高危型HPV阳性患者血清中miR-130b-5p和miR-142-5p表达水平对宫颈癌发生的临床意义

目的 探讨血清miR-130b-5p和miR-142-5p在高危型人乳头瘤病毒(HPV)阳性的宫颈癌患者中的表达水平及其临床价值.方法 选取高危型HPV阳性宫颈癌患者76例,另选取80例高危型HPV阳性非宫颈癌患者为对照组.采用ELISA法检测2组的血清miR-130b-5p和miR-142-5p表达水平以及分析其与宫...     

前列腺癌中外周血miR-223-3p和miR-223-5p的表达水平及与临床病理特征和预后的相关性

目的探究外周血miR-223-3p和miR-223-5p水平与前列腺癌患者的临床病理特征和预后的关系。方法选取前列腺癌患者64例,RT-PCR检测患者血清miR-223-3p和miR-223-5p水平,多因素logistic回归分析前列腺癌患者预后的危险因素。结果患者年龄、术前PSA差异无统计学意义(P>0.05);血清miR-223-3p、miR-223-5p表达水平与Gleason评分、有无骨转移、TNM临床分期有关(P<0.05)。存活组与死亡组PCa患者的基线特征比较,死亡组的血清miR-223-3p、miR-223-5p水平显著高于存活组(P<0.05),Gleason评分≥7、有骨转移、TNM临床分期Ⅲ+Ⅳ期的患者存活率显著低于Gleason评分<7、无骨转移、Ⅰ+Ⅱ期患者(P<0.05)。多因素logistic回归分析显示,miR-223-3p、miR-223-5p水平上升为影响脑膜瘤患者预后的独立危险因素(P<0.05)。结论血清miR-223-3p和miR-223-5p水平与前列腺癌患者临床病理特征和预后相关。     

miR-125a-5p通过下调BAG4表达抑制胃癌细胞迁移和侵袭

目的:探讨miR-125a-5p通过调控Bcl-2相关永生基因4(Bcl-2-associated athanogene 4,BAG4)的表达抑制胃癌细胞迁移和侵袭的分子机制.方法:选用2014年1月至2015年12月兰州大学第一医院手术切除的82例胃癌组织标本及配对的癌旁组织以及人胃癌细胞系MGC803、BGC823...     

miR-93-5p和miR-106b-5p在酒精相关性肝癌中的表达及其临床意义

目的： 探讨miR-93-5p和miR-106b-5p在酒精性肝硬化肝癌（酒精相关性肝癌）患者中的表达水平及其临床意义。方法： 选取酒精相关性肝癌患者穿刺组织蜡块10例,根据性别、年龄配对原则选取10例酒精性肝硬化患者穿刺组织蜡块,采用实时荧光定量PCR（qPCR）检测组织中的miR-93-5p和miR-106b-5p表达水平。另收集河北医科大学第四医院2014年1月1日—2016年12月31日住院治疗的酒精相关性肝癌患者资料71例,治疗前抽取患者外周血,qPCR法检测血清中miR-93-5p和miR-106b-5p的表达水平,分析miR-93-5p和miR-106b-5p表达水平与患者临床病理指标和预后的关系。结果： 酒精相关性肝癌患者癌组织中miR-93-5p和miR-106b-5p的表达水平显著高于酒精性肝硬化患者（均P<0.01）。miR-93-5p或miR-106b-5p高表达组肿瘤最大径较低表达组显著增加（P<0.01）,且临床分期较晚（P<0.01）。全组患者中,年龄<60岁组患者生存率显著优于≥60岁组患者（P=0.03）;肿瘤最大径<5 cm组患者生存率显著优于≥5 cm组患者（P=0.01）;TNM分期Ⅰ+Ⅱ期患者生存率高于Ⅲ期患者（P=0.02）;miR-93-5p或miR-106b-5p高表达组患者生存率较低表达组患者显著降低（P≤0.01）;年龄和miR-106b-5p表达水平是患者预后的独立影响因素（P值分别为0.02和0.03）。结论： miR-93-5p和miR-106b-5p有作为酒精相关性肝癌肿瘤标志物的潜力,治疗前检测其表达水平可以预测患者临床病理指标和预后。     

