinoma. Higher expression of miR-200, miR-141, miR-18a, miR-93, and miR-429, and lower expression of let-7b, and miR-199a were significantly correlated with a poor prognosis (P < 0.05). CONCLUSION: Our results indicate that dysregulation of microRNAs is involved in ovarian carcinogenesis and associated with the prognosis of serous ovarian carcinoma.     

MIR-107, MIR-223-3P and MIR-21-5P Reveals Potential Biomarkers in Penile Cancer

Background: Inguinal lymph node involvement is the main prognostic factor in patients with penile cancer. However, there is a lack of marker/s for lymph node metastasis. microRNAs have been investigated as potential markers for prognosis of various types of cancer. Taking this into consideration, our main goal was to determine the association of miR-223-3p, miR-107, and miR-21-5p expression with clinicopathological characteristics, as well as presence of lymph node metastasis in patients with penile cancer. Methods: Formalin-fixed paraffin-embedded penile squamous cell carcinoma specimens from 50 patients, at diagnosis and prior to any cancer treatment, were obtained. Tissue samples comprising at least 70% malignant cells and adjacent non-tumor tissues were evaluated by using qRT-PCR for expression level of miR-223-3p, miR-107 and miR-21-5p. Additionally, molecular identification of HPV was performed by PCR, and the expression levels of PTEN were analyzed by immunohistochemistry. Results: Penile squamous cell carcinoma primary tumors presented higher expression of miR-223-3p, miR-107, and miR-21-5p when compared to non-tumor adjacent tissues. Upregulation of miR-223-3p was associated lymph node metastasis. Higher expression of miR-107 was associated with worsening of prognosis (as observed by histological grade II and III, tumors bigger than 2.0 cm, stage III and IV, and lower disease-free survival). In addition, higher expression of miR-107 and miR-21-5p was correlated to the absence of PTEN protein expression. Conclusions: Our data demonstrate that higher expression of miR-223-3p, miR-107, and miR-21-5p is correlated with poor prognosis in penile cancer. The upregulation of these microRNAs potentially affect critical cancer pathways and may be important for the prognosis and response to therapy in penile cancer.     

MiR-21/miR-375 ratio is an independent prognostic factor in patients with laryngeal squamous cell carcinoma

We sought to identify microRNAs that exhibit altered expression in laryngeal squamous cell carcinoma (SCC) and to determine whether microRNA expression is predictive of disease. This study was divided into three steps: (1) The expression of six miRNAs, such as up-regulated miR-223, miR-142-3p, miR-21, miR-16, miR-23a and down-regulated miR-375, was evaluated using total RNA isolated from freshly-frozen primary tumors and non-cancerous laryngeal squamous epithelial tissues and analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). (2) We also investigated the mRNA expression levels of processing elements (RNASEN, DGCR8, and DICER1) that participate in miRNA-biogenesis pathway. (3) We analyzed the relationships between the expression levels of these miRNAs and the clinicopathologic parameters of laryngeal SCC patients. In this study, we found that: (1) A marked difference in the microRNA expression pattern was observed between tumors and non-cancerous tissue. With regard to miRNA-processing elements, the expression level of RNASEN was higher in laryngeal SCC than in normal epithelium (P<0.01). (2) The miR-21/miR-375 expression ratio was highly sensitive and specific for disease prediction. Kaplan-Meier analysis revealed a significant association between high expression of miR-21/miR-375 in cancerous tissue and a worse prognosis (p=0.032). (3) Furthermore, the expression ratio of miR-21/mir-375 in patients with stage (III-IV) tumors was significantly higher than that in those with stage (I-II) tumors (p=0.006). These data suggest that the pattern of microRNA expression in primary laryngeal SCC tissues is exhibiting strong predictive potential.     

微小RNA-148a在非小细胞肺癌中的表达及其临床意义

目的 探讨微小RNA-148a(miR-148a)在非小细胞肺癌(NSCLC)中的表达及其与临床病理特征的关系,分析其在临床预后中的作用.方法 采用聚合酶链反应(RT-PCR)法检测48例NSCLC组织miR-148a表达水平,以miR-148a表达中位值为界,将48例患者分为两组,即高表达组和低表达组;分析miR-148a表达与患者临床病理特征的关系,采用Kaplan-Meier法估计生存率并进行单因素分析,Log-rank检验分析miR-148a表达与患者预后的关系.结果 miR-148a在NSCLC癌组织的表达量低于癌旁组织,两组差异具有统计学意义(1.052±0.659:1.397±0.667,t=2.549,P=0.013),其表达与肿瘤大小(x2=4.594,P=0.030)、淋巴结转移(x2=5.486,P=0.019)、临床分期(x2 =4.090,P=0.043)有关,而与年龄(x2=0.354,P=0.551)、性别(x2=2.277,P=0.131)、组织学类型(x2 =0.403,P=0.525)、吸烟史(x2 =0.444,P=0.505)无关.Kaplan-Meier法分析显示,miR-148a低表达患者中位生存期为23.9个月,高表达组患者中位生存期为38.7个月,差异有统计学意义(x2=4.941,P=0.026),但是中位无疾病进展生存期分别为21.6个月、29.4个月,差异无统计学意义(x2=2.168,P=0.141).结论 miR-148a在NSCLC中低表达,miR-148a低表达与NSCLC患者恶性病理特征和较差的临床预后密切相关.     