血清miR-29a-5p和miR-222的表达与肝癌的诊断和预后的关系

目的：研究血清中miR-29a-5p和miR-222表达与肝细胞肝癌（HCC）的诊断价值和预后的关系.方法：采用实时荧光定量PCR（RT-PCR）对比检测76例HCC患者、62例肝良性疾病患者和60例健康对照组血清中miR-29a-5p和miR-222水平.分析miR-29a-5p和miR-222的表达与患者临床病理参数以及肝癌切除术预后的关系.结果：HCC患者血清miR-29a-5p和miR-222表达明显高于良性肝病和正常对照组（P＜0.05）,与AFP、肿瘤大小、癌栓、病理分化、TNM分级有关（P＜0.05）.HCC患者中miR-29a-5p和miR-222低表达组的复发转移率显著低于高表达组,其术后生存率也显著高于高表达组（P＜0.05）.结论：血清miR-29a-5p和miR-222与HCC的临床病理特征密切相关,不仅是潜在的HCC早期检测指标,且miR-29a-5p和miR-222在HCC患者血清中表达上调与HCC复发转移率高及预后差密切相关,提示其也可能是判断HCC手术预后的分子标志物.     

miR-125a-3p下调胃癌细胞NRG1表达并抑制细胞增殖

目的:探讨microRNA-125a-3p(miR-125a-3p)在胃癌细胞恶性增殖中的作用及调控机制。方法:qRT-PCR法检测各肿瘤细胞系及正常对照细胞系中miR-125a-3p的表达,应用microRNA类似物(Mimic及Inhibitor)转染技术改变细胞内miR-125a-3p表达。Western-blot检测NRG1蛋白表达水平的改变。XTT、平板克隆及裸鼠成瘤试验检测处理后胃癌细胞的增殖能力。流式细胞术检测细胞周期变化情况。结果:miR-125a-3p在多个胃癌细胞系中表达均低于永生化胃黏膜上皮细胞系(P<0.01)。利用类似物转染可以有效改变肿瘤细胞miR-125a-3p的表达(P<0.01)。miR-125a-3p过表达可以降低NRG1蛋白的表达、抑制细胞周期进展,并降低胃癌细胞系的体外和在体增殖能力,抑制miR-125a-3p表达可以见到相反结果。结论:miR-125a-3p可通过调控NRG1表达在胃癌中起到抑癌作用。miR-125a-3p可以抑制胃癌细胞的恶性增殖,因此具有成为胃癌治疗新策略候选靶点的可能。     

miR-133a-3p在胃癌组织和血浆中的表达及其对胃癌细胞增殖的影响

[摘要] 目的：检测miR-133a-3p 在胃癌组织及患者血浆中的表达水平及其对胃癌细胞增殖的影响,并探讨其与患者预后的关系。方法：收集河北医科大学第四医院普外科2012 年5 月至2013 年5 月胃癌手术切除的组织标本及术前外周静脉血标本52例,每例组织标本采集肿瘤组织(非坏死部分)和配对的癌旁组织。采用实时荧光定量RT-PCR（RT-qPCR）法检测miR-133a-3p 在胃癌组织、癌旁组织、血浆及健康体检者血浆（35 例）的表达情况,分析miR-133a-3p 的表达水平与胃癌患者中位DFS及临床病理特征的关系。CCK-8 法检测过表达或沉默miR-133a-3p 对胃癌SGC7901 细胞增殖的影响。结果：miR-133a-3p 在胃癌组织中的表达明显降低（P<0.01）,其表达水平与肿瘤TNM分期、肿瘤侵润深度（T）、淋巴结转移（N）及脉管瘤栓相关（P<0.01）;在胃癌患者血浆中的表达明显升高（P<0.01）,其表达水平与肿瘤TNM分期、淋巴结转移（N）及脉管瘤栓相关（P<0.05）;与患者血清中CA199的表达水平正相关（P<0.01）。胃癌组织中miR-133a-3p 高表达组患者中位DFS 明显高于低表达组（20.8 vs 14.8 个月,P<0.05）。血浆中miR-133a-3p 的高表达组患者中位DFS明显低于低表达组（14.4 vs 20.3 个月,P<0.05）。过表达miR-133a-3p 可明显抑制SGC7901 细胞的增殖能力,同样沉默其表达可明显促进SGC701 细胞的增殖能力（均P<0.05）。结论：miR-133a-3p 可明显抑制胃癌细胞SGC7901 的增殖能力,在胃癌组织和血浆中存在明显异常表达,其表达与患者预后明显相关,可作为胃癌早诊早治及患者临床预后判定的潜在标志物。     

miRNA-135a-5p在乳腺癌患者血清中的表达及其临床诊断效能

目的探讨miRNA-135a-5p在乳腺癌患者血清中的表达情况及其临床诊断效能。方法选取82例乳腺癌患者(乳腺癌组)、43例乳腺增生患者(乳腺增生组)和40例体检健康者(健康对照组)。分别采集3组受试者的血清样本,通过实时荧光定量聚合酶链反应(PCR)检测血清miRNA-135a-5p的相对表达量,比较不同基线特征的乳腺癌患者血清miRNA-135a-5p相对表达量,并采用受试者工作特征(ROC)曲线评价血清miRNA-135a-5p在乳腺癌中的诊断效能。结果乳腺癌组患者、乳腺增生组患者、健康对照组受试者的血清miRNA-135a-5p相对表达量分别为2.396(1.042,4.363)、0.934(0.399,1.916)、0.919(0.188,1.402);乳腺癌组患者的血清miRNA-135a-5p相对表达量高于乳腺增生组和健康对照组(P﹤0.05)。不同年龄、绝经状况、病理类型、肿瘤直径的乳腺癌患者的血清miRNA-135a-5p相对表达量比较,差异均无统计学意义(P﹥0.05);TNM分期为Ⅲ期、分化程度为低分化的乳腺癌患者的血清miRNA-135a-5p相对表达量均高于TNM分期为Ⅰ~Ⅱ期、分化程度为中高分化的患者(P﹤0.05)。采用ROC曲线评价血清miRNA-135a-5p在乳腺癌中的诊断效能,其曲线下面积为0.846(95%CI:0.608~0.972),诊断的灵敏度为81.52%,特异度为70.35%。结论与乳腺增生患者、健康体检者比较,miRNA-135a-5p在乳腺癌患者血清中的相对表达量较高,同时血清miRNA-135a-5p对乳腺癌诊断的灵敏度和特异度均较高,其可能成为乳腺癌临床诊断的生物学标志物之一。     

LncRNA TCF7在胃癌中的表达及与临床病理参数和预后的关系

目的:探讨长链非编码RNA TCF7（LncRNA TCF7）在胃癌中的表达及其与患者临床病理参数和预后的关系。方法:选取2015年2月至2016年8月本院收治的86例胃癌患者的癌组织标本为胃癌组，另选取其癌旁组织为正常组。采用实时荧光定量PCR（RT-PCR）检测胃癌组织及癌旁组织中LncRNA TCF7的相对表达量，根据检测结果中位值将其分为高表达组（57例）与低表达组（29例），观察LncRNA TCF7表达与患者临床病理参数的关系。采用Kaplan-Meier法分析LncRNA TCF7表达与患者预后的关系，并将影响胃癌患者预后的相关因素进行单因素分析及COX多因素分析。结果:胃癌组LncRNA TCF7表达水平显著高于正常组（P＜0.05）；LncRNA TCF7表达与淋巴结转移、临床分期、分化程度及浸润深度显著相关（P＜0.05）；Kaplan-Meier法分析显示LncRNA TCF7高表达组患者PFS与OS均显著低于低表达组（P＜0.05）；单因素分析显示淋巴结转移、临床分期、分化程度、浸润深度及LncRNA TCF7表达水平均为影响胃癌患者预后的危险因素；COX多因素分析显示淋巴结转移与LncRNA TCF7表达水平高低均为胃癌患者预后不良的独立危险因素。结论:LncRNA TCF7在胃癌中高表达，并与胃癌发生发展有关，同时可有效预测患者预后情况。     

Clinical and biological significance of miR-378a-3p and miR-378a-5p in colorectal cancer

To investigate miR-378a-3p and miR-378a-5p expression and their relationships with the clinicopathological features of colorectal cancer (CRC). Our results showed that miR-378a-3p and miR-378a-5p expression were dramatically lower in CRC cell lines and tissues than that in adjacent normal colorectal mucosal tissues, respectively. MiR-378a-3p and miR-378a-5p expression were significantly associated with histological differentiation and TNM stage, respectively. CRC patients with low miR-378a-3p and miR-378a-5p expression had a significantly shorter survival time than those patients with high miR-378a-3p and miR-378a-5p expression (p < 0.001, p < 0.001), respectively. Univariate and multivariable Cox regression analysis showed that tumour size, TNM stage, miR-378a-3p expression and miR-378a-5p expression were independent prognostic factors for CRC patients. Ectopic miR-378a-3p or miR-378a-5p expression inhibited cellular proliferation and colony formation, induced apoptosis and G1-phase cell cycle arrest in CRC cells, but had no effect on migration and invasion of CRC cells. Furthermore, miR-378a-3p over-expression or down-regulation could inhibit or enhance insulin-like growth factor 1 receptor (IGF1R) expression in CRC cells. There was a significantly negative correlation between IGF1R protein expression and miR-378a-3p expression in CRC tissues. MiR-378a-3p over-expression or down-regulation suppressed or enhanced phosphorylated-ERK1/2 protein level, but had no effect on phosphorylated-Akt protein level. In conclusion, miR-378a-3p and miR-378a-5p expression might play an important role as tumour suppressor gene in the initial stage of carcinogenesis of CRC.     