Hepatocellular carcinoma associated microRNA expression signature: integrated bioinformatics analysis,experimental validation and clinical significance

microRNA (miRNA) expression profiles varied greatly among current studies due to different technological platforms and small sample size. Systematic and integrative analysis of published datesets that compared the miRNA expression profiles between hepatocellular carcinoma (HCC) tissue and paired adjacent noncancerous liver tissue was performed to determine candidate HCC associated miRNAs. Moreover, we further validated the confirmed miRNAs in a clinical setting using qRT-PCR and Tumor Cancer Genome Atlas (TCGA) dataset. A miRNA integrated-signature of 5 upregulated and 8 downregulated miRNAs was identified from 26 published datesets in HCC using robust rank aggregation method. qRT-PCR demonstrated that miR-93-5p, miR-224-5p, miR-221-3p and miR-21-5p was increased, whereas the expression of miR-214-3p, miR-199a-3p, miR-195-5p, miR-150-5p and miR-145-5p was decreased in the HCC tissues, which was also validated on TCGA dataset. A miRNA based score using LASSO regression model provided a high accuracy for identifying HCC tissue (AUC = 0.982): HCC risk score = 0.180E_miR-221 + 0.0262E_miR-21 - 0.007E_miR-223 - 0.185E_miR-130a. E_miR-n = Log 2 (expression of microRNA n). Furthermore, expression of 5 miRNAs (miR-222, miR-221, miR-21 miR-214 and miR-130a) correlated with pathological tumor grade. Cox regression analysis showed that miR-21 was related with 3-year survival (hazard ratio [HR]: 1.509, 95%CI: 1.079–2.112, P = 0.016) and 5-year survival (HR: 1.416, 95%CI: 1.057–1.897, P = 0.020). However, none of the deregulated miRNAs was related with microscopic vascular invasion. This study provides a basis for further clinical application of miRNAs in HCC.     

miR-155-5p和ELK3在胆囊癌组织中的表达及其临床意义

目的:分析miR-155-5p和ELK3在胆囊癌组织中的表达水平,并探讨其表达与胆囊癌临床病理特征及预后的关系。方法:收集2014年1月至2015年5月在本院进行手术治疗的45例胆囊癌患者的肿瘤组织和癌旁非肿瘤组织,采用RT-PCR检测组织中miR-155-5p和ELK3 mRNA表达水平,采用免疫组织化学染色法检测组织中ELK3蛋白表达情况,分析肿瘤组织中miR-155-5p和ELK3表达水平的关系,分析miR-155-5p和ELK3表达水平与胆囊癌患者肿瘤大小、转移、浸润等病理特征及3年总生存率的关系。结果:肿瘤组织中miR-155-5p、ELK3 mRNA表达水平及ELK3蛋白阳性率均高于癌旁非肿瘤组织（P＜0.05）;肿瘤组织中miR-155-5p和ELK3表达水平呈正相关（P=0.002）;miR-155-5p表达水平与肿瘤大小、组织分化程度、淋巴结转移、远处转移、血管浸润、胆管侵犯、肝脏侵犯有关（P＜0.05）,ELK3阳性率与FIGO分期、淋巴结转移、血管浸润、胆管侵犯和肝脏侵犯有关（P＜0.05）;miR-155-5p高表达组患者3年总生存率低于miR-155-5p低表达组患者（P＜0.05）,ELK3阳性表达患者3年生存率低于ELK3阴性表达患者（P＜0.05）。结论:胆囊癌组织中miR-155-5p和ELK3表达上调,与胆囊癌转移、侵袭等恶性生物学行为及不良预后密切相关,可作为胆囊癌临床预后预测的检测指标。     

miR-182-5p在膀胱癌中的表达及其机制

目的:探究miR-182-5p及FOXO3在膀胱癌中的表达意义及其参与膀胱癌可能的机制.方法:收集2017年6月至2019年6月64例于郑州人民医院行手术切除膀胱癌患者的组织标本和临床资料,采用RT-qPCR检测膀胱癌组织及膀胱癌细胞中的miR-182-5p和FOXO3表达,按照其表达水平分为miR-182-5p高表达...