Clinical Significance of Upregulation of mir-196a-5p in Gastric Cancer and Enriched KEGG Pathway Analysis of Target Genes

Background: miRNAs are relatively recently discovered cancer biomarkers which have important implicationsfor cancer early diagnosis, treatment and estimation of prognosis. Here we focussed on expression of mir-196a-5pin gastric cancer tissues and cell lines so as to analyse its significance for clinicopathologic characteristics andgenerate enriched KEGG pathways clustered by target genes for exploring its potential roles as a biomarker ingastric cancer. Materials and Methods: The expression of mir-196a-5p in poorly, moderate and well differentiatedgastric cancer cell lines compared with GES-1 was detected by RT-qPCR, and the expression of mir-196a-5pin gastric cancer tissues comparing with adjacent non cancer tissues of 58 cases were also assessed by RTqPCR.Subsequently, an analysis of clinical significance of mir-196a-5p in gastric cancer and enriched KEGGpathways was executed based on the miRWalk prediction database combined with bioinformatics tools DAVID6.7 and Mirfocus 3.0. Results: RT-qPCR showed that mir-196a-5p was up-regulated in 6 poorly and moderatedifferentiated gastric cancer cell lines SGC-7901, MKN-45, MKN-28, MGC-803, BGC-823, HGC-27 comparedwith GES-1, but down-regulated in the highly differentiated gastric cancer cell line AGS. Clinical data indicatedmir-196a-5p to beup-regulated in gastric cancer tissues (47/58). Overexpression of mir-196a-5p was associatedwith more extensive degree of lymph node metastasis and clinical stage (P < 0.05; x2 test). Enriched KEGGpathway analyses of predicted and validated targets in miRWalk combined with DAVID 6.7 and Mirfocus3.0 showed that the targeted genes regulated by mir-196a-5p were involved in malignancy associated biology.Conclusions: Overexpression of mir-196a-5p is associated with lymph node metastasis and clinical stage, andenriched KEGG pathway analyses showed that targeted genes regulated by mir-196a-5p may contribute totumorgenesis, suggesting roles as an oncogenic miRNA biomarker in gastric cancer.     

Circulating exosomal miR-125a-5p as a novel biomarker for cervical cancer

Exosomal microRNAs (miRs/miRNAs) have been reported to be associated with cervical cancer. The aim of the present study was to investigate circulating exosomal miRNA as a biomarker for cervical cancer diagnosis. In the present study, samples from 6 patients with cervical cancer and 6 healthy control subjects were retrieved for exosomal RNA-sequencing. The results revealed that a total of 39 miRNAs were differentially expressed between patients with cervical cancer and healthy controls (P<0.001; fold-change >2.0). Exosomal miR-125a-5p was further quantified in plasma from 60 subjects, which included 22 healthy individuals and 38 patients with cervical cancer. miR-16a-5p served as the reference miRNA for quantitative PCR analysis of exosomal miR-125a-5p in patients with cervical cancer and healthy individuals. The results revealed that exosomal miR-125a-5p expression levels in the patients with cervical cancer were significantly lower than those in the healthy controls (P<0.001). Receiver operating characteristic (ROC) curve analyses were performed and the results revealed that the level of plasma exosomal miR-125a-5p was a potential marker for differentiating between non-cervical cancer and cervical cancer, with an ROC area under the curve of 0.7129. At the cut-off value of 2.537 for miR-125a-5p, cervical cancer diagnostic sensitivities and specificities were 59.1 and 84.2%, respectively. The present study provides confirmation that exosomal miR-125a-5p could potentially serve as a biomarker for cervical cancer diagnosis. The present study involved only a small number of clinical samples; more samples are required to support the conclusions of the present study.     

Association between miR-196a-2 Gene Polymorphism and Ovarian Cancer Prognosis in Egyptian Females

Background: Ovarian cancer is the fifth leading cause of cancer-related deaths among women worldwide. Unfortunately, early detection tests are relatively lacking. Diagnosis in the late stages of the disease carries a poor prognosis. Objective: To evaluate the relationship between miR-196a-2 rs11614913 polymorphism and ovarian cancer risk and prognosis in Egyptian females. Methods: In this case-control study, the participants were classified into 2 groups. Group A is the control group which included 50 healthy females. Group B included 50 patients newly diagnosed with ovarian carcinoma confirmed by histopathological analysis. Immunohistochemistry for P53 and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for miR-196a-2 genotypes detection were performed.   Results: There was a statistically significant difference among ovarian cancer cases and controls regarding genotypes (P = 0.003). However, the distribution of the T and C alleles in both studied groups showed no significant difference (P = 0.17). There was a statistically significant increase of CA 125 levels among CT and CC genotypes carriers of ovarian cancer cases (p = 0.04). Besides, there was a statistically significant correlation between miR-196a-2 polymorphism and each of tumor grade (P <0.001), p53 immunohistochemical expression (P= 0.002), and Figo classification (P <0.001). Conclusion: There was a statistically significant increase of CA 125 levels among C allele carriers of ovarian cancer cases. Besides, there was a statistically significant association between the miR-196a-2 polymorphism and each of tumor grade, p53 immunohistochemical expression, and Figo classification. So, miR-196a-2 polymorphism can be a possible prognostic factor in ovarian cancer.     

miR-125a-5p is a prognostic biomarker that targets HDAC4 to suppress breast tumorigenesis

Identifying stably expressed tumor markers that can be used easily to detect cancer is currently an important area of cancer research. By using miRNA microarray, we identified 20 differentially expressed miRNAs in serum samples of breast cancer patients. Expression of miR-125a-5p was relatively lower in patients with shorter survival compared to long-term survivors. In a cohort of breast cancer patients (N = 300), serum expression of miR-125a-5p was negatively and significantly correlated with tumor grade (P = 0.004), lymph-node status (P = 0.004), and tumor size (P < 0.001). Low miR-125a-5p expression was an independent prognostic marker (OR = 0.421; 95% CI = 0.184 to 0.961; P = 0.04) associated with poor survival rates (P = 0.0062). We show that miR-125a-5p directly inhibits expression of the HDAC4 gene, resulting in tumor suppression in vitro and in vivo. Together these results demonstrate that serum miR-125a-5p level in breast cancer may be a useful prognostic biomarker and offer a novel therapeutic avenue by targeting HDAC4 in breast cancer.     

Association of miR-193b Down-regulation and miR-196a up-Regulation with Clinicopathological Features and Prognosis in Gastric Cancer

Dysregulated expression of microRNAs (miRNAs) has been shown to be closely associated with tumordevelopment, progression, and carcinogenesis. However, their clinical implications for gastric cancer remainelusive. To investigate the hypothesis that genome-wide alternations of miRNAs differentiate gastric cancer tissuesfrom those matched adjacent non-tumor tissues (ANTTs), miRNA arrays were employed to examine miRNAexpression profiles for the 5-pair discovery stage, and the quantitative real-time polymerase chain reaction (qRTPCR)was applied to validate candidate miRNAs for 48-pair validation stage. Furthermore, the relationshipbetween altered miRNA and clinicopathological features and prognosis of gastric cancer was explored. Amonga total of 1,146 miRNAs analyzed, 16 miRNAs were found to be significantly different expressed in tissues fromgastric cancer compared to ANTTs (p<0.05). qRT-PCR further confirmed the variation in expression of miR-193band miR-196a in the validation stage. Down-expression of miR-193b was significantly correlated with Laurentype, differentiation, UICC stage, invasion, and metastasis of gastric cancer (p<0.05), while over-expression ofmiR-196a was significantly associated with poor differentiation (p=0.022). Moreover, binary logistic regressionanalysis demonstrated that the UICC stage was a significant risk factor for down-expression of miR-193b (adjustedOR=8.69; 95%CI=1.06-56.91; p=0.043). Additionally, Kaplan-Meier survival curves indicated that patientswith a high fold-change of down-regulated miR-193b had a significantly shorter survival time (n=19; mediansurvival=29 months) compared to patients with a low fold-change of down-regulated miR-193b (n=29; mediansurvival=54 months) (p=0.001). Overall survival time of patients with a low fold-change of up-regulated miR-196a (n=27; median survival=52 months) was significantly longer than that of patients with a high fold-changeof up-regulated miR-196a (n=21; median survival=46 months) (p=0.003). Hence, miR-193b and miR-196a maybe applied as novel and promising prognostic markers in gastric cancer.